Abstract

This research is directed towards obtaining detailed information on the structural properties of the antigen-specific surface component of defined murine and primate T-cell lines. The overall goal is to obtain detailed information regarding the biochemical and molecular biological properties of T-cell surface components which are serologically related to immunoglobulin-V?H? region markers (V?T?), which will allow direct comparison with other possible T-cell receptors such as surface complexes restricted by the major histocompatibility complex.
The specific aims are: (1) to characterize the V?T?-related T-cell molecules structurally. The major approach here will be to begin with large quantities of homogenous, in vitro-grown T-cell lines (greater than 50 grams) and to prepare the molecules by immune affinity chromatography. Conditions for specific cleavage of peptide bonds will be used and the fragments will be separated by high performance liquid chromatography. (2) The restricted V?T?-related determinant expressed and synthesized by the T-cell line 70-N2 is inherited as an autosomal dominant within human populations. We will determine the genetic linkage of the gene specifying this marker. (3) In order to ensure that complete sequence information is obtained, we will approach the problem of the V?T?-bearing t-cell receptor of our defined lines using the techniques of molecular biology. We will use our specific antisera to isolate intact polyribosomes bearing nascent polypeptide chains from our cell lines, isolate mRNA and produce cDNA followed by cloning and sequencing of the cDNA. An alternative strategy will be the construction of a cDNA library using total mRNA and the expression vector pUC8 which allows synthesis of antigenic products. We will screen the cDNA library with nicktranslated DNA probes directed against putative T-cell surface receptors. The positive clones will then be screened with our antisera to determine whether the antisera we have generated to V?T?-related products react with the products of the putative T-cell receptor genes recently sequenced. (CS)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA042049-05
Application #
3182827
Study Section
(SSS)
Project Start
1985-09-01
Project End
1988-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
5
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Medical University of South Carolina
Department
Type
Schools of Medicine
DUNS #
183710748
City
Charleston
State
SC
Country
United States
Zip Code
29425

  Publications

Bhattacharya, P (1998) beta-specific mRNA in suppressor T-cell line and hybridoma that do not synthesize beta-polypeptide. Indian J Exp Biol 36:520-2
Dehghanpisheh, K; Huang, D; Schluter, S F et al. (1995) Production of IgG autoantibodies to TCRs in mice infected with the retrovirus LP-BM5. Int Immunol 7:31-6
Watson, R R; Wang, J Y; Dehghanpisheh, K et al. (1995) T cell receptor V beta complementarity-determining region 1 peptide administration moderates immune dysfunction and cytokine dysregulation induced by murine retrovirus infection. J Immunol 155:2282-91
Kurt, R A; Park, J A; Panelli, M C et al. (1995) T lymphocytes infiltrating sites of tumor rejection and progression display identical V beta usage but different cytotoxic activities. J Immunol 154:3969-74
Schluter, S F; Wang, E; Winfield, J B et al. (1995) Autoregulation of Tcr V region epitopes in autoimmune disease. Adv Exp Med Biol 383:231-6
Marchalonis, J J; Hohman, V S; Kaymaz, H et al. (1994) Cell surface recognition and the immunoglobulin superfamily. Ann N Y Acad Sci 712:20-33
Kaymaz, H; Dedeoglu, F; Schluter, S F et al. (1994) Cross-reactions of anti-immunoglobulin sera with synthetic T-cell receptor beta peptides: mapping on a 3-dimension model. Adv Exp Med Biol 347:93-8
Lake, D F; Schluter, S F; Wang, E et al. (1994) Autoantibodies to the alpha/beta T-cell receptors in human immunodeficiency virus infection: dysregulation and mimicry. Proc Natl Acad Sci U S A 91:10849-53
Marchalonis, J J; Schluter, S F; Wang, E et al. (1994) Synthetic autoantigens of immunoglobulins and T-cell receptors: their recognition in aging, infection, and autoimmunity. Proc Soc Exp Biol Med 207:129-47
Lake, D F; Bernstein, R M; Hersh, E M et al. (1994) Construction and serological characterization of a recombinant human single chain T cell receptor. Biochem Biophys Res Commun 201:1502-9

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