The objectives of this proposal are to study the CH series B cell lymphomas which arose in B10.H-2aH-4bp/Wts mice and the normal Ly-1+ B cells from which they are derived. The CH lymphomas will be employed as models to study B cell differentiation. We will use tissue-culture adapted subclones of lymphomas, whose cells bear membrane IgM reactive with known antigens. We will concentrate on a group of tumors reactive with membrane phospholipids such as phosphatidyl choline, recognizable by the tumor IgM on sheep erythrocytes and bromelain treated mouse erythrocytes. We will study the influence of specific antigen, mitogens, lymphokines and MHC restricted T helper cells on the differentiation of the B cells. Differentiation will be detected as secretion of IgM, by localized hemolysis, or as a change in the immunoglobulin isotype produced. The latter will be detected by flow cytometry and by isotype specific ELISA of the supernatant from cultures grown to high density. We will sort and clone variant cells producing the changed isotypes and will compare the specificity and idiotype of the Ig produced with that of the IgM produced by the parent tumor. We will determine whether the same VH and VL genes are used for the parental IgM and the switched isotype. We will use restriction digest analysis of DNA, sandwich hybridization of RNA and nucleotide sequencing to elucidate the genetic mechanisms responsible for iusotype switching. We will use tissue culture, flow cytometry and ELISA to investigate the nature of signals which control isotype switching. We will also compare the specificity and idiotype of immunoglobulins produced by hybridomas derived from normal Ly- 1+B cells with those expressed by 27 previously characterized CH lymphomas. The objectives is to determine whether the CH lymphomas are derived from a random selection of the normal Ly- 1+B cells of syngeneic mice and to determine whether the Ig produced by normal Ly-1+B cells is as limited in specificity and idiotype as is that of the CH lymphomas. We will undertake exploratory experiments to test the importance of idiotype/antiidotype regulation in controlling the normal population of Ly-1+B cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA023770-10
Application #
3166236
Study Section
Immunobiology Study Section (IMB)
Project Start
1979-01-01
Project End
1991-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
10
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Whitmore, A C; Prowse, D M; Haughton, G et al. (1991) Ig isotype switching in B lymphocytes. The effect of T cell-derived interleukins, cytokines, cholera toxin, and antigen on isotype switch frequency of a cloned B cell lymphoma. Int Immunol 3:95-103
Pennell, C A; Maynard, E; Arnold, L W et al. (1990) High frequency expression of S107 VH genes by peritoneal B cells of B10.H-2aH-4bP/WTS mice. J Immunol 145:1592-7
Bishop, G A; Pennell, C A; Travis, W et al. (1990) Antibodies specific for Ig idiotype, but not isotype, can substitute for antigen to induce IgM secretion by a B cell clone. Int Immunol 2:285-90
Van Houten, N; Haughton, G (1990) Simultaneously arising Ly-1(CD5) B cell lymphomas have identical expressed IgH and kappa-genes but different nonproductive heavy chain rearrangements. J Immunol 144:745-51
Whitmore, A C; Prowse, D M; Arnold, L W et al. (1989) Ig isotype switching in B lymphocytes. A method for estimating isotype switch frequency in cloned B cell lymphomas. Int Immunol 1:532-9
Van Houten, N; Willoughby, P B; Arnold, L W et al. (1989) Early commitment to neoplasia in murine B- and T-cell lymphomas arising late in life. J Natl Cancer Inst 81:47-54
Pennell, C A; Sheehan, K M; Brodeur, P H et al. (1989) Organization and expression of VH gene families preferentially expressed by Ly-1+ (CD5) B cells. Eur J Immunol 19:2115-21
Mercolino, T J; Locke, A L; Afshari, A et al. (1989) Restricted immunoglobulin variable region gene usage by normal Ly-1 (CD5+) B cells that recognize phosphatidyl choline. J Exp Med 169:1869-77
Pennell, C A; Mercolino, T J; Grdina, T A et al. (1989) Biased immunoglobulin variable region gene expression by Ly-1 B cells due to clonal selection. Eur J Immunol 19:1289-95
Arnold, L W; Grdina, T A; Whitmore, A C et al. (1988) Ig isotype switching in B lymphocytes. Isolation and characterization of clonal variants of the murine Ly-1+ B cell lymphoma, CH12, expressing isotypes other than IgM. J Immunol 140:4355-63

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