During terminal differentiation of B lymphocytes, natural killer (NK) cells recognize target structure (NKTS) expressed by (or uncovered on) B cells prior to the plasma cell stage. The objectives of this study are to identify and characterize NKTS expressed on lymphocytic target cells and to determine how expression of NKTS is regulated. NK-resistant lymphoid cell lines will be selected following low-dose irradiation of NK-sensitive lines, multiple rounds of NK, and cloning the surviving cells. The NK- resistant lines will be compared to the parental, sensitive lines by analytical, 2-D-polyacrylamide gel electrophoresis to detect polypeptide differences, i.e., candidate NKTS. Monoclonal and polyclonal antibody reagents will be produced for detailed analyses of the structure and function of candidate NKTS. One such molecule, p115, has been identified on the surface of an NK-sensitive, T-cell line and is absent from the NK- resistant, variant line. Antibody reagents of p115 will be produced for studies to determine its cellular distribution and function in NK cell- target cell binding. The expression of NKTS is regulated, in part, by MHC class I molecules. B-cell lines will be transfected with the E3 gene of Adenovirus; E3 encodes (among other polypeptides) a p19 that complexes with class I molecules, preventing their cell-surface expression. Transfected B-cell lines should show an increase in NK sensitivity, if NKTS are constitutively produced, but masked, by class I molecules. Alternatively, they should show no increase in sensitivity, if NKTS are synthesized de novo during B-cell termial differentiation. NK-sensitive, class I- deficient, B-cell lines will be transfected with different class I gene constructs to map the domain(s) interacting with NKTS. Polypeptides coprecipitating with MHC class I molecules will be purified and tested as candidate NKTS; a polypeptide has been identified that coprecipitates with class I molecules from a NK-sensitive line but not from the NK-resistant line. These studies should clarify whether NK cells regulate terminal differentiation of B cells (and interact with late-stage B-cell malignancies) and should identify both the phenotype of the NK-sensitive B cells and NKTS.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA042890-04
Application #
3184556
Study Section
Experimental Immunology Study Section (EI)
Project Start
1986-08-01
Project End
1994-05-31
Budget Start
1989-08-01
Budget End
1990-05-31
Support Year
4
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
Malyguine, A M; Scott, J E; Dawson, J R (1998) The role of calnexin in NK-target cell interaction. Immunol Lett 61:67-71
Hauser, T A; Malyguine, A M; Dawson, J R (1998) Conformation dependence of MHC class I in the modulation of target cell sensitivity to natural killing. Hum Immunol 59:71-6
Storkus, W J; Wei, M; Cresswell, P et al. (1996) Class I-like CD1A-C do not protect target cells from NK-mediated cytolysis. Cell Immunol 167:154-6
Malyguine, A M; Saadi, S; Platt, J L et al. (1996) Human natural killer cells induce morphologic changes in porcine endothelial cell monolayers. Transplantation 61:161-4
Scott, J E; Dawson, J R (1995) MHC class I expression and transport in a calnexin-deficient cell line. J Immunol 155:143-8
Scott, J E; Dawson, J R (1995) Heat treatment of leukemic cell lines can increase their sensitivity to NK lysis. Cell Immunol 163:296-302
Storkus, W J; Salter, R D; Cresswell, P et al. (1992) Peptide-induced modulation of target cell sensitivity to natural killing. J Immunol 149:1185-90
Wyatt, R M; Dawson, J R (1991) Characterization of a subset of human B lymphocytes interacting with natural killer cells. J Immunol 147:3381-8
Storkus, W J; Salter, R D; Alexander, J et al. (1991) Class I-induced resistance to natural killing: identification of nonpermissive residues in HLA-A2. Proc Natl Acad Sci U S A 88:5989-92
Storkus, W J; Alexander, J; Payne, J A et al. (1989) The alpha 1/alpha 2 domains of class I HLA molecules confer resistance to natural killing. J Immunol 143:3853-7

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