Antibodies to brain integral membrane proteins in systemic lupus erythematosus.
Wheat germ lectin affinity chromatography and temperature-induced phase separation with Triton X-114 were evaluated for the isolation of surface neuronal antigens from rat and human brain and from human neuroblastoma cell lines IMR-6 and SK-N-SH. Both techniques yielded surface proteins which were free of contamination by intracellular proteins but temperature-induced phase separation was technically less demanding and less expensive, required a shorter assay time and resulted in a superior quantity and quality of isolated proteins. Rat brain surface proteins were used for characterization of antineuronal antibody reactivity in sera from patients with systemic lupus erythematosus (SLE). Western blotting identified reactivity in 15 of 75 (20%) SLE sera compared to five of 95 (5%) normal controls (P 0.006). In rat brain the molecular weight of the individual proteins identified ranged from 59 kDa to 22 kDa. Six of these were also present in human brain and two were present in neuroblastoma cell lines. Absorption studies indicated that some of the antigenic proteins were either restricted to brain tissue or shared with other non-neuronal tissues. These techniques should facilitate the characterization of antineuronal antibody reactivities and lead to a clearer understanding of their role in the pathogenesis of autoimmune neurologic disease.
Hanly
J G
JG
Department of Medicine, Victoria General Hospital, Halifax, Nova Scotia, Canada.
Hong
C
C
D000328:Adult; D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D015153:Blotting, Western; D001921:Brain / Q000276:immunology*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley
D001323:Autoantibodies; D008565:Membrane Proteins