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Application of 2 dimensional electrophoresis (2DE) in the search for the multiple sclerosis (MS) antigen
263 serum, are available from individuals with a wide range of disorders. Because we quick freeze full coronal sections, which minimizes ice artifact formation, and photograph each section, individual structures (especially midline) can be easily identified for dissection in the frozen state and used for histochemistry. On request, the specimens can be obtained fresh, before freezing, or placed in special fixatives. Specimens are available free to investigators and are applicable to standardized biochemical, immunologic or virologic techniques. Brochures are available at the booth, or write: Dr. W.W. Tourtellotte, NNRB (W127A), VA Medical Center, Wadsworth Division, Los Angeles, CA 90073.
Application of 2 Dimensional Electrophoresis (2DE) In The Search For The Multiple Sclerosis (MS) Antigen. W.W. Tourtellotte and P. Shapshak. (VA Wadsworth Medical Center, Neurology Service, and University of California at Los Angeles, CA) It is established that intra-BBB IgG synthesis (rate of formation and unique CSF IgG oligoclonal bands) is present in almost every case of clinical definite MS (Ann Neurol 17:21, 1985). Additionally, we have established temporal invariance and clonal uniformity of lgG, A and M in MS utilizing an advanced 2-DE procedure (J Exp Med 163:41, 1986). Perhaps, the grand clue to the etiology of MS can be found if we can find the cause of the persistent intra-BBB inflammation which can be detected in vivo by its by-product intra-BBB IgG synthesis. We will present data to show that counting CSF kappa and lambda light chain clones (spots) on the final gel has an acceptable variance. Data will also be presented to show the number of spots which can be removed by absorption with measles virus of SSPE and MS prior to 2-DE.
Pertubation of the Autoimmune Idiotype Network in Experimental Autoimmune Myasthenia Gravis. J.J. Verschuuren, M.H. DeBaets, R.D. Theunissen, P. Storms, S.J. Tzartos, P.J.C. van Breda Vriesman. (Dept. of Immunology, University of Limburg, Maastricht, The Netherlands) Experimental autoimmune myasthenia gravis (EAMG) induced by injection of acetylcholine receptor (AChR) from Torpedo californica models human myasthenia gravis. We analysed the idiotype-antiidiotype network by neonatal injection of monoclonal idiotype and its complementary anti-idiotype on the immune response against AChR. Both male and female rats were injected neonatally with 50 Ilg monoclonal idiotype 65 (rat IgGl) directed at the main immunogenic region of AChR. Eleven weeks later animals were immunized with AChR (100 lag/kg) purified from the electric organ of Torpedo californica incorporated in CFA. In all animals titers against both Torpedo and rat AChR were measured by radioimmunoassay at weekly intervals. Only female rats treated neonatally with idiotype 65 had consistently elevated anti-Torpedo AChR titers from week 2 after immunisation until the end of the experiment (week 7), as compared to myeloma rat IgGl treated female animals. In the male rats no significant effect of this neonatal idiotype treatment was seen at both the anti-Torpedo and anti-rat AChR antibody titer. Next the effect of neonatal treatment with increasing doses of anti-ld 65 varying from 50 ng to 10 lag on the anti-AChR antibody titers was analysed. A high dose anti-Id (10 ~ug) suppressed anti-rat AChR titers, where as 500ng had no effect. Low dose 50 ng had a pronounced effect on the AChR antibody responses. These results demonstrate that neonatal pertubations of the idiotypeantiidiotype network profoundly alters the autoimmune response to AChR depending on the close anti-Id used suppression (pg dose) or enhancement (ng dose) of the antibody response is seen. (Supported by a grant from MEDIGON and from the Beatrix Foundation.)
Application of 2 Dimensional Electrophoresis (2DE) In The Search For The Multiple Sclerosis (MS) Antigen. W.W. Tourtellotte and P. Shapshak. (VA Wadsworth Medical Center, Neurology Service, and University of California at Los Angeles, CA) It is established that intra-BBB IgG synthesis (rate of formation and unique CSF IgG oligoclonal bands) is present in almost every case of clinical definite MS (Ann Neurol 17:21, 1985). Additionally, we have established temporal invariance and clonal uniformity of lgG, A and M in MS utilizing an advanced 2-DE procedure (J Exp Med 163:41, 1986). Perhaps, the grand clue to the etiology of MS can be found if we can find the cause of the persistent intra-BBB inflammation which can be detected in vivo by its by-product intra-BBB IgG synthesis. We will present data to show that counting CSF kappa and lambda light chain clones (spots) on the final gel has an acceptable variance. Data will also be presented to show the number of spots which can be removed by absorption with measles virus of SSPE and MS prior to 2-DE.
Pertubation of the Autoimmune Idiotype Network in Experimental Autoimmune Myasthenia Gravis. J.J. Verschuuren, M.H. DeBaets, R.D. Theunissen, P. Storms, S.J. Tzartos, P.J.C. van Breda Vriesman. (Dept. of Immunology, University of Limburg, Maastricht, The Netherlands) Experimental autoimmune myasthenia gravis (EAMG) induced by injection of acetylcholine receptor (AChR) from Torpedo californica models human myasthenia gravis. We analysed the idiotype-antiidiotype network by neonatal injection of monoclonal idiotype and its complementary anti-idiotype on the immune response against AChR. Both male and female rats were injected neonatally with 50 Ilg monoclonal idiotype 65 (rat IgGl) directed at the main immunogenic region of AChR. Eleven weeks later animals were immunized with AChR (100 lag/kg) purified from the electric organ of Torpedo californica incorporated in CFA. In all animals titers against both Torpedo and rat AChR were measured by radioimmunoassay at weekly intervals. Only female rats treated neonatally with idiotype 65 had consistently elevated anti-Torpedo AChR titers from week 2 after immunisation until the end of the experiment (week 7), as compared to myeloma rat IgGl treated female animals. In the male rats no significant effect of this neonatal idiotype treatment was seen at both the anti-Torpedo and anti-rat AChR antibody titer. Next the effect of neonatal treatment with increasing doses of anti-ld 65 varying from 50 ng to 10 lag on the anti-AChR antibody titers was analysed. A high dose anti-Id (10 ~ug) suppressed anti-rat AChR titers, where as 500ng had no effect. Low dose 50 ng had a pronounced effect on the AChR antibody responses. These results demonstrate that neonatal pertubations of the idiotypeantiidiotype network profoundly alters the autoimmune response to AChR depending on the close anti-Id used suppression (pg dose) or enhancement (ng dose) of the antibody response is seen. (Supported by a grant from MEDIGON and from the Beatrix Foundation.)