The goal of this research is to determine the genetic basis for the allotypes of the variable region of rabbit immunoglobin heavy chains. Rabbit B cell leukemias will be developed in transgenic rabbits carrying the c-myc-heavy chain enhancer gene construct. Cell lines will be developed from these rabbits and a somatic cell fusion partner will be developed for the generation of rabbit hybridomas. Rearranged VDJ genes will be cloned from the leukemic cells as well as various hybridomas in order to determine which germline genes are used in VDJ rearrangements and if there is preferential usage of one or a small number of germline V genes. These studies should provide insight into the genetic basis for the apparent allelic inheritance of rabbit VH allotypes. The genetic basis for latent VH allotypes will also be examined by preparation of a cDNA library from rabbits expressing large quantities of a latent a2 allotype. The nucleotide sequence of the cDNAs will be determined and the cDNAs will be expressed in vitro to establish which ones encode the latent a2 allotype. If a latent cDNA is cloned, the corresponding germline V gene will be cloned and analyzed by nucleotide sequence analysis and for its location within the VH chromosomal region. Finally, VHa+, and Vha- cDNA will be cloned and the nucleotide sequence determined in order to understand the structural correlates of VH allotypes. The regulatory region of genes encoding nominal VHa+, VHa- and latent allotypes will be compared in an effort to understand how the expression of the multiple VH genes is regulated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI016611-13
Application #
3126733
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1989-01-01
Project End
1992-12-31
Budget Start
1992-01-01
Budget End
1992-12-31
Support Year
13
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Loyola University Chicago
Department
Type
Schools of Medicine
DUNS #
791277940
City
Maywood
State
IL
Country
United States
Zip Code
60153
Lanning, D; Sethupathi, P; Rhee, K J et al. (2000) Intestinal microflora and diversification of the rabbit antibody repertoire. J Immunol 165:2012-9
Barrington, R A; Fasullo, M; Knight, K L (1999) A role for RAD51 in the generation of immunoglobulin gene diversity in rabbits. J Immunol 162:911-9
Winstead, C R; Zhai, S K; Sethupathi, P et al. (1999) Antigen-induced somatic diversification of rabbit IgH genes: gene conversion and point mutation. J Immunol 162:6602-12
Zhu, X; Boonthum, A; Zhai, S K et al. (1999) B lymphocyte selection and age-related changes in VH gene usage in mutant Alicia rabbits. J Immunol 163:3313-20
Vajdy, M; Sethupathi, P; Knight, K L (1998) Dependence of antibody somatic diversification on gut-associated lymphoid tissue in rabbits. J Immunol 160:2725-9
Kingzette, M; Spieker-Polet, H; Yam, P C et al. (1998) Trans-chromosomal recombination within the Ig heavy chain switch region in B lymphocytes. Proc Natl Acad Sci U S A 95:11840-5
Knight, K L; Barrington, R A (1998) Somatic diversification of IgH genes in rabbit. Immunol Rev 162:37-47
Tunyaplin, C; Knight, K L (1997) IgH gene rearrangements on the unexpressed allele in rabbit B cells. J Immunol 158:4805-11
Lanning, D K; Knight, K L (1997) Somatic hypermutation: mutations 3' of rabbit VDJ H-chain genes. J Immunol 159:4403-7
Crane, M A; Kingzette, M; Knight, K L (1996) Evidence for limited B-lymphopoiesis in adult rabbits. J Exp Med 183:2119-21

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