A single VH gene is utilized predominantly in anti-BrMRBC hybridomas derived from purified Ly-1 B cells. Definition of the VH11 family

J Immunol. 1989 May 15;142(10):3643-51.

Abstract

By establishing hybridomas from two distinct surface IgM+ splenic B cell populations, Ly-1 B cells and "conventional" (Ly-1-) B cells, we found that the Ly-1 B population includes a 30 to 70 times higher frequency (1 to 2%) of cells with specificity for bromelain treated autologous red blood cells (anti-BrMRBC) when compared with conventional B cells (0.03%). We cloned and sequenced the V genes encoding anti-BrMRBC antibody from two hybridomas made with Ly-1 B cells sorted from the spleen of SM/J mice. The VH sequence (for both) is identical with the previously reported sequence associated with this specificity and belongs to a new VH gene family. This gene family, defined here as VH11, has only two members and is the predominant VH rearranged in a collection of Ly-1 B derived anti-BrMRBC hybridomas, always in association with a single VL gene (a member of the V kappa 9 family). Furthermore, analysis of hybridomas made with Ly-1 B cells sorted from the peritoneum reveals a yet higher increased frequency of VH11-encoded anti-BrMRBC specificity (30%). This variation in frequency of anti-BrMRBC in the Ly-1 population depending on location, together with the repeated association of VH11 with a particular V kappa gene suggest that antigen driven selection is (at least in part) responsible for the biased V gene expression seen in this population. Furthermore, a mechanism that might contribute to biased expression, preferential rearrangement due to close proximity to J (as seen in pre-B lines), is excluded by localization of VH11 5' to several of the more J-proximal families (Q52, 7183).

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Animals
  • Antibody Specificity
  • Antigens, Ly*
  • B-Lymphocytes / analysis*
  • B-Lymphocytes / metabolism
  • Base Sequence
  • Blood Group Antigens / genetics*
  • Blood Group Antigens / immunology
  • Bromelains
  • Cloning, Molecular
  • Female
  • Genes, Immunoglobulin*
  • Hybridomas / analysis*
  • Hybridomas / metabolism
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Heavy Chains / isolation & purification
  • Immunoglobulin Joining Region / genetics
  • Immunoglobulin Variable Region / genetics*
  • Immunoglobulin Variable Region / isolation & purification
  • Immunoglobulin kappa-Chains / genetics
  • Isoantibodies / biosynthesis
  • Isoantibodies / genetics*
  • Isoantibodies / isolation & purification
  • Mice
  • Molecular Sequence Data
  • Multigene Family
  • Phenotype
  • Polymorphism, Genetic

Substances

  • Antigens, Ly
  • Blood Group Antigens
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Joining Region
  • Immunoglobulin Variable Region
  • Immunoglobulin kappa-Chains
  • Isoantibodies
  • Bromelains