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T cell regulation within the CNS
80
Poster Abstracts / Journal of Neuroimmunology 90 (1998) 13-105
449 increased Expression of the Thl-associated Chemokinc Receptors, C X C R 3 and CCR5, by T Cells D u r i n g P r o g r e s s i v e Multiple Sclerosis K.E. Balashov, Brigham & Women'sHospital, USA,W.W. Hancock, Letd:oSite,lnc., USA, S.J. Khoury, H.L. Weiner, Brigham& Women'sHospital, USA
Objective: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) postulated to be a Thl-type T-cell mediated autoimmune disease. IFN~7, a cytokine that is the hallmark of Th 1 type immune responses, appears to play an important role in disease pathogenesis as increased production of IFN-yprecedes clinical attacks and treatment of MS patients with recombinant IFN-7 induced exacerbations of the disease. Chemokines zce essential for inflammatory responses. Recent data suggest that there is a bias in expression of selected chemokine receptors by Thl cells (CXCR3 and CCR5 positive) compared to Th2 cloned cells (CCR3 and CCR4 positive). Methodology: Peripheral blood lymphocytes were separated from both healthy control donors and different groups of MS patients. Cells were stained with antichemokine receptor specific monoclonal antibodies and analyzed by flow cyt0metry. Results and Conclusions: We detected increased number of lymphocytes that express "Thl-associated chemokine receptor" CCR5 (p < 0.01) and CXCR3 (p < 0.05) but not "Th2-associated chemokine receptor" CCR3 in progressive MS patients. CCR5 and CXCR3 receptors were expressed by both CD4+ and CD8+ T cells. We hypothesize that there is selective migration of Thi cells into the CNS in MS.
452 Androgen Treatment of Encephalitogenic T-cells A l t e r s C y t o k i n e P r o d u c t i o n a n d Reduces the Severity of E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis B.F. Bebo. Jr.. J.C. Schuster, A.A. Vandenbark, H. Offimr, OregonHealthSciences Univ.,Portland, USA
The adoptive transfer model of EAE was used to examine the role of androgens in regulating T cell cytokine secretion and the severity of disease in the SJL mouse. We recently demonstrated that PLP 139-15I specific female T cells transfer more severe EAE than male T cells and that gender differences in disease are due, at least in part, to differences in donor T ceil cytokine secretion. T cell lines were selected from PLP I39-151 immunized female SJL mice in the presence or absence of androgens. Androgen selected T cell lines secreted less IFN-y and more IL-10 than untreated cell lines. EAE induced by the transfer of androgen selected T cell lines was less severe than disease induced with untreated T cell lines. Furthermore, androgen treatment of naive TCR transgenic T cells, during their first encounter with antigen, resulted in a shift in the balance of Thl/Th2 cytokines. This phenotype was maintained during subsequent stimulations in the absence of androgen. These results suggest that androgen present in the lymphoid microenvironment during the induction of an immune response can alter the development of effector T ceils and may play a role in governing gender differences in the immune response and susceptibility to autoimmune disorders.
450
453
D e n d r i t i c Cells G e n e r a t e d in P r e s e n c e o f Intcrferon-[3 P r o m o t e T H 2
T Cell R e g u l a t i o n W i t h i n the C N S I'. Brabb, P. von Dassow, J. Goverman, Universityof Washington, Washington,USA
Responses E.J. Bartholom& L. Schanden~, P. Seldrayers, L Hildebrand, E Willems, M. Goldm an, BrasmeHospital,Belgium
Objective: To study the effect of recombinant interferon-beta (rlFN-13) on dendritic cells (DC) which are critical for the induction o f T cell responses. Methodology: Monocyte-derived DC were generated in absence or presence of 103 IU/ml rlFN-131a (IFN-[3-DC) from peripheral blood of 6 healthy volunteers. Cytokine secretion was measured using ELISA afier culture of DC with CD40-1igand-transfected fibroblasts or with allogenic T cells. Results: Spontaneous and CD40-induced p-40-1L-12 secretion (ng/ml +/SEM) was significantly lower in IFN-#-DC than in control DC (Spontaneous: 0.069 +/- 0.033 vs. 0.272 +/- 0.077, p: 0.04 and CD40-induced: 3.69 +/- 1.12 vs. 15.54 +/- 3.19, p: 0.008). Moreover, during allo-MLR, IL-5 secretion was higher when IFN-13-DC were used (0.47 +/- 0.14 vs. 0.17 +/- 0.07, p: 0.03), whereas IFN-y secretion was inhibited (3.21 +/- 0.80 vs. 9.81 +/- 5.12). Exogenous rlL-12 was able to revert 1L-5 up-regulation in alIo-MLR. Conclusion: rlFN-[3 inhibits the ability of DC to induce Thl responses and enhances the Th2 responses which are thought to be beneficial in MS.
451 In Vivo Demonstration of T Cell M i g r a t i o n a n d E l i m i n a t i o n in C N S A u t o i m m u n e Disease. Both Transgenetieally Labelled MBP-speeific and OVA-specific T Cells Undergo Apoptosis during E x p e r i m e n t a l
Mechanisms that regulate infiltration of activated T cells into the CNS and the subsequent fate of these cells is critical to understanding the development of CNS autoimmune disease. We investigated the trafficking o f T cells in the CNS in several TCR transgenic models. Our data confirms what a number of studies have previously shown, that the CNS is patrolled by a small number of activated T cells. However, our data also indicates that in the absence of activated T cells in the periphery, naive T cells can enter the CNS. The number of T cells in the CNS of OVA-specific and LCMV-specific TCR-transgenic mice is the same as in non-transgenic mice, suggesting that homeostatic mechanisms may regulate the number of T cells that enter the CNS in the absence of inflammation. The population of T cells in the CNS of MBP TCR transgenic mice is unique with regard to both the reduced number of cells and the significant down-regulation of CD44. These changes reflect regulatory mechanisms that prevent T-cell activation in the CNS in the absence of inflammatory signals.
454 N K I , I + T cells in E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis B. Cipriani, A.J. Rajan, C.E Brosnan, AlbertEinsteinCollegeofMedicine, USA, L. Battistini, IBCCS,Italy
A u t o i m m u n e Encephalomyelitis J. Bauer, Universityof Vienna,Austria, M. Bradl, MPlfor Pzychiatry,Martmsried, Germany, W.F. Hickey, DartmouthMedicalSchool, USA,C. Linington, H. Wekerle, MPIfor Psychiatry,Martinsried, Germany,H. Lassmmm, Universityof Vienna,Austria Elimination of T cells by apoptosis in the CNS of rats with experimental autoimmune encephalomyelitis (EAE) plays a role in down regulation of inflammation. We studied apoptosis in various models of EAE. Our results reveal that apoptosis of T cells is a general phenomena in EAE induced by both myelin-specific (MBP, PLP) us well as other autoimmane C N S - ~ i f i c (SI00) T cells. Elimination of T cells was present in CNS IX~renchyma but not in other CNS eomlxirtmenls like meaingus or perivascular space. We investigated whether CNS antigen- (Ag-) speeifie T cells alone or whether also CNS-irrelevant (OVAspecific) T cells ~ eliminated by apoptosis. For this we used transgenetieally (TK-tsA) labeled T cells which could he identified by ISH with a TK-tsA specific probe. This system has the advantage that the label is not lost during proliferation or apopmsis. Ag-specific T cells w¢~ seen to migrate to the CNS in large numbers during early stages (day 4) of EAE. By using either TK-tsA labeled MBP-specific or TK-IsA labeled OVA-specific T cells (co-transfea'ed with unlabeled MBP-specific T cells) we demonslmted that Ag-specitie (MBP), secondarily host-recruited as well as CNS-irrelevunt OVA-specific T lymphocyles undergoapoptosis following migration into the CNS.
The release of two cytoKines, 11_-12 and IL-4, has been shown to play a role in defining the cytokine profile of an immune response. Recently a minor subset of abTCR+ lymphocytes that in the mouse express the NKI.1 phenotype have been identified as a major source of 11_-4. A similar subset of gd TCR+ lymphocytes has also been defined. TO address the possible involvement of NKR+ T cells in experimental allergic encephalomyelitis, we have studied the distribution of these cells in the central nervous system and sptaen at varying stages in the disease process, in normal SJL mice N K I . I ÷ abTCR+ lymphecytes constituted *-0,4% of the total spleen cell population, whereas expression on gd T ceils was - 5 % . In the CNS, both the a b T C R + N K I . I + T cells and the gdTCR+ N K I , I + T cells were elevated (~2.3% and - 2 4 % respectively). In animals with EAE, no significant changes were seen in the ab or gd T C R + N K I . I + T cells in the spJeec, but in the CNS at disease onset and height the percent representation of these ceils was reduced. These data suggest that in the normal CNS, the populations of both ab T cells and gd T cells are biased towards a Th2 response, with more cells secreting IL-4. However, as the animals develop EAE the number of celts secreting IFNg increases and IL-4 decreases. As the animals recover, these numbers reverse with a greater number of ceils secreting IL-4. Thus, T cells expressing NKR show clear disease-related changes suggesting a dynamic role in the disease process.
Poster Abstracts / Journal of Neuroimmunology 90 (1998) 13-105
449 increased Expression of the Thl-associated Chemokinc Receptors, C X C R 3 and CCR5, by T Cells D u r i n g P r o g r e s s i v e Multiple Sclerosis K.E. Balashov, Brigham & Women'sHospital, USA,W.W. Hancock, Letd:oSite,lnc., USA, S.J. Khoury, H.L. Weiner, Brigham& Women'sHospital, USA
Objective: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) postulated to be a Thl-type T-cell mediated autoimmune disease. IFN~7, a cytokine that is the hallmark of Th 1 type immune responses, appears to play an important role in disease pathogenesis as increased production of IFN-yprecedes clinical attacks and treatment of MS patients with recombinant IFN-7 induced exacerbations of the disease. Chemokines zce essential for inflammatory responses. Recent data suggest that there is a bias in expression of selected chemokine receptors by Thl cells (CXCR3 and CCR5 positive) compared to Th2 cloned cells (CCR3 and CCR4 positive). Methodology: Peripheral blood lymphocytes were separated from both healthy control donors and different groups of MS patients. Cells were stained with antichemokine receptor specific monoclonal antibodies and analyzed by flow cyt0metry. Results and Conclusions: We detected increased number of lymphocytes that express "Thl-associated chemokine receptor" CCR5 (p < 0.01) and CXCR3 (p < 0.05) but not "Th2-associated chemokine receptor" CCR3 in progressive MS patients. CCR5 and CXCR3 receptors were expressed by both CD4+ and CD8+ T cells. We hypothesize that there is selective migration of Thi cells into the CNS in MS.
452 Androgen Treatment of Encephalitogenic T-cells A l t e r s C y t o k i n e P r o d u c t i o n a n d Reduces the Severity of E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis B.F. Bebo. Jr.. J.C. Schuster, A.A. Vandenbark, H. Offimr, OregonHealthSciences Univ.,Portland, USA
The adoptive transfer model of EAE was used to examine the role of androgens in regulating T cell cytokine secretion and the severity of disease in the SJL mouse. We recently demonstrated that PLP 139-15I specific female T cells transfer more severe EAE than male T cells and that gender differences in disease are due, at least in part, to differences in donor T ceil cytokine secretion. T cell lines were selected from PLP I39-151 immunized female SJL mice in the presence or absence of androgens. Androgen selected T cell lines secreted less IFN-y and more IL-10 than untreated cell lines. EAE induced by the transfer of androgen selected T cell lines was less severe than disease induced with untreated T cell lines. Furthermore, androgen treatment of naive TCR transgenic T cells, during their first encounter with antigen, resulted in a shift in the balance of Thl/Th2 cytokines. This phenotype was maintained during subsequent stimulations in the absence of androgen. These results suggest that androgen present in the lymphoid microenvironment during the induction of an immune response can alter the development of effector T ceils and may play a role in governing gender differences in the immune response and susceptibility to autoimmune disorders.
450
453
D e n d r i t i c Cells G e n e r a t e d in P r e s e n c e o f Intcrferon-[3 P r o m o t e T H 2
T Cell R e g u l a t i o n W i t h i n the C N S I'. Brabb, P. von Dassow, J. Goverman, Universityof Washington, Washington,USA
Responses E.J. Bartholom& L. Schanden~, P. Seldrayers, L Hildebrand, E Willems, M. Goldm an, BrasmeHospital,Belgium
Objective: To study the effect of recombinant interferon-beta (rlFN-13) on dendritic cells (DC) which are critical for the induction o f T cell responses. Methodology: Monocyte-derived DC were generated in absence or presence of 103 IU/ml rlFN-131a (IFN-[3-DC) from peripheral blood of 6 healthy volunteers. Cytokine secretion was measured using ELISA afier culture of DC with CD40-1igand-transfected fibroblasts or with allogenic T cells. Results: Spontaneous and CD40-induced p-40-1L-12 secretion (ng/ml +/SEM) was significantly lower in IFN-#-DC than in control DC (Spontaneous: 0.069 +/- 0.033 vs. 0.272 +/- 0.077, p: 0.04 and CD40-induced: 3.69 +/- 1.12 vs. 15.54 +/- 3.19, p: 0.008). Moreover, during allo-MLR, IL-5 secretion was higher when IFN-13-DC were used (0.47 +/- 0.14 vs. 0.17 +/- 0.07, p: 0.03), whereas IFN-y secretion was inhibited (3.21 +/- 0.80 vs. 9.81 +/- 5.12). Exogenous rlL-12 was able to revert 1L-5 up-regulation in alIo-MLR. Conclusion: rlFN-[3 inhibits the ability of DC to induce Thl responses and enhances the Th2 responses which are thought to be beneficial in MS.
451 In Vivo Demonstration of T Cell M i g r a t i o n a n d E l i m i n a t i o n in C N S A u t o i m m u n e Disease. Both Transgenetieally Labelled MBP-speeific and OVA-specific T Cells Undergo Apoptosis during E x p e r i m e n t a l
Mechanisms that regulate infiltration of activated T cells into the CNS and the subsequent fate of these cells is critical to understanding the development of CNS autoimmune disease. We investigated the trafficking o f T cells in the CNS in several TCR transgenic models. Our data confirms what a number of studies have previously shown, that the CNS is patrolled by a small number of activated T cells. However, our data also indicates that in the absence of activated T cells in the periphery, naive T cells can enter the CNS. The number of T cells in the CNS of OVA-specific and LCMV-specific TCR-transgenic mice is the same as in non-transgenic mice, suggesting that homeostatic mechanisms may regulate the number of T cells that enter the CNS in the absence of inflammation. The population of T cells in the CNS of MBP TCR transgenic mice is unique with regard to both the reduced number of cells and the significant down-regulation of CD44. These changes reflect regulatory mechanisms that prevent T-cell activation in the CNS in the absence of inflammatory signals.
454 N K I , I + T cells in E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis B. Cipriani, A.J. Rajan, C.E Brosnan, AlbertEinsteinCollegeofMedicine, USA, L. Battistini, IBCCS,Italy
A u t o i m m u n e Encephalomyelitis J. Bauer, Universityof Vienna,Austria, M. Bradl, MPlfor Pzychiatry,Martmsried, Germany, W.F. Hickey, DartmouthMedicalSchool, USA,C. Linington, H. Wekerle, MPIfor Psychiatry,Martinsried, Germany,H. Lassmmm, Universityof Vienna,Austria Elimination of T cells by apoptosis in the CNS of rats with experimental autoimmune encephalomyelitis (EAE) plays a role in down regulation of inflammation. We studied apoptosis in various models of EAE. Our results reveal that apoptosis of T cells is a general phenomena in EAE induced by both myelin-specific (MBP, PLP) us well as other autoimmane C N S - ~ i f i c (SI00) T cells. Elimination of T cells was present in CNS IX~renchyma but not in other CNS eomlxirtmenls like meaingus or perivascular space. We investigated whether CNS antigen- (Ag-) speeifie T cells alone or whether also CNS-irrelevant (OVAspecific) T cells ~ eliminated by apoptosis. For this we used transgenetieally (TK-tsA) labeled T cells which could he identified by ISH with a TK-tsA specific probe. This system has the advantage that the label is not lost during proliferation or apopmsis. Ag-specific T cells w¢~ seen to migrate to the CNS in large numbers during early stages (day 4) of EAE. By using either TK-tsA labeled MBP-specific or TK-IsA labeled OVA-specific T cells (co-transfea'ed with unlabeled MBP-specific T cells) we demonslmted that Ag-specitie (MBP), secondarily host-recruited as well as CNS-irrelevunt OVA-specific T lymphocyles undergoapoptosis following migration into the CNS.
The release of two cytoKines, 11_-12 and IL-4, has been shown to play a role in defining the cytokine profile of an immune response. Recently a minor subset of abTCR+ lymphocytes that in the mouse express the NKI.1 phenotype have been identified as a major source of 11_-4. A similar subset of gd TCR+ lymphocytes has also been defined. TO address the possible involvement of NKR+ T cells in experimental allergic encephalomyelitis, we have studied the distribution of these cells in the central nervous system and sptaen at varying stages in the disease process, in normal SJL mice N K I . I ÷ abTCR+ lymphecytes constituted *-0,4% of the total spleen cell population, whereas expression on gd T ceils was - 5 % . In the CNS, both the a b T C R + N K I . I + T cells and the gdTCR+ N K I , I + T cells were elevated (~2.3% and - 2 4 % respectively). In animals with EAE, no significant changes were seen in the ab or gd T C R + N K I . I + T cells in the spJeec, but in the CNS at disease onset and height the percent representation of these ceils was reduced. These data suggest that in the normal CNS, the populations of both ab T cells and gd T cells are biased towards a Th2 response, with more cells secreting IL-4. However, as the animals develop EAE the number of celts secreting IFNg increases and IL-4 decreases. As the animals recover, these numbers reverse with a greater number of ceils secreting IL-4. Thus, T cells expressing NKR show clear disease-related changes suggesting a dynamic role in the disease process.