Abstract
We have used antibody-mediated complement killing to isolate membrane IgM-negative (mIgM−) variants from the mIgM+ murine B cell lymphoma, WEHI 279.1. This procedure has been used previously to select variants which lack expression of other cell-surface antigens on lymphoid cells. In those experiments, multiple rounds of selection have often been required for selection of the negative variants. We found that many cycles of selection produced very few variants and that those isolated had reduced, but still measurable, levels of mIgM. We were able to select large numbers of stable mIgM− variants by subjecting the populations with reduced levels of mIgM to two rounds of immunoselection within one cell cycle. These variants are stable and exhibit a variety of defects which are all expressed as a failure to display IgM on their external surface. Analysis of these variant clones at the biochemical level will begin to define the requirements for proper display of mIgM on the cell membrane of B lymphoma cells.
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Literature cited
Scharff, M.D. (1975).Harvey Lect. 69:125–142.
Coffino, P., and Scharff, M.D. (1971).Proc Natl. Acad. Sci. U.S.A. 68:219–225.
Secher, D., Milstein, C., and Adetugbo, K. (1977).Immunol. Rev. 23:31–72.
Holtkamp, B., Cramer, M., Lenake, H., and Rajewsky, K. (1981).Nature 289:66–68.
Francus, T., and Birshtein, B. (1978).Biochemistry 17:4324–4331.
Choi, E., Kuehl, and Wall, R. (1980).Nature 286:776–779.
Seidman, J.G., and Leder, P. (1980).Nature 286:779–783.
Warner, N. (1974).Adv. Immunol. 19:67–216.
Pious, D., Hawkey, P., and Forrest, G. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:1397–1400.
Hyman R. (1973).J. Natl. Cancer Inst. 50:415–422.
Rajan, T.V. (1977).Immunogenetics 4:105–113.
Pious, D., Soderland, C., and Gladstone, P. (1977).Immunogenetics 4:437–448.
Andreotti, P., Apgar, J., and Cresswell, P. (1980).Hum. Immunol. 1:77–86.
Bourguignon, L., Hyman, R., Trowbridge, I.S., and Singer, S.J. (1978).J. Exp. Med. 147:1348–1357.
Gladstone, P., Fuerez, L., and Pious, D. (1982).Proc. Natl. Acad. Sci. U.S.A. 79:1235–1239.
Lanier, L.L., and Warner, M.L. (1981).J. Immunol. 126:626–631.
Kruth, H.S., Braylan, R.C., Benson, N.A., and Nourse, V.A. (1981).Cancer Res. 41:4895–4899.
Brown, J., Klitzman, J., and Hellstrom, K.E. (1977).J. Immunol. Methods 15:57–66.
Glassy, M. (1978). PhD thesis, University of California, Riverside.
Sibley, C.H., Ewald, S., Kehry, M., Douglas, R., Raschke, W., and Hood, L. (1980).J. Immunol. 125:2097–2103.
Dragsten, P., Hankart, P., Blumenthal, J., Weinstein, J., and Schlessinger, J. (1979).Proc. Natl. Acad. Sci. U.S.A. 76:5163–5168.
Schreiner, G.F., Fujiwara, K., Pollard, T.D., and Unanue, E.R. (1977).J. Exp. Med. 145: 1393–1402.
Braun, J., Fujiwara, K., Pollard, T.D., and Unanue, E.R. (1978).J. Cell Biol. 79:409–418.
Braun, J., Hochman, P.S., and Unanue, E.R. (1982).J. Immunol. 128:1198–1204.
Podack, E.R., Biesecker, G., and Müller-Eberhard, H.J. (1979).Proc. Natl. Acad. Sci. U.S.A. 76:897–906.
Sibley, C., Andrews-Wagner, R. (1983) Immunogenetics (in press).
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This work was supported by NIH grant CA29679, ACS grant IM302, and American Cancer Society Faculty Research Award (FRA-178), all awarded to C.H.S.
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Andrews-Wagner, R., Sibley, C.H. Selection of membrane IgM− variants from a mIgM+ murine B lymphoma cell: Problems and solutions. Somat Cell Mol Genet 9, 43–54 (1983). https://doi.org/10.1007/BF01544047
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DOI: https://doi.org/10.1007/BF01544047