Steiner G, Koning F, Elbe A, Tschachler E, Yokoyama W M, Shevach E M, Stingl G, Coligan J E
Department of Dermatology I, University of Vienna Medical School, Austria.
Eur J Immunol. 1988 Sep;18(9):1323-8. doi: 10.1002/eji.1830180904.
Cell lines derived from Thy-1+ dendritic epidermal cells (Thy-1+DEC) display a marked heterogeneity in T cell receptor (TcR) expression including CD3-associated alpha/beta, C gamma 1/delta or C gamma 2/delta and C gamma 4/delta TcR. In order to investigate whether this heterogeneity is primarily imposed by in vitro culture conditions or, alternatively, is already present within the epidermis, we studied TcR expression by Thy-1+DEC in situ and on freshly isolated epidermal cell suspensions greatly enriched for Thy-1+DEC. Immunolabeling experiments showed that resident Thy-1+DEC are CD45+, Thy-1+, CD3+, TcR V beta 8-, CD5-, CD4- CD8-, CD25- lymphocytes. Immunoprecipitation of lysates from 125I-surface-labeled Thy-1+DEC-enriched epidermal cells with anti-CD3 epsilon, anti-C gamma 1,2,3 and anti-C gamma 4 antibodies, respectively, and subsequent analysis of the precipitates by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed only CD3-associated 35 kDa/45 kDa gamma/delta heterodimers. The demonstration of TcR heterodimers on resident Thy-1+DEC strongly implies that these cells are functional T cells. The selective expression of C gamma 1/delta (C57BL/6) and C gamma 1/delta or C gamma 2/delta (C3H/He) TcR makes these cells useful for the study of gamma/delta TcR function.
源自Thy-1⁺树突状表皮细胞(Thy-1⁺DEC)的细胞系在T细胞受体(TcR)表达上呈现出显著的异质性,包括与CD3相关的α/β、Cγ1/δ或Cγ2/δ以及Cγ4/δ TcR。为了研究这种异质性是主要由体外培养条件造成的,还是在表皮内就已存在,我们研究了原位以及新鲜分离的、富含Thy-1⁺DEC的表皮细胞悬液中Thy-1⁺DEC的TcR表达。免疫标记实验表明,驻留的Thy-1⁺DEC是CD45⁺、Thy-1⁺、CD3⁺、TcR Vβ8⁻、CD5⁻、CD4⁻、CD8⁻、CD25⁻淋巴细胞。分别用抗CD3ε、抗Cγ1、2、3和抗Cγ4抗体对¹²⁵I表面标记的富含Thy-1⁺DEC的表皮细胞裂解物进行免疫沉淀,随后通过二维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对沉淀物进行分析,结果仅显示与CD3相关的35 kDa/45 kDaγ/δ异二聚体。驻留的Thy-1⁺DEC上TcR异二聚体的证实强烈表明这些细胞是功能性T细胞。Cγ1/δ(C57BL/6)和Cγ1/δ或Cγ2/δ(C3H/He)TcR的选择性表达使这些细胞可用于γ/δ TcR功能的研究。