Preferential VH gene usage in rabbit Ig-secreting heterohybridomas.
B cells from leukemic rabbits preferentially use a single VH gene, VH1, in their VDJ gene rearrangements. To determine whether Ig-secreting B cells from normal rabbits also preferentially use VH1, we generated rabbit X mouse heterohybridomas that stably secreted rabbit Ig that expressed VHa allotypic specificities and analyzed the VH genes used in their VDJ gene rearrangements. We cloned the VDJ genes from nine heterohybridomas, and by comparing the restriction map of the DNA immediately 5' of the translational start site of these VDJ genes to that of the same region 5' of VH1, we showed that eight of the nine heterohybridoma clones use VH1. Comparison of the nucleotide sequences of the eight VH1-using VDJ genes with the nucleotide sequence of germ-line VH1 showed that each of them had somatically diversified. The diversified regions included clustered nucleotide changes and codon insertions and deletions, such as would be expected if the diversification process involved somatic gene conversion. We searched our database of germ-line VH genes for genes that could serve as donors for the gene conversion events, and we identified potential VH donor genes for five regions of diversification. One of these regions of diversification spanned at least 132 bp and included a codon insertion as well as 15 nucleotide changes. The data confirm that Ab diversity may be generated by a somatic gene conversion-like mechanism. The results directly demonstrate that Ig-secreting B cells from normal rabbits preferentially use VH1 in their VDJ gene rearrangements.
Raman
C
C
Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL 60153.
Spieker-Polet
H
H
Yam
P C
PC
Knight
K L
KL
D000595:Amino Acid Sequence; D000818:Animals; D000916:Antibody Diversity; D001483:Base Sequence; D005785:Gene Conversion; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D005803:Genes, Immunoglobulin; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D015183:Restriction Mapping
D007135:Immunoglobulin Variable Region