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Multiple sclerosis: A modifying influence of HLA class I in an HLA class II associated autoimmune disease
Poster Abstracts /Journal of Neuroimmunology 90 (1998) 13-105
587 Cyclosporine A InhibitsActivation of Inducible Nitric Oxide Synthasein C6Astrocytoma
103
590
V. Traiovic, V. Badovinac, Inst. Biol. Res., Yugoslavia,V. Jankovic, M. MostaricaStojkovic, Inst.Mierob.lmmunoL Sch.Med., Yugoslavia
The effects of immunosuppresant cyclosporine A (CsA) on nitric oxide (NO) production and inducible NO synthase (iNOS) mRNA expression in C6 astrocytoma cell line were investigated. CsA (0.625-2 p_M) applied simultaneously with iNOS activator IFN-~ (200 U/ml) caused dose-dependent reduction of nitrite accumulation, an indicator of NO synthesis, in 48h culture supernatants. Proliferation of C6 cells, measured by incorporation of 3Hthymidine, was also reduced by CsA in a dose-dependent manner. However, the inhibition of NO synthesis could not be explained as merely a consequence of reduced cell growth, since it was observed also at lower concentrations of CsA (<0.5 p_M), which have not been able to affect proliferation of IFN-~'treated C6 cells. Furthermore, expression of iNOS mRNA was reduced in CsA-treated cells, indicating probably transcriptional regulation of iNOS activation by CsA. On the other hand, CsA did not have any effect on enzymatic activity of iNOS, since it failed to affect NO production in cells in which iNOS had already been induced with IFN-~' and any further induction blocked by protein synthesis inhibitor cycloheximide. Taken together, these results clearly show that CsA inhibits induction of iNOS, rather than its catalytic activity in C6 astrocytoma cell line.
588 Diversity of the Immune Reaction in Patients with Paraneoplastic Yo and Hu-antibodies: CSF Findings and Serum Western Blot Antibody Pattern U. Wurster, R. Sta~han-Kunstyr, F. Heidenreich, CSFLaboratoryMedicalSchool,
Hannover,Germany
591 Multiple Sderosis: A Modifying Influence of HLA Class I in an HLA Class II Associated Autoimmune Disease A. Foedell-Hahn, KarolinskaInstitute, Sweden,A. Ligers, HuddingeHospital, Sweden, M~ Gronning, Universityof Bergen, Sweden, J. Hillert, HuddingeHospital,Sweden, O. Olerup, KarotinskaInstitute, Sweden
In an attempt to establish additional diagnostic criteria and to broaden the knowledge about the individual immune response in paraneoplastic neurnlog~cal disease we (i) included CSF examinattons and (ii) improved the resohitiun of Western blots by employing gradient gels. 9/14 Yo-pos, patients displayed pleocytosis. 8 an increased albumin quotient, 11 an elevated IgG index and all exhibited oligoclonal bands (OB). Titration of immunostained human cerebeliar sections revealed a Yo antibody, index > 2 in 8/10. Among the 19 (90 Hu-pos. 6/16 showed pleocytnsis, 11/19 had QAIb > 7.4. in 12/19 the IgG index was raised and 17/19 had OB. The Hu antibody index was increased in 10114. In contrast to MS, CSF antibodies against measles, rubella. zosler or mumps were never seen in 19/19. Additional bands were found on Western blots of ccrebellar homogenates in the serum of 17/20 Hu-pos. patients with increased frequencies at 51-55. 60-63 and 69-73 kD. Only 5/13 Yo-pos sera showed additional bands, with one cluster at 43-46 kD. Conclusion: Any otherwise unexplainable OB pattern, espociallv of type 3 with additional identical 'bands in the serum, should lead to consider-,itiun of possible parancoplastie disease. The clinical expression may be governed both by the degree of intrathecal antibodv ~'nthesis as well as the spectrum of antibodies involved. The demunstratiun'of'antibodies with other than the standard Hu or Yo reactivities may be of pathogenetic relevance and speaks against the sole usage of recombinant antigens for diagnostic purposes.
Carrying the HLA class II haplotype DR15,DQ6 increases the risk of multiple sclerosis with 3.6. By adopting a PCR-based typing technique for HLA class I and class II genes, we have identified additional HLA class I alleles that increase and decrease the genetic susceptibility to MS. The HLA-A*0301 allele increases the risk of MS (OR=2.1) independently of DR15,DQ6. HLA-A*0201 decreases the overall risk (OR=0.52) and the presence of A*0201 reduces the risk of MS for DR15,DQ6 carriers from 3.6 to 1.8. Our findings are the first to identify a modulating effect of HLA class I alleles on the susceptibility to a human autoimmune disease; a phenomenon that has previously only been observed in animal models.
589 Common and Disparate Quantitative Trait Loci Disposing for ExperimentalArthdtis and Encephalomyelitisin the DA Rat
592 Polymorphismsin the IL-1 Gene Family are Associatedwith Age
I. Dahlm an~ J.C. Lorentzen, K.L. de Graaf, KarolinskaHospital,Sweden,A. Stefferl, C. Liningtun,Max-PIancklnstituteforPsychiatry,Germany,T. Olsson, Karolinska
Hospital, Sweden
Quantitative trait loci (QTLs) controlling inflammatory diseases with different organ specificity may hypothetically either be unique for one disease or shared among different diseases. We have investigated whether five nonMHC QTLs controlling susceptibility to experimental arthritis in DA rats, also influence myelin oligodendrocyte glycoprotein (MOG) induced experimental autoimmuue encephalomyelitis (EAE) in an DA x PVG.RTP F2 intercross. The DA allele in Cia3 (collagen induced arthritis QTL) on chromosome 4 is associated with increased severity of EAE and with high levels of anti MOG antibodies of the IgG2c subclass. Since production of antibodies oftbe IgG2c subclass may be stimulated by Thl cells, and there is evidence that such cells promote EAE, it is likely that this QTL regulate Thl cell differentiation. Furthermore, we show that the PVG allele in Oia2 (oil induced arthritis QTL) on chromosome 4 is associated with high levels of anti MOG antibodies of the IgG I subclass and of anti MOG IgE, indicating that this QTL regulate Th2 cell differentiation. These two QTLs may represent key regulators of the immune response and their further characterization are thus important for the understanding of the inflammatory process.
o f Onset i n A l z h e i m e r Disease V. Casadei, C. Fen'i, SanRaffaele ScienceInstitute, Italy, L. Caputo, Ospedale MaggiorePoliclinico,Milano,ltaly, E. Calabrese, lRCCSSantaMariaNascente,Milano, Italy, M. Franceschi, Casadi CuraS. MariaHSR Castellanza Varese,Italy, L.M.E. Grimaldi, SanRaffaeleSciencelnstinae, Italy Inflammatory mechanismsplay a role in the neurodegenerativeprocesses typicalof Alzheimer's Disease. To investigatethe genetic contributionof the interleukin-I (IL-I) cytokine familyon the occurrence and the clinical features of AD. we analyzed two distinct polymorphismsin the promoter region of IL-I alpha and IL-I beta and an intronic polymorphismof [L- I receptor antagonist (IL- Ira) in 200 sporadic AD patients and 150 ethnicallymatched healthy controls (HC). Althoughthe allele frequency (AF) of the two IL-I alpha alleles(AI and A2) did not discriminatedbetween AD patientsand tiC, there was a significantAI decrease/A2 increase (p<0.025) of AF in AD patientswith early onset (EO) compared to late onset (LO) of the type of disease. The IL-I beta AI allele was significantlyincreasedin EO-AD compared to HC (p<0.025). On the contrary, no infuence of the IL-tra potymorphismon AD was shown. In conclusion, our data seem to indicatethat IL-I genes contribute to determine the age at onset in AD. In particular the AI allele of the IL-I alpha gene is associated with a later onset ,andAI allele of the IL-I beta gene with an earlier onset in patients with AD.
587 Cyclosporine A InhibitsActivation of Inducible Nitric Oxide Synthasein C6Astrocytoma
103
590
V. Traiovic, V. Badovinac, Inst. Biol. Res., Yugoslavia,V. Jankovic, M. MostaricaStojkovic, Inst.Mierob.lmmunoL Sch.Med., Yugoslavia
The effects of immunosuppresant cyclosporine A (CsA) on nitric oxide (NO) production and inducible NO synthase (iNOS) mRNA expression in C6 astrocytoma cell line were investigated. CsA (0.625-2 p_M) applied simultaneously with iNOS activator IFN-~ (200 U/ml) caused dose-dependent reduction of nitrite accumulation, an indicator of NO synthesis, in 48h culture supernatants. Proliferation of C6 cells, measured by incorporation of 3Hthymidine, was also reduced by CsA in a dose-dependent manner. However, the inhibition of NO synthesis could not be explained as merely a consequence of reduced cell growth, since it was observed also at lower concentrations of CsA (<0.5 p_M), which have not been able to affect proliferation of IFN-~'treated C6 cells. Furthermore, expression of iNOS mRNA was reduced in CsA-treated cells, indicating probably transcriptional regulation of iNOS activation by CsA. On the other hand, CsA did not have any effect on enzymatic activity of iNOS, since it failed to affect NO production in cells in which iNOS had already been induced with IFN-~' and any further induction blocked by protein synthesis inhibitor cycloheximide. Taken together, these results clearly show that CsA inhibits induction of iNOS, rather than its catalytic activity in C6 astrocytoma cell line.
588 Diversity of the Immune Reaction in Patients with Paraneoplastic Yo and Hu-antibodies: CSF Findings and Serum Western Blot Antibody Pattern U. Wurster, R. Sta~han-Kunstyr, F. Heidenreich, CSFLaboratoryMedicalSchool,
Hannover,Germany
591 Multiple Sderosis: A Modifying Influence of HLA Class I in an HLA Class II Associated Autoimmune Disease A. Foedell-Hahn, KarolinskaInstitute, Sweden,A. Ligers, HuddingeHospital, Sweden, M~ Gronning, Universityof Bergen, Sweden, J. Hillert, HuddingeHospital,Sweden, O. Olerup, KarotinskaInstitute, Sweden
In an attempt to establish additional diagnostic criteria and to broaden the knowledge about the individual immune response in paraneoplastic neurnlog~cal disease we (i) included CSF examinattons and (ii) improved the resohitiun of Western blots by employing gradient gels. 9/14 Yo-pos, patients displayed pleocytosis. 8 an increased albumin quotient, 11 an elevated IgG index and all exhibited oligoclonal bands (OB). Titration of immunostained human cerebeliar sections revealed a Yo antibody, index > 2 in 8/10. Among the 19 (90 Hu-pos. 6/16 showed pleocytnsis, 11/19 had QAIb > 7.4. in 12/19 the IgG index was raised and 17/19 had OB. The Hu antibody index was increased in 10114. In contrast to MS, CSF antibodies against measles, rubella. zosler or mumps were never seen in 19/19. Additional bands were found on Western blots of ccrebellar homogenates in the serum of 17/20 Hu-pos. patients with increased frequencies at 51-55. 60-63 and 69-73 kD. Only 5/13 Yo-pos sera showed additional bands, with one cluster at 43-46 kD. Conclusion: Any otherwise unexplainable OB pattern, espociallv of type 3 with additional identical 'bands in the serum, should lead to consider-,itiun of possible parancoplastie disease. The clinical expression may be governed both by the degree of intrathecal antibodv ~'nthesis as well as the spectrum of antibodies involved. The demunstratiun'of'antibodies with other than the standard Hu or Yo reactivities may be of pathogenetic relevance and speaks against the sole usage of recombinant antigens for diagnostic purposes.
Carrying the HLA class II haplotype DR15,DQ6 increases the risk of multiple sclerosis with 3.6. By adopting a PCR-based typing technique for HLA class I and class II genes, we have identified additional HLA class I alleles that increase and decrease the genetic susceptibility to MS. The HLA-A*0301 allele increases the risk of MS (OR=2.1) independently of DR15,DQ6. HLA-A*0201 decreases the overall risk (OR=0.52) and the presence of A*0201 reduces the risk of MS for DR15,DQ6 carriers from 3.6 to 1.8. Our findings are the first to identify a modulating effect of HLA class I alleles on the susceptibility to a human autoimmune disease; a phenomenon that has previously only been observed in animal models.
589 Common and Disparate Quantitative Trait Loci Disposing for ExperimentalArthdtis and Encephalomyelitisin the DA Rat
592 Polymorphismsin the IL-1 Gene Family are Associatedwith Age
I. Dahlm an~ J.C. Lorentzen, K.L. de Graaf, KarolinskaHospital,Sweden,A. Stefferl, C. Liningtun,Max-PIancklnstituteforPsychiatry,Germany,T. Olsson, Karolinska
Hospital, Sweden
Quantitative trait loci (QTLs) controlling inflammatory diseases with different organ specificity may hypothetically either be unique for one disease or shared among different diseases. We have investigated whether five nonMHC QTLs controlling susceptibility to experimental arthritis in DA rats, also influence myelin oligodendrocyte glycoprotein (MOG) induced experimental autoimmuue encephalomyelitis (EAE) in an DA x PVG.RTP F2 intercross. The DA allele in Cia3 (collagen induced arthritis QTL) on chromosome 4 is associated with increased severity of EAE and with high levels of anti MOG antibodies of the IgG2c subclass. Since production of antibodies oftbe IgG2c subclass may be stimulated by Thl cells, and there is evidence that such cells promote EAE, it is likely that this QTL regulate Thl cell differentiation. Furthermore, we show that the PVG allele in Oia2 (oil induced arthritis QTL) on chromosome 4 is associated with high levels of anti MOG antibodies of the IgG I subclass and of anti MOG IgE, indicating that this QTL regulate Th2 cell differentiation. These two QTLs may represent key regulators of the immune response and their further characterization are thus important for the understanding of the inflammatory process.
o f Onset i n A l z h e i m e r Disease V. Casadei, C. Fen'i, SanRaffaele ScienceInstitute, Italy, L. Caputo, Ospedale MaggiorePoliclinico,Milano,ltaly, E. Calabrese, lRCCSSantaMariaNascente,Milano, Italy, M. Franceschi, Casadi CuraS. MariaHSR Castellanza Varese,Italy, L.M.E. Grimaldi, SanRaffaeleSciencelnstinae, Italy Inflammatory mechanismsplay a role in the neurodegenerativeprocesses typicalof Alzheimer's Disease. To investigatethe genetic contributionof the interleukin-I (IL-I) cytokine familyon the occurrence and the clinical features of AD. we analyzed two distinct polymorphismsin the promoter region of IL-I alpha and IL-I beta and an intronic polymorphismof [L- I receptor antagonist (IL- Ira) in 200 sporadic AD patients and 150 ethnicallymatched healthy controls (HC). Althoughthe allele frequency (AF) of the two IL-I alpha alleles(AI and A2) did not discriminatedbetween AD patientsand tiC, there was a significantAI decrease/A2 increase (p<0.025) of AF in AD patientswith early onset (EO) compared to late onset (LO) of the type of disease. The IL-I beta AI allele was significantlyincreasedin EO-AD compared to HC (p<0.025). On the contrary, no infuence of the IL-tra potymorphismon AD was shown. In conclusion, our data seem to indicatethat IL-I genes contribute to determine the age at onset in AD. In particular the AI allele of the IL-I alpha gene is associated with a later onset ,andAI allele of the IL-I beta gene with an earlier onset in patients with AD.