A single VH gene is utilized predominantly in anti-BrMRBC hybridomas derived from purified Ly-1 B cells. Definition of the VH11 family.
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).
Hardy
R R
RR
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111.
Carmack
C E
CE
Shinton
S A
SA
Riblet
R J
RJ
Hayakawa
K
K
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D000950:Antigens, Ly; D001402:B-Lymphocytes / Q000032:analysis* / Q000378:metabolism; D001483:Base Sequence; D001789:Blood Group Antigens / Q000235:genetics* / Q000276:immunology; D001963:Bromelains; D003001:Cloning, Molecular; D005260:Female; D005803:Genes, Immunoglobulin; D006825:Hybridomas / Q000032:analysis* / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000302:isolation & purification; D007133:Immunoglobulin Joining Region / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007518:Isoantibodies / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D010641:Phenotype; D011110:Polymorphism, Genetic
D000950:Antigens, Ly; D001789:Blood Group Antigens; D007143:Immunoglobulin Heavy Chains; D007133:Immunoglobulin Joining Region; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D007518:Isoantibodies; D001963:Bromelains