Characterization, specificity, and IgV gene usage of anti-lymphocyte monoclonal antibodies from perinatal mice.

Academic Article

Abstract

  • 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.
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    Keywords

  • Amino Acid Sequence, Animals, Animals, Newborn, Antibodies, Monoclonal, Antibody Specificity, Base Sequence, Female, Gene Rearrangement, B-Lymphocyte, Hybridomas, Immunoglobulin M, Immunoglobulin Variable Region, Lymphocyte Subsets, Lymphoid Tissue, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Molecular Sequence Data, Sequence Alignment
  • Author List

  • Lehuen A; Bartels J; Kearney JF
  • Start Page

  • 1073
  • End Page

  • 1084
  • Volume

  • 4
  • Issue

  • 10