title: The presence of autoantibodies specific for NZB serum factors in adult NZB mice and the establishment of monoclonal autoantibodies against these humoral factors.
abstract: Humoral factors in serum of young NZB mice enhance maturation of B-lymphocyte precursors in vitro. A blot ELISA assay identified autoantibodies against the serum factors. NZB-SFs (designated NZB-SF alpha, pI 3.5-4.0, and NZB-SF beta, pI 7.8) were purified by sequential steps. Both had a molecular weight (MW) of approximately 23,000 in SDS-PAGE. NZB mice develop autoantibodies against NZB-SFs by 2 months of age; titers increased progressively with age. Non-autoimmune-prone mice did not produce autoantibodies against NZB-SFs. We then developed two hybridoma clones, IIC1C1 and IIC1M4, which produce monoclonal autoantibodies against NZB-SF alpha and NZB-SF beta, respectively. Both IgM autoantibodies could be affinity purified with a column of CNBr-Sepharose 4B gel conjugated with anti-mouse IgM antibody. Neither IIC1C1 nor IIC1M4 abolished bioactivity of recombinant mouse IL-1 alpha, human IL-1, mouse, rat, or human IL-2, mouse IL-3, or colony-stimulating factor. Neither antibody reacted to recombinant mouse IL-1 alpha, IL-4, TNF alpha, or IFN gamma in blot ELISA assays. Monoclonal autoantibodies IIC1C1 and IIC1M4 were used to purify NZB-SFs. SDS-PAGE of the affinity-purified NZB-SFs revealed bands of 23 and 60 kDa, and proteins extracted from the bands were reactive to our monoclonal autoantibodies.
authors.affiliation: Department of Pediatrics, University of South Florida, St. Petersburg 33701.
authors.forename: H
authors.initials: H
authors.lastname: Jyonouchi
authors.forename: R M
authors.initials: RM
authors.lastname: Voss
authors.forename: R A
authors.initials: RA
authors.lastname: Good
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D000918:Antibody Specificity; D001323:Autoantibodies / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D003429:Cross Reactions; D005260:Female; D006825:Hybridomas / Q000032:analysis; D007163:Immunosorbent Techniques; D008222:Lymphokines / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008811:Mice, Inbred DBA; D008814:Mice, Inbred NZB / Q000276:immunology*; D008970:Molecular Weight
chemical_list: D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D008222:Lymphokines; C055837:NZB serum factor