Generation of the primary antibody repertoire in rabbits: expression of a diverse set of Igk-V genes may compensate for limited combinatorial diversity at the heavy chain locus |
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Authors: | Devinder Sehgal George Johnson Tai T Wu R G Mage |
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Institution: | (1) Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1892, USA e-mail: rm3z@nih.gov, Tel.: +1-301-4966113, Fax: +1-301-4960222, US;(2) Departments of Biochemistry, Molecular Biology and Cell Biology, and Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA, US |
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Abstract: | In mouse and human, generation of combinatorial diversity through use of different heavy and light chain variable region
genes in immunoglobulin rearrangements can be a major contributor to the primary antibody repertoire. In rabbits, the contribution
of the combinatorial mechanism to heavy chain diversity is minimal, as only a few Igh-V genes are rearranged and expressed. To investigate the contribution of combinatorial diversity toward generation of the rabbit
Vκ repertoire, we constructed five genomic libraries from rabbit kidney DNA and 1 cDNA library from the bone marrow of a 1-day-old
rabbit using a series of polymerase chain reaction-based strategies. Our analyses indicate that most of the sequences that
we recovered from our libraries belong to a single family and some are extremely similar. The actual number of germline Igk-V genes is potentially greater than our conservative estimate of at least 39, 28 of which we found expressed as mRNA. The germline
Igk-V genes display different lengths of the coding region 3′ of Cys 88 ranging from 7 to 12 amino acids, resulting in CDR3 length
heterogeneity among functional VκJκ sequences ranging from 8 to 15 amino acids. Some of the VκJκ junctions had N and P nucleotide additions. Thus, in contrast to limited combinatorial diversity of its heavy chain, the
rabbit can draw upon a diverse set of germline Igk-V genes. The κ light chain has the potential to be a major contributor toward generation of the antibody specificities of the
rabbit pre-immune repertoire.
Received: 14 April 1999 / Revised: 8 June 1999 |
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Keywords: | Rabbit Germline Igk-V genes Expressed Igk-V genes IGVK families Combinatorial diversity |
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