Affinity isolation of neuron-reactive antibodies in MRL/lpr mice.
Autoantibodies from the MRL/lpr mice react with numerous proteins on neuronal cell surfaces. The purpose of this study was to isolate and characterize a population of autoantibodies reactive preferentially or exclusively with nervous system tissue. Using a purified plasma membrane preparation from brain cortex of balb/c mice coupled to diaminopropylamine agarose gel, we affinity-isolated antineuronal antibodies from pooled MRL/lpr immunoglobulins. The isolated immunoglobulins reacted with brain cortex plasma membranes and neuroblastoma cells (but not liver, kidney, or fibroblasts) by Western blot and indirect immunofluorescence with confocal microscopy. By Western blot, the epitopes in the brain cortex were proteins of apparent molecular weights 101, 63, 53, 43, 39, and 33, kd; the epitopes in the neuroblastoma cells were 63, 57, and 53 kd. Lectin column isolation revealed that the 101 and 63 kd epitopes were glycosylated. Indirect immunofluorescence revealed that the antibodies bound to the cell soma more intensely than to the cell processes of viable cultured neuroblastoma cells. The cell surface localization of this binding was confirmed by confocal microscopy. Within the central nervous system the antibodies bound more intensely to primary cultures of isolated neurons from fetal cortex than to hippocampal or neostriatal cells. With these antibodies we can begin studies of their potential pathogenic effects.
Department of Neurology, Wayne State University, Detroit, Michigan.
P M
PM
Moore
I
I
Joshi
S A
SA
Ghanekar
D000818:Animals; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000302:isolation & purification*; D001327:Autoimmune Diseases / Q000276:immunology*; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D002540:Cerebral Cortex / Q000196:embryology / Q000276:immunology*; D002846:Chromatography, Affinity; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000276:immunology; D005455:Fluorescent Antibody Technique; D005782:Gels; D007163:Immunosorbent Techniques; D007668:Kidney / Q000276:immunology; D008099:Liver / Q000276:immunology; D008114:Liver Neoplasms, Experimental / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008817:Mice, Mutant Strains; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000276:immunology; D009447:Neuroblastoma / Q000276:immunology; D009474:Neurons / Q000276:immunology*; D009928:Organ Specificity; D012685:Sepharose
D001323:Autoantibodies; D005782:Gels; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D012685:Sepharose