We use techniques of classical immunogenetics and of molecular biology to study the genetics of rabbit immunoglobulins (Igs) and T cell receptors (Tcr) and to investigate the regulated expression of Ig and Tcr genes during lymphoid cell development. We reported the thirteenth laboratory observation of a VH-CH recombination (R2M) in the rabbit. All thirteen recombinations have occurred in the male parent. The R2M recombination site maps 3'of the VH genes and 5'of the JH genes as did the sites of three previously characterized recombinations. The R2M site maps 3'of the D1d DH gene which is toward the 3'end of the DH-containing region and is probably in or near stretches of repetitive DNA. There may be two or more """"""""hot-spots of recombination"""""""" within or near stretches of repetitive DNA in the DH-containing region of the rabbit. We obtained the complete genomic sequences of the DBeta2 and JBeta2 segments associated with a chimeric CBeta gene present in some rabbits. The rabbit JBeta2 cluster has six functional segments, one pseudogene, and a remnant of another pseudogene between JBeta2.2 and JBeta2.3, equivalent to the one found in man at the same location. The JBeta2.5 gene segment of rabbit has lost the splice signal and is a pseudogene unlike its counterparts in man and mouse. The general organization of the gene segments in the DBeta2-JBeta2 regions of all three species is remarkably conserved over long stretches of DNA sequence; there is higher similarity to human than mouse. Rabbits were bred at the NIH to produce elevated levels of lambda light chains lacking c2l and expressing only c7. These rabbits were shown to produce mRNA and proteins with sequences corresponding to the products of a previously identified genomic lambda light chain gene, Clambda6. The production of c2l is known to be due to expression of Clambda5. The c2l-negative phenotype reflects deletion of a region including Jlambda5. Tbe c7-negative phenotype also appears to result from a deletion.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000036-26
Application #
3803078
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
26
Fiscal Year
1991
Total Cost
Indirect Cost
City
State
Country
United States
Zip Code
Pospisil, Richard; Alexander, Cornelius B; Obiakor, Harold et al. (2006) CD5+ B cells are preferentially expanded in rabbit appendix: the role of CD5 in B cell development and selection. Dev Comp Immunol 30:711-22
Sinha, Rajesh K; Alexander, Cornelius; Mage, Rose G (2006) Regulated expression of peripheral node addressin-positive high endothelial venules controls seeding of B lymphocytes into developing neonatal rabbit appendix. Vet Immunol Immunopathol 110:97-108
Sinha, Rajesh K; Yang, Guibin; Alexander, Cornelius et al. (2006) De novo expression of MECA-79 glycoprotein-determinant on developing B lymphocytes in gut-associated lymphoid tissues. Immunology 119:461-9
Yang, Guibin; Obiakor, Harold; Sinha, Rajesh K et al. (2005) Activation-induced deaminase cloning, localization, and protein extraction from young VH-mutant rabbit appendix. Proc Natl Acad Sci U S A 102:17083-8
Pospisil, Richard; Obiakor, Harold; Newman, Barbara A et al. (2005) Stable expression of the extracellular domains of rabbit recombinant CD5: development and characterization of polyclonal and monoclonal antibodies. Vet Immunol Immunopathol 103:257-67
Sinha, Rajesh K; Mage, Rose G (2004) Developing neonatal rabbit appendix, a primary lymphoid organ, is seeded by immature blood-borne B cells: evidence for roles for CD62L/PNAd, CCR7/CCL21, alpha4beta1 and LFA-1. Dev Comp Immunol 28:829-41
Taylor, Marcia L; Sehgal, Devinder; Raffeld, Mark et al. (2004) Demonstration that mast cells, T cells, and B cells bearing the activating kit mutation D816V occur in clusters within the marrow of patients with mastocytosis. J Mol Diagn 6:335-42
Sehgal, Devinder; Obiakor, Harold; Mage, Rose G (2002) Distinct clonal Ig diversification patterns in young appendix compared to antigen-specific splenic clones. J Immunol 168:5424-33
Obiakor, Harold; Sehgal, Devinder; Dasso, Joseph F et al. (2002) A comparison of hydraulic and laser capture microdissection methods for collection of single B cells, PCR, and sequencing of antibody VDJ. Anal Biochem 306:55-62
Sehgal, D; Schiaffella, E; Anderson, A O et al. (2000) Generation of heterogeneous rabbit anti-DNP antibodies by gene conversion and hypermutation of rearranged VL and VH genes during clonal expansion of B cells in splenic germinal centers. Eur J Immunol 30:3634-44

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