Anti-fluorescein antibody 3-13 VH gene rearrangement in idiotypically cross-reactive hybridomas.
The deduced amino acid sequence of anti-fluorescein (F1) antibody 3-13 VH region (residues 1-95) was 78% homologous to the alpha-1----3-dextran binding myeloma protein J558 VH region and was in the Wu-Kabat Subgroup II or Dildrop Group I classifications. The 3-13 VH region was rearranged to a D segment with only 8 of 30 bp in common with DFL16.1 germ line D gene and less homologous to all other previously identified D sequences. This sequence was joined to the third codon of JH4. The sequence encoding VH residues 5-91 was subcloned into pSP65 and used as a probe in Southern analyses to monitor 3-13 VH gene rearrangements in 12 other anti-F1 hybridomas differentially expressing (or not at all) the 3-13 idiotype. Three clones which inhibited the 3-13 idiotype-anti-idiotype interaction as effectively as 3-13 (3-12, 3-17 and 3-35), all had rearranged a gene which hybridized to the cloned 3-13 fragment, however, each was contained on a different size restriction fragment. Analyses of five other idiotypically related (but not identical) hybridomas indicated that four had rearranged a cross hybridizing VH gene while no such rearrangements were detected among four idiotypically negative cell lines. A restriction site assay indicated five clones examined had all rearranged a Vk gene to the Jk1 or Jk2 gene segment. The sequence of the antibody 3-13 VH gene and its use in hybridization studies represent the first molecular analysis of a recurrently expressed repertoire specific idiotype within an unrestricted immune response.
Department of Microbiology, University of Illinois, Urbana 61801.
D M
DM
Reinitz
R
R
Strich
J F
JF
Scott
E W
EW
Voss
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D001483:Base Sequence; D003429:Cross Reactions; D005452:Fluoresceins; D006825:Hybridomas / Q000276:immunology*; D007130:Immunoglobulin Idiotypes / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
D000906:Antibodies; D005452:Fluoresceins; D007130:Immunoglobulin Idiotypes