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Prevention of autoimmune disease by GM1-mediated modulation of lymphocyte responses
Intervention in autoimmunity
25 June 1997 - Oral presentations
diseases and 24 healthy children were also studied. Reactivity to test peptides was reootted oositive if O&&Test Peotide!Control Peptide_- .5 based on the mean iatio + 3 SD of 24 n&al child&. Resui9:Double mactivily to HBV-pal peptides and TA, NCP, NUMA. PM&l, caldesmon and my&n homologues‘w& &served in 2g%, 31%. 28%, 40%, 34% and 28%, respectively of the 65 patients with HBV infection but in none of the 88 patients with other chronic liver disease nor in healthy controls. Antibody binding to individual HBV-pol peptide was inhibited 50-100% by pre-incubation with the HBV peptide or the self homologue but not with the control peptide. Conduslon:Our results show that homologue peptides derived from HBV, nucleus and smooth muscle are targetsof antibodies in patients with chronic HBV infection. Molecular mimicry may explain the presence of ANA and SMA in chronic HBV infection.
14:00-16:00
0.5.22
0.5.22.1
Room E/F
Intervention in autoimmunity
intervention in autoimmunity with a DNA vaccine encoding the T ceil receptor p chain
Esther A.E. van Tienhoven, Jetty G. Veenstra, Willem van Eden, Chris P.M. Broeren. lnstittie of hfsctious Diseases and Immunobg~ Faculty of Veterinary Medicine, University of Utmcht, TD Utrecht, The Netherlands
introduction:Adjuvant Arthritis (AA) is an experimental autoimmune disease which can be induced by intradermal injection of Mycobacterium tuberculosis in oil. AA can be transfered by activated lymphocytes of diseased Lewis rats. An atthtitogenic T cell clone (V~ls/vcrll.3) recognizes an epitope of heat shock pmtein 85 in the context of MHC class II. In earlier experiments, immunization of rats with ovetiapping sets of peptides of VP18 allow& us to detect immunogenic T cell receptor (T&l) peptides. Moreover, the recognition of the atthritogenic T cell by TCR peptide specific T cells indicated that activated T cells can irocess and present their own TcR, thus becoming a target for regulatory anti-TCR specific T ceils. Impottantly, the natural occurrence of these cells during disease could be confirmed. The difficult task to select immunogenic peptides and the recently encouraging data on DNA vaccination experiments opened novel ways for studying the TcR anti-idiotypic responses. MaterIsIssnd Methods: The full lenght TcR j3 chain of the arthritogenic T cell clone (V,918) and the full lenght TcR of an irrelevant TcR, only differing in the vatiable domain (Va8.2) were cloned under control of a CMV promoter. 100 rg Plasmid DNA was injected intramuscular 4 times at weekly intervals 28 days prior to disease induction. Rueu)ts:The group treated with Vfll8 showed very efficient protection. This was not found in the group treated with the empty vector or in the group treated with V/38.2. The VP18 treated group still showed a response against Mycohcterium tubercukxis, indicating that there was no general immune suppression. Conclusions:Naked DNA coding for a specific TcR protects against adjuvant arthritis. This may indicate that B antior T cells recognizing V,Sl8 TcR peptides regulate the immune response. Currently. experiments are performed to further dissect the role of TCR responses in inducing protection. [O.5.22.2
371
in each group was investigated by ELISA. T-cell proliferation and cytokine production (ceil-based ELISA) were assayed in cultures of popliieal lymph nodes from the mice. Result% Injection of male DBA/l mice at the base of the tail with type II collagen in the presence of complete Freunds adjuvant normally leads to arthritis as evidenced by inflammatory infiltration and swelling of the joints. A separate injection of the GMl-binding protein, E. cdiheat-labile entemtoxin B-subunit, at the same time as collagen challenge prevented leukocyte infiltration, synovial hyperplasia and degeneration of the articular cartilage and reduced clinical symptoms of disease by 82%. The importance of GM1-interaction was demonstrated by the failure of a non-GMI-binding mutant of this protein to elicit any protective effect. Significant protection could also be achieved if administration of EtxB was delayed until 21 days after induction of disease. Analysis of T-cell responses to collagen, in cuiture.s of draining lymph node cells, revealed that protection was associated with a marked increase in IL-4 production coIKx)mitant with a reduction in ylFN levels. Furthermore, in protected mice there was a significant reduction in anti-collagen antibody levels as well as an increase in the lgGl:IgGZa ratio Conclusion: Our studies clearly indicate that GM&interaction can abrogate CIA, and suggest that agents which bind to this receptor may provide a means of preventing or treating human rheumatoid arthritis. That such protection is associated with a shift toward and anti-inflammatoty Th2 dominated response to collagen heralds the possibility that such agents will find wider therapeutic applications in controlling other inflammatory disorders. 0.5.22.3
Regulatory T ceils induced by intravenous immunogiobuiin therapy through expression of 87-2
T. Okamoto, N. Sugiura, T. Nakajima. N. Nakamura, Y. Kagitani. Department of Immunology Research Division, The Green Cross Corpotation, Osaka, Japan
intrcduction:Intravenous immunoglobulin therapy (IVIG) is an effective way of suppressing a variety of human autoimmune diseases. However, the mechanisms of action of IVIG remain unknown. Experimental autoimmune encephalomyelitis (EAE) is an autoreactive T helper type 1 (Thl) cell-mediated disease and serves as a representalive autoimmune model. To define the mechanism of action of IVIG in autoimmune disease, we examined the effect of IVIG on EAE. Materialsand Method: EAE was induced as follows. Female (PUJxSJUJ) Fl mice, aged 7-15 weeks, were immunized with rabbt myelin basic protein (MBP) or the acetylated amino terminal peptide AC 1-11 of MBP (Ac 1-11) in adjuvant complete H37Ra containing 400 pg of Killed Mycobacterium tuberculosis, strain H37Ra (Difco). Peltussis toxin (200 pg; List Biological) was given i.v. on the day of MBP immunization and 48 h later. Natural intact human immunoglobulin preparation (Venoglobulin-IH, Green Cross Corp.; 0.4 g/kg) was i.v. injected for 5 or 7 days following immunization. Results:IVIG with human immunogiobulins completely prevented the induction of EAE whether EAE was induced by either rabbit MBP or AC I-II. 87-2 was selectively induced on B cells in the draining lymph node (DLN) of the EAE-suppressed mice by IVIG both on day 3 and 8 after immunization. Treatment with anti-B7-2 mAb abrogated the inhibitory effect of IVIG in viw. On day 8 but not 3, there were many more activated T cells in the DLN of the EAE-suppressed mice by IVIG than in that of either the naive mice or the EAE-developing mice. Purified T cells from the EAEsuppressed micB by IVIG conferred passive protection against EAE to naive mice. Conclusion: These findings suggest that the selective induction of 87-2 on B cells in DLN by IVIG is critical for the induction of regulatory T cells in T cell-mediated autoimmune disease.
1 Prevention of autoimmune disease by GM-mediated modulation of lymphocyte responses
N.A. Williams, L.M. Stasuik, T.O. Nashar, C.M. Richards, A.K. Lang, M.J. Day, T.R. Hirst. Dept. of Pathology and Microbiof~ University of Bristol, UK
Introduction: Recent evidence suggests that agents which modulate the nature of the immune response may be effective in preventing or treating autoimmune disease. We recently reported that the non-toxic B-subunit of E. co/i heat-labile enterotoxin (EkB) exerts profound modulatoty effects on lymphocyte populations in vitro; notably the polyclonal activation of B-cells, apoptosis in CD8+ T-cells and to have a negligible direct effect on CD4+ T-cells. The failure of a non-receptor binding mutant of E&B, E&B (G33D), to cause such effects provided unequivocal evidence for a critical role of receptor interaction in mediating these events. The receptor which EtxB binds to is a ubiquitous cell surface glycolipid, GM1 gsnglioside. Mate&Is and Methods: Male DBA/i mice were injected with bovine lype II collagen (Cll) at the base of the tail in CFA in order to induce an autoimmune arthritis. Groups of mice were left either unprotected or were given EtxB or E&B (G33D) at a separate site either on day 0, or once disease was established, day 21. Total antibody levels and the sub-class distribution of the antiCII response
0.5.22.4
Molecular mimicry as a therapeutic approach for autoimmune disease: Oral treatment of oatients with an MHC-peptide crossreactive with’ autoantigen
S.R. Thurau, M. Diebichs-Mbhring, H. Fricke ‘, G. Wildner. ’ Saction of Immunobiology, Eye Hospital and Department of Medicine, Germany Ludwig-Maximilians-University;Munich, Germany introduction: In the rat model of experimental autoimmune uveitis we have demonstrated that a oeotide from the seauence of human disease-associated MHCclass I antigen; could induce uveitis’upon immunization. Oral administration of the MHC-peptide tolerized to the disease induced with two different retinal autoantigens, retinal S-Antigen and IRBP. The MHC-peptide cmssreacts with the major epitope from the retinal S-Antigen due to some shared discontinuous amino acid homologies. Materlais end Methods:The 14-mer peptide B27PD from the sequence of HLA-B antigens, statistically associated with uveitis, including HLA-B27, was synthesized according to GMP-condiions and encapsuled for oral application. Ten patients with long-lasting endogenous uveifs. suffering from side effects
25 June 1997 - Oral presentations
diseases and 24 healthy children were also studied. Reactivity to test peptides was reootted oositive if O&&Test Peotide!Control Peptide_- .5 based on the mean iatio + 3 SD of 24 n&al child&. Resui9:Double mactivily to HBV-pal peptides and TA, NCP, NUMA. PM&l, caldesmon and my&n homologues‘w& &served in 2g%, 31%. 28%, 40%, 34% and 28%, respectively of the 65 patients with HBV infection but in none of the 88 patients with other chronic liver disease nor in healthy controls. Antibody binding to individual HBV-pol peptide was inhibited 50-100% by pre-incubation with the HBV peptide or the self homologue but not with the control peptide. Conduslon:Our results show that homologue peptides derived from HBV, nucleus and smooth muscle are targetsof antibodies in patients with chronic HBV infection. Molecular mimicry may explain the presence of ANA and SMA in chronic HBV infection.
14:00-16:00
0.5.22
0.5.22.1
Room E/F
Intervention in autoimmunity
intervention in autoimmunity with a DNA vaccine encoding the T ceil receptor p chain
Esther A.E. van Tienhoven, Jetty G. Veenstra, Willem van Eden, Chris P.M. Broeren. lnstittie of hfsctious Diseases and Immunobg~ Faculty of Veterinary Medicine, University of Utmcht, TD Utrecht, The Netherlands
introduction:Adjuvant Arthritis (AA) is an experimental autoimmune disease which can be induced by intradermal injection of Mycobacterium tuberculosis in oil. AA can be transfered by activated lymphocytes of diseased Lewis rats. An atthtitogenic T cell clone (V~ls/vcrll.3) recognizes an epitope of heat shock pmtein 85 in the context of MHC class II. In earlier experiments, immunization of rats with ovetiapping sets of peptides of VP18 allow& us to detect immunogenic T cell receptor (T&l) peptides. Moreover, the recognition of the atthritogenic T cell by TCR peptide specific T cells indicated that activated T cells can irocess and present their own TcR, thus becoming a target for regulatory anti-TCR specific T ceils. Impottantly, the natural occurrence of these cells during disease could be confirmed. The difficult task to select immunogenic peptides and the recently encouraging data on DNA vaccination experiments opened novel ways for studying the TcR anti-idiotypic responses. MaterIsIssnd Methods: The full lenght TcR j3 chain of the arthritogenic T cell clone (V,918) and the full lenght TcR of an irrelevant TcR, only differing in the vatiable domain (Va8.2) were cloned under control of a CMV promoter. 100 rg Plasmid DNA was injected intramuscular 4 times at weekly intervals 28 days prior to disease induction. Rueu)ts:The group treated with Vfll8 showed very efficient protection. This was not found in the group treated with the empty vector or in the group treated with V/38.2. The VP18 treated group still showed a response against Mycohcterium tubercukxis, indicating that there was no general immune suppression. Conclusions:Naked DNA coding for a specific TcR protects against adjuvant arthritis. This may indicate that B antior T cells recognizing V,Sl8 TcR peptides regulate the immune response. Currently. experiments are performed to further dissect the role of TCR responses in inducing protection. [O.5.22.2
371
in each group was investigated by ELISA. T-cell proliferation and cytokine production (ceil-based ELISA) were assayed in cultures of popliieal lymph nodes from the mice. Result% Injection of male DBA/l mice at the base of the tail with type II collagen in the presence of complete Freunds adjuvant normally leads to arthritis as evidenced by inflammatory infiltration and swelling of the joints. A separate injection of the GMl-binding protein, E. cdiheat-labile entemtoxin B-subunit, at the same time as collagen challenge prevented leukocyte infiltration, synovial hyperplasia and degeneration of the articular cartilage and reduced clinical symptoms of disease by 82%. The importance of GM1-interaction was demonstrated by the failure of a non-GMI-binding mutant of this protein to elicit any protective effect. Significant protection could also be achieved if administration of EtxB was delayed until 21 days after induction of disease. Analysis of T-cell responses to collagen, in cuiture.s of draining lymph node cells, revealed that protection was associated with a marked increase in IL-4 production coIKx)mitant with a reduction in ylFN levels. Furthermore, in protected mice there was a significant reduction in anti-collagen antibody levels as well as an increase in the lgGl:IgGZa ratio Conclusion: Our studies clearly indicate that GM&interaction can abrogate CIA, and suggest that agents which bind to this receptor may provide a means of preventing or treating human rheumatoid arthritis. That such protection is associated with a shift toward and anti-inflammatoty Th2 dominated response to collagen heralds the possibility that such agents will find wider therapeutic applications in controlling other inflammatory disorders. 0.5.22.3
Regulatory T ceils induced by intravenous immunogiobuiin therapy through expression of 87-2
T. Okamoto, N. Sugiura, T. Nakajima. N. Nakamura, Y. Kagitani. Department of Immunology Research Division, The Green Cross Corpotation, Osaka, Japan
intrcduction:Intravenous immunoglobulin therapy (IVIG) is an effective way of suppressing a variety of human autoimmune diseases. However, the mechanisms of action of IVIG remain unknown. Experimental autoimmune encephalomyelitis (EAE) is an autoreactive T helper type 1 (Thl) cell-mediated disease and serves as a representalive autoimmune model. To define the mechanism of action of IVIG in autoimmune disease, we examined the effect of IVIG on EAE. Materialsand Method: EAE was induced as follows. Female (PUJxSJUJ) Fl mice, aged 7-15 weeks, were immunized with rabbt myelin basic protein (MBP) or the acetylated amino terminal peptide AC 1-11 of MBP (Ac 1-11) in adjuvant complete H37Ra containing 400 pg of Killed Mycobacterium tuberculosis, strain H37Ra (Difco). Peltussis toxin (200 pg; List Biological) was given i.v. on the day of MBP immunization and 48 h later. Natural intact human immunoglobulin preparation (Venoglobulin-IH, Green Cross Corp.; 0.4 g/kg) was i.v. injected for 5 or 7 days following immunization. Results:IVIG with human immunogiobulins completely prevented the induction of EAE whether EAE was induced by either rabbit MBP or AC I-II. 87-2 was selectively induced on B cells in the draining lymph node (DLN) of the EAE-suppressed mice by IVIG both on day 3 and 8 after immunization. Treatment with anti-B7-2 mAb abrogated the inhibitory effect of IVIG in viw. On day 8 but not 3, there were many more activated T cells in the DLN of the EAE-suppressed mice by IVIG than in that of either the naive mice or the EAE-developing mice. Purified T cells from the EAEsuppressed micB by IVIG conferred passive protection against EAE to naive mice. Conclusion: These findings suggest that the selective induction of 87-2 on B cells in DLN by IVIG is critical for the induction of regulatory T cells in T cell-mediated autoimmune disease.
1 Prevention of autoimmune disease by GM-mediated modulation of lymphocyte responses
N.A. Williams, L.M. Stasuik, T.O. Nashar, C.M. Richards, A.K. Lang, M.J. Day, T.R. Hirst. Dept. of Pathology and Microbiof~ University of Bristol, UK
Introduction: Recent evidence suggests that agents which modulate the nature of the immune response may be effective in preventing or treating autoimmune disease. We recently reported that the non-toxic B-subunit of E. co/i heat-labile enterotoxin (EkB) exerts profound modulatoty effects on lymphocyte populations in vitro; notably the polyclonal activation of B-cells, apoptosis in CD8+ T-cells and to have a negligible direct effect on CD4+ T-cells. The failure of a non-receptor binding mutant of E&B, E&B (G33D), to cause such effects provided unequivocal evidence for a critical role of receptor interaction in mediating these events. The receptor which EtxB binds to is a ubiquitous cell surface glycolipid, GM1 gsnglioside. Mate&Is and Methods: Male DBA/i mice were injected with bovine lype II collagen (Cll) at the base of the tail in CFA in order to induce an autoimmune arthritis. Groups of mice were left either unprotected or were given EtxB or E&B (G33D) at a separate site either on day 0, or once disease was established, day 21. Total antibody levels and the sub-class distribution of the antiCII response
0.5.22.4
Molecular mimicry as a therapeutic approach for autoimmune disease: Oral treatment of oatients with an MHC-peptide crossreactive with’ autoantigen
S.R. Thurau, M. Diebichs-Mbhring, H. Fricke ‘, G. Wildner. ’ Saction of Immunobiology, Eye Hospital and Department of Medicine, Germany Ludwig-Maximilians-University;Munich, Germany introduction: In the rat model of experimental autoimmune uveitis we have demonstrated that a oeotide from the seauence of human disease-associated MHCclass I antigen; could induce uveitis’upon immunization. Oral administration of the MHC-peptide tolerized to the disease induced with two different retinal autoantigens, retinal S-Antigen and IRBP. The MHC-peptide cmssreacts with the major epitope from the retinal S-Antigen due to some shared discontinuous amino acid homologies. Materlais end Methods:The 14-mer peptide B27PD from the sequence of HLA-B antigens, statistically associated with uveitis, including HLA-B27, was synthesized according to GMP-condiions and encapsuled for oral application. Ten patients with long-lasting endogenous uveifs. suffering from side effects