Characterization of lymphocyte function-associated antigen 1 (LFA-1)-deficient T cell lines: the alphaL and beta2 subunits are interdependent for cell surface …

KS Weber, MR York, TA Springer… - Journal of immunology …, 1997 - journals.aai.org
KS Weber, MR York, TA Springer, LB Klickstein
Journal of immunology (Baltimore, Md.: 1950), 1997journals.aai.org
The leukocyte, or beta2, integrins are heterodimeric cell surface molecules that share a
common beta subunit, and have unique alpha subunits. LFA-1 is the predominant beta2
integrin on lymphocytes, and plays an important role in many lymphocyte functions;
however, most studies of the cytoplasmic regions of LFA-1 have been performed in
transfected epithelial cells, such as COS, in part because no lymphoid cell lines deficient in
the LFA-1 alpha subunit have been described. To address structure-function studies of …
Abstract
The leukocyte, or beta2, integrins are heterodimeric cell surface molecules that share a common beta subunit, and have unique alpha subunits. LFA-1 is the predominant beta2 integrin on lymphocytes, and plays an important role in many lymphocyte functions; however, most studies of the cytoplasmic regions of LFA-1 have been performed in transfected epithelial cells, such as COS, in part because no lymphoid cell lines deficient in the LFA-1 alpha subunit have been described. To address structure-function studies of beta2 integrins in relevant cell types, two T lymphoblastoid cell clones that completely lack cell surface LFA-1, J-(beta2).7 and SK-(beta2).7, derived from Jurkat and SKW3, respectively, were prepared by chemical mutagenesis and selection. Biosynthetic labeling and immunoprecipitation showed that the J-(beta2).7 clone did not translate any LFA-1 alpha subunit protein, while the SK-(beta2).7 cells did not synthesize any beta2 subunit protein. Northern blot analysis of poly(A+) RNA from these cells revealed an absence of the corresponding mRNA in each case. By transfection analysis, only the alpha subunit reconstituted LFA-1 expression in the J-(beta2).7 cells, while only the beta subunit restored cell surface LFA-1 expression in the SK-(beta2).7 cells. Functional studies with the parental cell lines, the J-(beta2).7 and SK-(beta2).7 cells, and the transfectants showed that all binding of Jurkat and SKW3 cells to purified ICAM-1 is mediated by LFA-1, and the reconstituted LFA-1 expressed by the J-(beta2).7 and SK-(beta2).7 transfected cells is regulated normally.
journals.aai.org