- Email: [email protected]
Attempted tolerization of DA rats with encephalitogenic peptide elicits experimental autoimmune encephalomyelitis (EAE) and TH1 responses
Poster Abstracts / Journal of Neuroimmunology 90 (1998) 13-105
479 Milk and Sdf Tolerance: Induction ofAutoimmune CNS Disease by a C o m m o n E n v i r o n m e n t a l A n t i g e n A. S{.efferl,Max-Plancklnstitute, Germany,M, Storch, Universityof Vienna,Austria, A. Amini, I. Mathel; UniversityofMamland, USA,H. Lassmann, Universityof Vwnna, A lzstria, C. Linington, Max-PlanckInsamte, Germany
Environmental factors play a major role in the etiology of multiple l sclerosis. In this study we report that bovine BTN, a major component of the milk fat globule membrane and therefnre a normal component of the Western diet, can trigger a pathogenic autoimmune response to MOG in the DA rat. The adoptive transfer of BTN specific T cell lines can induce extensive CNS inflammation and clinical disease. These C D 4 + T cells cross react with amino acids 71-93 u f M O G in a class II restricted fashion. In addition, extensive molecular mimicry was also observed in the B cell response to BTN by both Western blot and ELISA. These data indicate that bovine milk contains an antigen cross-reacting with the CNS component MOG at both the T and B cell level. This response is potentially pathogenic and hence may play a role in the human disease MS.
480 Characterization of Regulatory T Ceils (RTC) from Naive Lewis Rats that are Functionally Antagonistic to MBP-reaetive T Cells D. Sun, J.N. Whitaker, L. Cap, D. Wilson, UniversityofAlabama,Alabama, USA
85
482 Evidence for Gamma-delta T calls in Normal Mouse Central Nervous System B. Szvmanska. MedicalAcademyofLodz, Poland, A.J. Rajan, AlbertEinsteinCollege ofMedicine, USA,L. Battistini, tl~CCSSantaLucia, ltaly, K. Sel/n aj, MedicalAcademy ofLodz, Poland, C.F. Brosnan, AlbertEinsteinCollegeofMedicine, USA
In this study we show that a small number of gd T ceils can be detected wRhin the normal mouse central nervous system (CNS). Compared with the matching spleen cell samples, gd T cells in the CNS expressed higher levels of the T cell ectivaUon markers CD25 (IL-2 receptor a) and CD69, and lower levels of the lym phocyte homing receptor CD62L (Lselectin): SpleengdTcellsexpressedequivalentlevelsofCD45RBintandCD45RBJow whereas in the CNS only the CD45RBIow phenotypowas detected. These data indicate that gd T cells in the CNS express predominantly an activated memory/effector phenotype. Approximately 20% expressed exclusively the CDSab heterodimer, consistent with a thymic origin for these gd T cells. Compared with the total CD3+ population, spleen and CNS gd cells expressed significantly higher levels of the IL-2rb (CD122). as well as both Fes and FasL, suggesting that they may be primed to function as immunoregulato~/ T cells. RT-PCR analysis of Vd gene usage showed a predominant usage of Vd6 in CN8 gd T cells whereas most Vd genes were expressed in the spleen, Sequencing of Vd6 RTPCR demonstrated extensive junctional diversity in the spleen but a more restricted pattern within the CNS. No unique CNS sequences were found, From these data we conclude that a selective component ef the circulating gd T cell population traffics through the CNS. Thus, all major popuJetions of lymphocytes can be detected in the normal CNS and as such may play specific roles in the immunological surveillance of that organ.
483 Ast~oeyte/T Cell Interactions Affect Cytokine Secretion of Both Cell Types R. Ta_nel,D. Ram arli, M. Colam batli, Umversityof Verona,Italy, S. Goldm an, CornellMedical Cenlel; USA,B. Bonetti, G. Moretto, Universityof Verona,[taly
We have shown previously that administration of MBP-reactive T cells to naive Lewis rats induces not only EAE but also a near total resistance to subsequent disease. The disease protection paralleled with increased numbers o f a CD8 + regulatory T celI(RTC) subset and that co-i.njection of this RTC subset with encephalitogenic Tcells aborted the pathogenic activity of the latter cells. Here we show that a radio-sensitive splenic population of RTC also exists in naive rats. In co-transfer experiments, the activated RTC can neutralize the pathogenic activity of stimulatory MBP-reactive T cells in vivo. Limitinz dilution assays showed that the frequency of RTC with specificity for MBP reactive Tcells in naive rats is two orders of magnitude biglier than the frequency of MBP specific precursors, the actiwty of RTC increases substantially With age, and RTC Frequencies increase as a consec/uence of immunization with MBP reactive cells lines. Kinetic studtes have revealed a decreased ratio between RTC and MBP-reactive T cell and an increased ratio in the recovery of EAE-developing animals. Our results suggest that EAE suscept~ility may represent agapofdecreasedratio between functional RTC and autoimmune T cells, which allows a pathogenic T cell clone to escape from immuno-regulatory control.
Astrocyt.e./lymphecyle interactions are relevant for the evolution of a multiple sclemsis (MS) attack, but the mechanisms underlying and the effects generated by. this cross-talking 'in rico' are pt)orly defined. We have investigated 'in vitro" the secretion of IL-6 and IL-10 cTtokines by. human astrocytes and MBPspecific T cells when grown separetely and in conditions of co-culture. Results have shown that astrocytes secreted detectable ~imounts of IL..6. but not of ILI0; conversely. T cells secreted high levels of IL-10. but not of IL-6. In conditions of co-culture. IL-6 secretion by. astrecytes was up-regalated, together with an increased DNA binding activity of NF-kB transcription factors. Under the same conditions. IL-10 production by. T cells was significantly downregulated. Supematants from T cells did not change IL-6 production by astrocytes. These results provide evidence that the cross-talking between astrecytes and T cells induces changes in the secretorial activities of both cell types: these modifications are likely to be triggered by insoluble factors. We are currently investigating the role of several cell adhesion molecules involved in these events+
481 Attempted Tolerizalton of DA Rats with Eneephalitogenie Peptide Elicits Experimental Autoimmune Encephalomyelitis (EAE) and THI Responses
484 I m m u n o m o d u l a f i o n of E x p e r i m e n t a l A l l e r g i e Encephalomyelitis by O r a l A d m i n i s t r a t i o n of C o p o l y m e r I (Copaxone ~) D. Teitelbannt, R. Amon, M. Sela, Weizmannlnstituteof Science,Israel
D.C. Lenz, N.A. Wolf, R.B. Sm eltz, R.H. Swanborg. WayneState Universitygchool ofMedicine, USA
Tolerance to EAE can be induced in most rodents by immunization with encephalitogenic myelin basic protein (MBP) or peptide in incomplete Freund's adjuvant (IFA) prior to challenge with MBP + complete Freund's adjuvant (CFA). MBP + IFA induces immune deviation from an inflammatory THI to a protective TH2 response. In the present study, Lewis rats immunized with encephalitogenic MBP peptide + IFA failed to develop EAE. T cells expressed IL-4 mRNA but proliferated poorly to peptide. These Lewis rats are tolerant when challenged with peptide + CFA. In contrast, when we attempted to induce unresponsiveness to EAE in DA rats with encephalitogenic peptide + IFA, they developed severe EAE in response to tolerogenic treatment without further challenge. T cells proliferated vigorously to peptide and secreted high levels of the Till cytokine, IFN-y. These findings reveal important strain variations with respect to susceptibility to EAE, and suggest that DA rats have less stringent requirements for activation ofautoreactive Y cells. (Supported by NIH NS06985 and Nat. M.S. Society 1073G 10).
The activity of Copnlymcr I (Copl. Copaxone®) now knnwn as glmiramcr acetate, in suppressing e:,,perimcntal allergic cncephahmlyelills IEAE) and hi the treamlent of muhiple sclero,:is (MS) patients when injecled parenterally hits been e;~letlsiveJy delnollStla[eL[. Cop] wlts sJlown to erossleacl iulnluno}ogic~l[]y with the auloantigen myelin basic protein IMBP}. Oral administration of ataoanligcns was reported Io effectixcly stlpprcss vark~USexperin/entat amOillllllune tlisotlSCs including E A E and is n o w hcing applied to treat huTnarl ;tutoillnllUnCdb,easus like MS. W e have n()w demonstrated that oral Ct>pl iahibited E A E inducli,,m in
both rats and rnice. Disease suppression was associated with inhihitmn of both hunlora] and cellular imnlune lespoases to MBP. The secretion o1' proinfhnnnlalory cylokines was also slgnificanlly reduced. In all these assays oral Copl ,.ca>,more effecli:'e than oral MBP. The tolerance induced by oral Copl could be adop~ively tranMerred ,.,,ilh spleen cells h'om C~pl fed animals. Furthermore, Copl specific T cell lines which secrete the antJ-inflamnlatory cytokine T G F ~ and inhibil E A E hn.luclhm in vice could he isolated frlm~ lhct,c spleen celb,. In conclushm, oral Copl was showrl to h n m u m m m d u [ a t e E A E
induction iuld the hlllllUne I'esponses Io MBt:' by the induction of Th3 lype regukaory cells. These results suggesl Ihat oral tldnl[niMrgaion of Copl inay modulate MS as well.
479 Milk and Sdf Tolerance: Induction ofAutoimmune CNS Disease by a C o m m o n E n v i r o n m e n t a l A n t i g e n A. S{.efferl,Max-Plancklnstitute, Germany,M, Storch, Universityof Vienna,Austria, A. Amini, I. Mathel; UniversityofMamland, USA,H. Lassmann, Universityof Vwnna, A lzstria, C. Linington, Max-PlanckInsamte, Germany
Environmental factors play a major role in the etiology of multiple l sclerosis. In this study we report that bovine BTN, a major component of the milk fat globule membrane and therefnre a normal component of the Western diet, can trigger a pathogenic autoimmune response to MOG in the DA rat. The adoptive transfer of BTN specific T cell lines can induce extensive CNS inflammation and clinical disease. These C D 4 + T cells cross react with amino acids 71-93 u f M O G in a class II restricted fashion. In addition, extensive molecular mimicry was also observed in the B cell response to BTN by both Western blot and ELISA. These data indicate that bovine milk contains an antigen cross-reacting with the CNS component MOG at both the T and B cell level. This response is potentially pathogenic and hence may play a role in the human disease MS.
480 Characterization of Regulatory T Ceils (RTC) from Naive Lewis Rats that are Functionally Antagonistic to MBP-reaetive T Cells D. Sun, J.N. Whitaker, L. Cap, D. Wilson, UniversityofAlabama,Alabama, USA
85
482 Evidence for Gamma-delta T calls in Normal Mouse Central Nervous System B. Szvmanska. MedicalAcademyofLodz, Poland, A.J. Rajan, AlbertEinsteinCollege ofMedicine, USA,L. Battistini, tl~CCSSantaLucia, ltaly, K. Sel/n aj, MedicalAcademy ofLodz, Poland, C.F. Brosnan, AlbertEinsteinCollegeofMedicine, USA
In this study we show that a small number of gd T ceils can be detected wRhin the normal mouse central nervous system (CNS). Compared with the matching spleen cell samples, gd T cells in the CNS expressed higher levels of the T cell ectivaUon markers CD25 (IL-2 receptor a) and CD69, and lower levels of the lym phocyte homing receptor CD62L (Lselectin): SpleengdTcellsexpressedequivalentlevelsofCD45RBintandCD45RBJow whereas in the CNS only the CD45RBIow phenotypowas detected. These data indicate that gd T cells in the CNS express predominantly an activated memory/effector phenotype. Approximately 20% expressed exclusively the CDSab heterodimer, consistent with a thymic origin for these gd T cells. Compared with the total CD3+ population, spleen and CNS gd cells expressed significantly higher levels of the IL-2rb (CD122). as well as both Fes and FasL, suggesting that they may be primed to function as immunoregulato~/ T cells. RT-PCR analysis of Vd gene usage showed a predominant usage of Vd6 in CN8 gd T cells whereas most Vd genes were expressed in the spleen, Sequencing of Vd6 RTPCR demonstrated extensive junctional diversity in the spleen but a more restricted pattern within the CNS. No unique CNS sequences were found, From these data we conclude that a selective component ef the circulating gd T cell population traffics through the CNS. Thus, all major popuJetions of lymphocytes can be detected in the normal CNS and as such may play specific roles in the immunological surveillance of that organ.
483 Ast~oeyte/T Cell Interactions Affect Cytokine Secretion of Both Cell Types R. Ta_nel,D. Ram arli, M. Colam batli, Umversityof Verona,Italy, S. Goldm an, CornellMedical Cenlel; USA,B. Bonetti, G. Moretto, Universityof Verona,[taly
We have shown previously that administration of MBP-reactive T cells to naive Lewis rats induces not only EAE but also a near total resistance to subsequent disease. The disease protection paralleled with increased numbers o f a CD8 + regulatory T celI(RTC) subset and that co-i.njection of this RTC subset with encephalitogenic Tcells aborted the pathogenic activity of the latter cells. Here we show that a radio-sensitive splenic population of RTC also exists in naive rats. In co-transfer experiments, the activated RTC can neutralize the pathogenic activity of stimulatory MBP-reactive T cells in vivo. Limitinz dilution assays showed that the frequency of RTC with specificity for MBP reactive Tcells in naive rats is two orders of magnitude biglier than the frequency of MBP specific precursors, the actiwty of RTC increases substantially With age, and RTC Frequencies increase as a consec/uence of immunization with MBP reactive cells lines. Kinetic studtes have revealed a decreased ratio between RTC and MBP-reactive T cell and an increased ratio in the recovery of EAE-developing animals. Our results suggest that EAE suscept~ility may represent agapofdecreasedratio between functional RTC and autoimmune T cells, which allows a pathogenic T cell clone to escape from immuno-regulatory control.
Astrocyt.e./lymphecyle interactions are relevant for the evolution of a multiple sclemsis (MS) attack, but the mechanisms underlying and the effects generated by. this cross-talking 'in rico' are pt)orly defined. We have investigated 'in vitro" the secretion of IL-6 and IL-10 cTtokines by. human astrocytes and MBPspecific T cells when grown separetely and in conditions of co-culture. Results have shown that astrocytes secreted detectable ~imounts of IL..6. but not of ILI0; conversely. T cells secreted high levels of IL-10. but not of IL-6. In conditions of co-culture. IL-6 secretion by. astrecytes was up-regalated, together with an increased DNA binding activity of NF-kB transcription factors. Under the same conditions. IL-10 production by. T cells was significantly downregulated. Supematants from T cells did not change IL-6 production by astrocytes. These results provide evidence that the cross-talking between astrecytes and T cells induces changes in the secretorial activities of both cell types: these modifications are likely to be triggered by insoluble factors. We are currently investigating the role of several cell adhesion molecules involved in these events+
481 Attempted Tolerizalton of DA Rats with Eneephalitogenie Peptide Elicits Experimental Autoimmune Encephalomyelitis (EAE) and THI Responses
484 I m m u n o m o d u l a f i o n of E x p e r i m e n t a l A l l e r g i e Encephalomyelitis by O r a l A d m i n i s t r a t i o n of C o p o l y m e r I (Copaxone ~) D. Teitelbannt, R. Amon, M. Sela, Weizmannlnstituteof Science,Israel
D.C. Lenz, N.A. Wolf, R.B. Sm eltz, R.H. Swanborg. WayneState Universitygchool ofMedicine, USA
Tolerance to EAE can be induced in most rodents by immunization with encephalitogenic myelin basic protein (MBP) or peptide in incomplete Freund's adjuvant (IFA) prior to challenge with MBP + complete Freund's adjuvant (CFA). MBP + IFA induces immune deviation from an inflammatory THI to a protective TH2 response. In the present study, Lewis rats immunized with encephalitogenic MBP peptide + IFA failed to develop EAE. T cells expressed IL-4 mRNA but proliferated poorly to peptide. These Lewis rats are tolerant when challenged with peptide + CFA. In contrast, when we attempted to induce unresponsiveness to EAE in DA rats with encephalitogenic peptide + IFA, they developed severe EAE in response to tolerogenic treatment without further challenge. T cells proliferated vigorously to peptide and secreted high levels of the Till cytokine, IFN-y. These findings reveal important strain variations with respect to susceptibility to EAE, and suggest that DA rats have less stringent requirements for activation ofautoreactive Y cells. (Supported by NIH NS06985 and Nat. M.S. Society 1073G 10).
The activity of Copnlymcr I (Copl. Copaxone®) now knnwn as glmiramcr acetate, in suppressing e:,,perimcntal allergic cncephahmlyelills IEAE) and hi the treamlent of muhiple sclero,:is (MS) patients when injecled parenterally hits been e;~letlsiveJy delnollStla[eL[. Cop] wlts sJlown to erossleacl iulnluno}ogic~l[]y with the auloantigen myelin basic protein IMBP}. Oral administration of ataoanligcns was reported Io effectixcly stlpprcss vark~USexperin/entat amOillllllune tlisotlSCs including E A E and is n o w hcing applied to treat huTnarl ;tutoillnllUnCdb,easus like MS. W e have n()w demonstrated that oral Ct>pl iahibited E A E inducli,,m in
both rats and rnice. Disease suppression was associated with inhihitmn of both hunlora] and cellular imnlune lespoases to MBP. The secretion o1' proinfhnnnlalory cylokines was also slgnificanlly reduced. In all these assays oral Copl ,.ca>,more effecli:'e than oral MBP. The tolerance induced by oral Copl could be adop~ively tranMerred ,.,,ilh spleen cells h'om C~pl fed animals. Furthermore, Copl specific T cell lines which secrete the antJ-inflamnlatory cytokine T G F ~ and inhibil E A E hn.luclhm in vice could he isolated frlm~ lhct,c spleen celb,. In conclushm, oral Copl was showrl to h n m u m m m d u [ a t e E A E
induction iuld the hlllllUne I'esponses Io MBt:' by the induction of Th3 lype regukaory cells. These results suggesl Ihat oral tldnl[niMrgaion of Copl inay modulate MS as well.