The general objective of the proposed research is to establish molecular mechanisms for the control of immunoglobulin (Ig) gene expression in human normal and disease states. Specifically, the organization and regulation of expression of the human lambda (Lambda) light chain V (variable), J (joining) and C (constant) gene segments will be studied. Specific objectives include the localization, isolation and characterization of JLambda gene segments, isolation of DNA probes which will detect specific VLambda subgroups and Clambda isotypes, and cloning and sequencing germline and rearranged (active) Vlambda gene segments. This project will give information on the molecular organization of the human Lambda genes, the degree and location of somatic diversification in the VLambda gene segments, which JLambda and CLambda gene segments are functional and allow molecular study of preferential V-J-C association. It will provide a basis for a molecular study of preferential light chain expression (Lambda) in particular autoimmune diseases and the preferential use of the Lambda light chain with certain heavy chain classes. An additional specific objective is to assay for transcription enhancer elements associated with the above Ig genes. For these objectives we will clone rearranged and germline Ig Lambda genes by recombinant DNA techniques, insert these genes into plasmid vectors which have positive selection markers (Ecogpt or cat), transfect various lymphoid cells with these or deleted mutants, study expression of these genes by cellular immunofluorescence and electrophoresis of proteins and nucleic acids, and sequence particular DNA fragments by established procedures.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI021870-01
Application #
3132305
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1984-12-01
Project End
1987-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Miami School of Medicine
Department
Type
Schools of Medicine
DUNS #
City
Miami
State
FL
Country
United States
Zip Code
33101
Donohoe, M E; Blomberg, B B (1997) The 14.1 surrogate light chain promoter has lineage- and stage-restricted activity. J Immunol 158:1681-91
Glozak, M A; Blomberg, B B (1996) The human lambda immunoglobulin enhancer is controlled by both positive elements and developmentally regulated negative elements. Mol Immunol 33:427-38
Blomberg, B B; Glozak, M A; Donohoe, M E (1995) Regulation of human lambda light chain gene expression. Ann N Y Acad Sci 764:84-98
Yang, J; Glozak, M A; Blomberg, B B (1995) Identification and localization of a developmental stage-specific promoter activity from the murine lambda 5 gene. J Immunol 155:2498-514
Bauer Jr, T R; McDermid, H E; Budarf, M L et al. (1993) Physical location of the human immunoglobulin lambda-like genes, 14.1, 16.1, and 16.2. Immunogenetics 38:387-99
Bauer Jr, T R; Blomberg, B (1991) The human lambda L chain Ig locus. Recharacterization of JC lambda 6 and identification of a functional JC lambda 7. J Immunol 146:2813-20
Blomberg, B B; Rudin, C M; Storb, U (1991) Identification and localization of an enhancer for the human lambda L chain Ig gene complex. J Immunol 147:2354-8
Lopez, D M; Blomberg, B B; Padmanabhan, R R et al. (1989) Nuclear disintegration of target cells by killer B lymphocytes from tumor-bearing mice. FASEB J 3:37-43
Udey, J A; Blomberg, B B (1988) Intergenic exchange maintains identity between two human lambda light chain immunoglobulin gene intron sequences. Nucleic Acids Res 16:2959-69
Udey, J A; Blomberg, B (1987) Human lambda light chain locus: organization and DNA sequences of three genomic J regions. Immunogenetics 25:63-70

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