Kuchroo V K, Steele J K, Billings P R, Selvaraj P, Dorf M E
Department of Pathology, Harvard Medical School, Boston, MA 02115.
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9209-13. doi: 10.1073/pnas.85.23.9209.
Three suppressor T (Ts)-cell hybridomas specific for 4-hydroxy-3-nitrophenyl acetyl (NP) hapten were selected for surface expression of cluster determinant 3 (CD3) by using antibody (anti-CD3) or antigen (NP-bovine serum albumin) panning procedures followed by cloning at limiting dilution. The CD3-selected Ts hybridomas showed a 1-2 logarithmic enrichment in suppressor activity when compared to the parent lines; they also specifically bound NP-coupled sheep red blood cells in rosette assays. This antigen-binding ability could be down-modulated by anti-CD3 antibody. Similarly, surface expression of CD3 was specifically down-modulated by preincubation of these hybridomas with antigen. Anti-CD3 monoclonal antibody under reducing conditions coprecipitated a broad band of 38-50 kDa associated with two CD3 (25 and 16 kDa) bands. T-cell receptor, anti-alpha-specific monoclonal antibody also immunoprecipitated a broad band in the 41 to 49-kDa region. The combined results suggest that, like helper and cytotoxic T lymphocytes, Ts cells also bear antigen-specific receptors associated with CD3 molecules.
通过使用抗体(抗CD3)或抗原(NP-牛血清白蛋白)淘选程序,随后在有限稀释度下进行克隆,选择了三种对4-羟基-3-硝基苯基乙酰(NP)半抗原具有特异性的抑制性T(Ts)细胞杂交瘤,用于簇分化抗原3(CD3)的表面表达。与亲代细胞系相比,经CD3选择的Ts杂交瘤在抑制活性方面显示出1-2个对数级的富集;它们在花环试验中也能特异性结合NP偶联的绵羊红细胞。这种抗原结合能力可被抗CD3抗体下调。同样,通过用抗原预孵育这些杂交瘤,CD3的表面表达也会被特异性下调。在还原条件下,抗CD3单克隆抗体共沉淀出一条与两条CD3带(25 kDa和16 kDa)相关的38-50 kDa宽带。T细胞受体抗α特异性单克隆抗体也在41至49 kDa区域免疫沉淀出一条宽带。综合结果表明,与辅助性T淋巴细胞和细胞毒性T淋巴细胞一样,Ts细胞也带有与CD3分子相关的抗原特异性受体。