10 September 2022 University of Parma Research Repository
10-Sept-2022 Valentina Loconte1 a
Provisional list of participants
12-Dec-2014 Mr. Bruno Miguel Ponsinet Neele. Deputy Head of International. Relations ... Ms. Denis Zanotti. Official ... Paris Climat 2015.
Journal of Biological Research Journal of Journ Biological al of
Journal of Biological Research 2015; volume 88. Journal of Biological Research Correspondence: Andreina Bruno Institute of Biomedicine and Molecular.
List of participants United Nations
11-Dec-2015 Paris 30 November to 11 December 2015. List of participants. Part three ... Mr. Bruno Hoyer ... Ms. Norah Loughlin Berk. Student.
Contents
03-Jun-2017 efort president 2015/2016. Dear Participants. The Board of EFORT and the local organiser
Provisional list of participants
01-Dec-2015 Paris 30 November to 11 December 2015. Provisional list of participants ... Sr. Jorge Bruno Salgot ... Ms. Norah Loughlin Berk. Student.
Contents
efort sCientifiC Committee prague 2015 Berk Haluk ... Authors: Maia
Proxy Voting Report for January 2021 to December 2021
01-Jan-2021 Amendment to the 2015 Equity Incentive Plan. Against ... 1.3 Elect Bruno Baillavoine. Withhold ... 1.6 Elect Katherine S. Zanotti. For.
Contents
08-Jun-2015 Cáceres Palou Enric /. Berk
1 Freiberg et al: The Leipzig Catalogue of Plants (LCVP) ? An
09-May-2020 (2015):. DNA barcoding and haplotyping in different Species of ... (2015):. Evolutionary History of Blepharis (Acanthaceae) and the Origin ...
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
A6EPMNE"H .M"N6
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
8JMMJF 3UHIFS#1UMGPRE #:KGSCNOGPU LD 8 2BFTBOP 7FTP #:KGSCNOGPU LD 5 2JBOOJ 7PSBOP #:KGSCNOGPU LD 9RNGK" 0P 8BOSPPR ,% 8BMJL #/LT 2JBO 7UJHJ 8BRJPTTJOJ #:KGSCNOGPU LD .CKL<" 0P 9FVJMMF ,% 8BRSI #6RCCKOI -RUOP 8BSBMB #:KGSCNOGPU LD 86BRW -% 8UMMJS #4 2JUSFQQF 8U
REBDB #:KGSCNOGPU LD .CKL<" 0P 2JUSFQQF :BMUNCP #:KGSCNOGPU LD 3 2JBO 7UJHJ :BOBTTPOJ #:KGSCNOGPU LD 9RNGK" 0P 2JPVBOOJ :JXXUTJ #:KGSCNOGPU LD 3 8BSSJNP :RFHOPMBTP #:KGSCNOGPU LD 5 8BRL ;% ;BSFOJDL #:KGSCNOGPU LD 0IIGKLGO" :8*%
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2BFTBOP 7FTP #:KGSCNOGPU LD 5 2JBOOJ 7PSBOP #:KGSCNOGPU LD 9RNGK" 0P 8BOSPPR ,% 8BMJL #/LT 2JBO 7UJHJ 8BRJPTTJOJ #:KGSCNOGPU LD .CKL<" 0P 9FVJMMF ,% 8BRSI #6RCCKOI -RUOP 8BSBMB #:KGSCNOGPU LD 86BRW -% 8UMMJS #4 2JUSFQQF 8U
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,OHFMB 8BRJB ;JXXP #:KGSCNOGPU LD 2GI2JBDPNP ;JXXPMBTTJ #:KGSCNOGPU LD 5 ,MEP ;USTJPOJ #:KGSCNOGPU LD 3LNPF ,7WOOF .IRJSTJOF AFBVFR #:KGSCNOGPU LD ;COPCNK 4KP 8BRJP AJFSFOEBOHFR #:KGSCNOGPU LD -NG=RNE" .CNJ / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 ,,,1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 68,:0),2;
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2JBOOJ 7PSBOP #:KGSCNOGPU LD 9RNGK" 0P 8BOSPPR ,% 8BMJL #/LT 2JBO 7UJHJ 8BRJPTTJOJ #:KGSCNOGPU LD .CKL<" 0P 9FVJMMF ,% 8BRSI #6RCCKOI -RUOP 8BSBMB #:KGSCNOGPU LD 86BRW -% 8UMMJS #4 2JUSFQQF 8U
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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a ileuoe 0oissv #3A;H8DE;FI B9 04<8D=B" -F40u vuisse 2egov #3A;H8DE;FI B9 04<8D=B" -F4 Beosiue 2igoteuuv #3A;H8DE;FI B9 04<8D=B" -F4 5euose 5o Berv #3A;H8DE;FI B9 04<8D=B" -F4 2ese 8seuhoue #3A;H8DE;FI B9 04<8D=B" -F4 Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2JBO 7UJHJ 8BRJPTTJOJ #:KGSCNOGPU LD .CKL<" 0P 9FVJMMF ,% 8BRSI #6RCCKOI -RUOP 8BSBMB #:KGSCNOGPU LD 86BRW -% 8UMMJS #4 2JUSFQQF 8U
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
9FVJMMF ,% 8BRSI #6RCCKOI -RUOP 8BSBMB #:KGSCNOGPU LD 86BRW -% 8UMMJS #4 2JUSFQQF 8U
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2JUSFQQF 8U
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: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2JBO 7UJHJ :BOBTTPOJ #:KGSCNOGPU LD 9RNGK" 0P 2JPVBOOJ :JXXUTJ #:KGSCNOGPU LD 3 8BSSJNP :RFHOPMBTP #:KGSCNOGPU LD 5 8BRL ;% ;BSFOJDL #:KGSCNOGPU LD 0IIGKLGO" :8*%
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2JPVBOOJ :JXXUTJ #:KGSCNOGPU LD 3 8BSSJNP :RFHOPMBTP #:KGSCNOGPU LD 5 8BRL ;% ;BSFOJDL #:KGSCNOGPU LD 0IIGKLGO" :8*%
,OHFMB 8BRJB ;JXXP #:KGSCNOGPU LD 2GI2JBDPNP ;JXXPMBTTJ #:KGSCNOGPU LD 5 ,MEP ;USTJPOJ #:KGSCNOGPU LD 3LNPF ,7WOOF .IRJSTJOF AFBVFR #:KGSCNOGPU LD ;COPCNK 4KP 8BRJP AJFSFOEBOHFR #:KGSCNOGPU LD -NG=RNE" .CNJ / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 ,,,1
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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a ileuoe 0oissv #3A;H8DE;FI B9 04<8D=B" -F40u vuisse 2egov #3A;H8DE;FI B9 04<8D=B" -F4 Beosiue 2igoteuuv #3A;H8DE;FI B9 04<8D=B" -F4 5euose 5o Berv #3A;H8DE;FI B9 04<8D=B" -F4 2ese 8seuhoue #3A;H8DE;FI B9 04<8D=B" -F4 Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
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";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
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46F9I6/E7IG?FG0Rnh!ershu" of 4[i[]rh[2 8u[i"1
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0LB?B26F:0Rnh!ershu" of 7eno[2 8u[i"1
?GJKBFG26DE:IB0Rnh!ershu" of 4[u[nh[2 8u[i"1 F?:D616IB63BNNG0Rnh!ershu" of 9hi[n2 8u[i"1Non commercial use only0<264 ,/1 / / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 /
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 65cP
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"46-31.-2(-5/03 ,-37-)-22 :;3B6238JBegv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 9;5086R685 19776JJ33
a ileuoe 0oissv #3A;H8DE;FI B9 04<8D=B" -F40u vuisse 2egov #3A;H8DE;FI B9 04<8D=B" -F4 Beosiue 2igoteuuv #3A;H8DE;FI B9 04<8D=B" -F4 5euose 5o Berv #3A;H8DE;FI B9 04<8D=B" -F4 2ese 8seuhoue #3A;H8DE;FI B9 04<8D=B" -F4 Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
9;5086R685 19776JJ33
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: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
5euose 5o Berv #3A;H8DE;FI B9 04<8D=B" -F4 2ese 8seuhoue #3A;H8DE;FI B9 04<8D=B" -F4 Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
2ese 8seuhoue #3A;H8DE;FI B9 04<8D=B" -F4 Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
Begv :oetteugv #3A;H8DE;FI B9 04<8D=B" -F4 Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
Be!o#ov Ae :!ehoe #3A;H8DE;FI B9 04<8D=B" -F4 :ei euv Ai v #3A;H8DE;FI B9 04<8D=B" -F40u vuov Jeste #3A;H8DE;FI B9 04<8D=B" -F4 5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
5oimv Jseui e #3A;H8DE;FI B9 04<8D=B" -F4 :ov"euuo cvteissv #3A;H8DE;FI B9 04<8D=B" -F4Begissv ceoue #3A;H8DE;FI B9 04<8D=B" -F4 8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
8eugige Beoe c!ttouissv #3A;H8DE;FI B9 04<8D=B" -F4 / 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00 BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3 Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitative RT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and 1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
/ 0-;?=:29 ;5 +8;9;68329 .4>42=37 "%&(* )) &$1 / / 0<264 /1
: 3KNLI=G KC 2FKGKDFA=G 6BMB=LAE .,-/1 OKGNHB 00BV TMTZad ZN
FaZN) G8DIE AB8CC8D;E
";44,870/997870(/6/8.4,2A937471A -63?/893:A70,.4/857 ):.4ANon commercial use only Immunosuppression during spaceflight isa majorbarriertosafe lo ng-termhuman space habitation andtravel.Remarkable findingsin space have shownthat gravity changes affect important cellular mech- anisms like proliferation, differentiation,genetic expression, cytoskeletal architecture and motility inlymphocytes, monocytes and othermammalian cells.In particular, several experiments performed in space demonstrated that human T lymphocytes have remarkably reduced mitogenic activation (80-90%), thus implicating gravity as a nece ssary factor in normal immunefunction. 1,2 Subse quent space s tudiesusing soundingrockets, shuttles and International Space Station(ISS) demonstrated thatT cellactivationrequires tight con- tacts betw een each otheras well betweenTcell and monocytes as anti- gen-presenting cells. We were able toseethat cells display autonomous movements and in teractions in space. 3Moreover, we
investigatedthestructureofthecytoskeleton and in particularoftubu- lin and intermediate filamentsofvimentininJurkatcells by immunofluorescence technique on the soundingrocketMAXUS 1B. Weobserved,already 30 min after exposure tomicrogravity,a signifi- cant higher formation of large bundles of filaments, showing that the cytoskeleton undergoes important andimmediatechangesin micro- gravity. 4 Again important differences between the actin pattern between 1xg and 0xg inJ-111 cells were observed in anexperiment on boardISS. 5 Such experiments were accompanied by extensive investi- gationsperformed in the ground laboratory by the three-dimensional clinostat, called Random Positioning Machine (RPM). This machine has proven, in the last 15 years, to be a useful tool to simulate low g in the ground laboratory and to prepare space investigations.Next exper- iments conducted in sp ace and inRPMindicate that there are direct gravitational effects on the genetic expression ofIL-2 and its receptor inhumanTlumphocytes.Inour investigationontheIL-2R,wefocused our attention on the alfa and beta-chains only, because thegamma- chain is not constitutively expressed. Surprisingly,theexpression of the alfa-chain wassignificantly inhibited whereastheexpression of the b eta-chain was not influenced by microgravity. 6 Moreover,experiments in RPM usinggenearrays and quantitativeRT-PCR demonstrated that induction of 91genes was alteredin simu-lated microgravity conditions. Promoter region analysis found thatthe
majo rity ofgenesdownregulatedin microgravity were controlled by transcription factorsNFkB,CREB, ELK,AP-1 and STAT.The factthat phospho rylation of the linker of activation in T cells (LAT) is not down-regulatedin simulated microgravity indicating that cholesterol- rich lipid raftsarenot involvedin the down-regulation of the transcription factors. 7 Our LEUKIN spaceflightexperimenton board theISS allowed the evaluation of the global geneexpressionpattern of humanT cells aft er1.5 hoursofstimulationbyConA and anti- CD28 in order to iden- tify the immediateearly geneswhose transcription may be inhibited in microgravity conditions. Importantly, anonboard centrifuge was usedtogenerate a 1xg simultaneous control to isolate theeffects of microgravity from other variables of spaceflight. Microarray expres- sionanalysis after1.5 hours of activation demonstrated that 0xg and1xg- activated T cells had distinctpatterns of global gene expression
and identifi ed47genesthatwere significantlydifferentially down-reg- ula ted by at least 2 fold inmicrogravity. Expression of many genes involved in mito genesis, cytokine production, apoptosis, and signal transduction and several key immediate early genes were inhibited in microgravity. In particular, transactivation ofRel/NFkB,CREB,and SRF genetargets were down-regulated. Expression ofcREL gene tar- getswere significantly inhibited andtranscription ofcREL itselfwas signifi cantly reduced in microgravity. Analysis of gene connectivity indicated that the tumornecrosis factor(TNF) pathway is likely a maj or early downstream effector pathway inhibitedin microgravity and may lead to ineffe ctive pro-inflammatory host defenses against infec tious pathogens during spaceflight. 8 Recently, westudied the influenceofaltered gravityon expres- sion and fun ction of cytoskeletal proteins,chemokines,cytokines and theirrceptors by theexperiment STIM (Signal TransductionIn Microgravity) on board the soundingrocket Maser 12. The launch took placethe 13 th offeruary 2012atEsrange Space Center and themicro- gravity lasted 390sec.Duringthe flight, one automedplunger activa- tion mechanisminitiated the confluente between the activators (Concanavalin A, anti-CD28, anti-CD3) and the cells (human Tlym- pho cytes), while a second plungerinitiated thatbetween fixative (for- malin) and activated cells inasubsequent phase. The hypergravity phase during thelaunch resultedin a down regulation of the IL-2 and CD3receptor and reduction oftyrosine phosphorylation, p44/42-MAPK phospho rylation and histone H3 acetylation,whereas LATphosphory- lationwas increased. Comparedto the baseline situationatthe point of entryinto the microgravityphase,CD3 and IL-2 receptor expression at the surfaceof non-activated T cells were reduced after 6 min. of microgravity. Importantly,p44/42-MAPK- phosphorylation was also reduc edin lowgravity.In activated T cells, the reducedCD3 andIL-2quotesdbs_dbs25.pdfusesText_31
[PDF] berck ville berck plage - Office de Tourisme de Berck-sur-Mer - France
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[PDF] Berck-Bellevue et ligne Berck-Plage - Paris-Plage - Gestion De Projet
[PDF] Berck-sur-mer, Musée Opale Sud Le Musée Les collections
[PDF] Berck-sur-Mer, un véritable village vacances durant l`été - Anciens Et Réunions
[PDF] bercofer primer st
[PDF] BERDAH DAVID Né le 20 mai 1985 à Marseille (13) Doctorant en
[PDF] Berec Immobilier Berec Immobilier
[PDF] Berechit Au commencement - El - Saints Des Derniers Jours
[PDF] Berechnung der Verzugszinsen Bauhandwerk: Vereinbarung der VOB
[PDF] Berechnung des Korrekturfaktors für
[PDF] Berechnung des Kupferzuschlages
[PDF] Berechnung von Hebeanlagen und Pumpstationen
[PDF] Berechnungsbeispiel Dehler 32 - Anciens Et Réunions
[PDF] Berechnungsbogen für die Beratungshilfe und Prozesskostenhilfe
[PDF] Berechnungsmatrix Bildungsscheck 500 € (Stand: 12.05.2015)
