Abstract

Three projects concerning regulation of the expression of class I major histocompatibility genes in human lymphoblastoid cell lines (LCL) will be undertaken. (1) Which DNA sequences regulate transcription of the HLA-A, -B and -C genes? The class I genes in gamma ray-induced HLA antigen-loss mutants that have an apparently normal gene but lack the corresponding mRNA will be analyzed in order to answer this question. (2) The rate of transcription of class I genes in B-LCLS is approximately 20 times greater than that in T-LCLS. Which DNA sequences and proteins are responsible for this cell type-specific difference in transcription? Modified class I genes cloned in non-integrating (pHeBo) vectors will be tested for expression by transferring them into a class I-null B-LCL mutant and into class I-poor T-LCLs. Nuclease protection an migration retardation in gels will also be used to identify cell type-specific transcription regulatory sequences. Regulatory proteins will be sought on episomally maintained class I genes. (3) """"""""5.4"""""""", """"""""6.0"""""""" and """"""""6.2"""""""" are cloned, apparently normal, non-A, B, C class I genes for which corresponding antigens are unknown. They will be tested for expression, before and after in vitro modification,, by transfer into class I-null B-LCLs. If antigens appear antibodies will be raised against them and used to study their distribution in the body. Immunoprecipitation and electrophoresis will be used to study the biochemical abnormality in B-LCL mutants that have a post- transcriptional defect in class I antigen expression. Transfer of cloned class II genes into class II-null mutant B-LCLS will be used to create cells that have defined class II expression. These cells will then be used to study aspects of antigen presentation and T cell recognition. Some human B-LCLS are deficient in surface expression of the lymphocyte common antigen T200. The T200 DNA, RNA and protein of these mutants will be studied in order to determine the cause of T200 deficiency. Other T200 mutants will be isolated in ways to be informative about the genetic regulation of T200 expression.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI015486-14
Application #
3126196
Study Section
Immunobiology Study Section (IMB)
Project Start
1978-12-01
Project End
1992-11-30
Budget Start
1991-12-01
Budget End
1992-11-30
Support Year
14
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Kim, S K; DeMars, R (2001) Epitope clusters in the major outer membrane protein of Chlamydia trachomatis. Curr Opin Immunol 13:429-36
Ortiz, L; Angevine, M; Kim, S K et al. (2000) T-cell epitopes in variable segments of Chlamydia trachomatis major outer membrane protein elicit serovar-specific immune responses in infected humans. Infect Immun 68:1719-23
Kim, S K; Angevine, M; Demick, K et al. (1999) Induction of HLA class I-restricted CD8+ CTLs specific for the major outer membrane protein of Chlamydia trachomatis in human genital tract infections. J Immunol 162:6855-66
Ortiz, L; Demick, K P; Petersen, J W et al. (1996) Chlamydia trachomatis major outer membrane protein (MOMP) epitopes that activate HLA class II-restricted T cells from infected humans. J Immunol 157:4554-67
Ceman, S; Rudersdorf, R A; Petersen, J M et al. (1995) DMA and DMB are the only genes in the class II region of the human MHC needed for class II-associated antigen processing. J Immunol 154:2545-56
Ceman, S; Petersen, J W; Pinet, V et al. (1994) Gene required for normal MHC class II expression and function is localized to approximately 45 kb of DNA in the class II region of the MHC. J Immunol 152:2865-73
Celis, E; Tsai, V; Crimi, C et al. (1994) Induction of anti-tumor cytotoxic T lymphocytes in normal humans using primary cultures and synthetic peptide epitopes. Proc Natl Acad Sci U S A 91:2105-9
Malnati, M S; Ceman, S; Weston, M et al. (1993) Presentation of cytosolic antigen by HLA-DR requires a function encoded in the class II region of the MHC. J Immunol 151:6751-6
Sette, A; Ceman, S; Kubo, R T et al. (1992) Invariant chain peptides in most HLA-DR molecules of an antigen-processing mutant. Science 258:1801-4
Sanchez Perez, M; Diez Orejas, R; Petersen, J W et al. (1990) Differences in specificity of ""DPw2-specific"" cytotoxic T cell clones revealed with HLA-mutant lines: evidence that non-DP HLA genes influence recognition by some clones. Eur J Immunol 20:673-81

Showing the most recent 10 out of 28 publications