Characterization, specificity, and IgV gene usage of anti-lymphocyte monoclonal antibodies from perinatal mice.
Previous studies have shown that CD5+ B cells predominate during development of the immune system and frequently secrete self-reactive antibodies, some of which appear to influence the development of the adult B cell repertoire. In addition, we now show that a high frequency of perinatally derived antibodies react with lymphocytes. Hybridomas derived from perinatal liver and splenic B cells and from spleens of adult BALB/c and C57BL/6 mice were screened by immunofluorescence on thymocytes. Anti-lymphocyte antibodies, all of the IgM isotype, were detected at a high frequency from perinatal fusions, but none were obtained from adult mice. These anti-lymphocyte mAbs were heterogeneous because they stained different subsets of peripheral T and B lymphocytes. Although the antigens recognized by these mAbs were heterogeneous with respect to their sensitivity to a variety of enzymes, 13 of the 19 mAbs recognized epitopes which were modulated by phosphatidylinositol-phospholipase C treatment. Inhibition experiments suggested that six of these 13 mAbs shared the same molecular specificity, and that they recognized the same T cell subset (62% of CD4+ and 98% of CD8+ cells). Furthermore, three of these mAbs immunoprecipitated the same 100 kDa protein from thymocytes (70 kDa in reducing conditions). The related molecular specificity of some anti-lymphocyte mAbs was also reflected by their restricted V gene usage. Three of the five mAbs specific for the 100 kDa protein used very similar or identical germline SM7 VH genes. In addition to using the same germline D and JH genes, they also exhibited identical VH-D-JH joins, despite originating from distinct fusions. Analysis of light chains also showed some restriction by preferential use of germline V kappa 4 and J kappa 5 genes. Together, these results suggest that the restricted antibody repertoire characteristic of mouse fetal and neonatal B cells is also reflected in the production of anti-lymphocyte antibodies. These B cells appear consistently in early development, use germline V genes, and express a characteristic VH-D-JH join.
Lehuen
A
A
Department of Microbiology, University of Alabama, Birmingham 35294.
Bartels
J
J
Kearney
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D001483:Base Sequence; D005260:Female; D015322:Gene Rearrangement, B-Lymphocyte; D006825:Hybridomas / Q000276:immunology; D007075:Immunoglobulin M / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D016131:Lymphocyte Subsets / Q000276:immunology*; D008221:Lymphoid Tissue / Q000196:embryology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C / Q000196:embryology / Q000235:genetics / Q000276:immunology*; D008810:Mice, Inbred C57BL / Q000196:embryology / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region