Frequency analysis of functional immunoglobulin C and V gene expression in murine B cells at various ages

J Immunol. 1983 Oct;131(4):1629-34.

Abstract

The frequencies of lipopolysaccharide- (LPS) reactive B cells and their antibody specificity repertoire have been determined in the spleen and bone marrow (BM) of mice at different ages. A limiting dilution culture system was employed that allows the growth and development of every LPS-reactive B cell into a clone an IgM-secreting cells that are capable of switching to other Ig heavy chain isotypes (C gene expression). The secretion of IgM and IgG1 was assessed in the protein A plaque assay, whereas specific IgM antibody-secreting cells (V gene expression) were detected with the use of plaque assays specific for various heterologous erythrocytes and sheep red blood cells (SRBC) coupled with a number of different haptens. The frequencies of LPS-reactive B cells in the spleen and BM of C3H/Tif, C57BL/Ka, BALB/c, and CBA/Rij mice appeared to be similar in 6- to 12- and 100-wk-old animals, as was the switch frequency to IgG1 secretion in three strains tested. Moreover, no age-related changes were observed in the frequencies of antigen-specific B cells within the pool of LPS-reactive B cells in the spleen and BM of C57BL/Ka mice. The frequencies ranged from 1 in 10 to 1 in 20 for NIP4- and NNP2-SRBC, from 1 in 50 to 1 in 100 for TNP30-SRBC, and from 1 in 1000 to 1 in 4000 for SRBC, HRBC, and GRBC. The specificity repertoire of the "spontaneously" occurring ("background") IgM-secreting cells in the spleen and BM, on the other hand, did differ between young and old C57BL/Ka mice. During aging the frequencies of the tested specificities decreased in the spleen but increased in the BM. Our data indicate that in unintentionally immunized mice the clonal selection of B lineage cells by antigen takes place at the level of the mature, antigen-reactive B cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging*
  • Animals
  • Antibody Specificity
  • Antibody-Producing Cells / immunology
  • Antibody-Producing Cells / physiology
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / physiology
  • Bone Marrow Cells
  • Cell Count
  • Female
  • Immunoglobulin Constant Regions / genetics*
  • Immunoglobulin Constant Regions / immunology
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin M / biosynthesis
  • Immunoglobulin Variable Region / genetics*
  • Immunoglobulin Variable Region / immunology
  • Immunoglobulins / genetics*
  • Lipopolysaccharides / pharmacology
  • Lymphocyte Activation
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Rats
  • Rats, Inbred Lew
  • Spleen / cytology

Substances

  • Immunoglobulin Constant Regions
  • Immunoglobulin G
  • Immunoglobulin M
  • Immunoglobulin Variable Region
  • Immunoglobulins
  • Lipopolysaccharides