Itoh K, Balch C M, Platsoucas C D
Department of General Surgery, University of Texas System Cancer Center, Houston 77030.
Cell Immunol. 1988 Aug;115(1):36-56. doi: 10.1016/0008-8749(88)90160-8.
We report a new, monocyte-independent system for the induction of activation and proliferation of human T cells in response to murine hybridomas expressing the OKT3 monoclonal antibody (OKT3 hybridomas). Incubation of nylon-wool-nonadherent (NA) lymphocytes or purified T cells with OKT3 hybridomas resulted in interleukin-2 (IL-2) production, expression of IL-2 receptor, modulation of the CD3 antigen, and proliferation. In contrast, murine hybridomas (OKT4, OKT8, anti-HLA-DR, and others) expressing monoclonal antibodies (mAb) other than OKT3 did not induce T-cell activation and proliferation. T cells did not respond to OKT3 mAb alone. OKT3 hybridomas alone did not produce interleukin-1 (IL-1) or other soluble factors that might be involved in the induction of IL-2 production by T cells, and they did not contain membrane-bound IL-1. In addition, IL-1 activity was not detected in cultures of NA-lymphocytes and OKT3 hybridomas, clearly demonstrating that IL-1 was not required, at least in this system, for T-cell activation and proliferation. Direct cell-cell contact between T cells and OKT3 hybridomas was required for IL-2 production. Thirty to fifty percent of T cells formed conjugates with the OKT3 hybridomas but not with the OKT4 or OKT8 hybridomas. Both conjugate formation and IL-2 production were significantly inhibited by the OKT3 mAb and by the anti-LFA-1 mAb. The cells responsible for IL-2 production were found to be of the T3+ T4+ T8- Leu 7- Leu 11- phenotype. IL-2 activity produced by NA-lymphocytes in response to OKT3 hybridomas became detectable as early as 1 hr and reached a maximum by 8 hr, preceding IL-2 receptor expression, modulation of the CD3 antigen, and [3H]thymidine incorporation of T cells. T cells produced higher concentrations of IL-2 in response to OKT3 hybridomas than in response to equal numbers of monocytes and OKT3 mAb. Addition of monocytes to cultures of T cells and OKT3 hybridomas resulted in suppression of IL-2 production in a concentration-dependent manner, suggesting that monocytes regulate the levels of IL-2 production. This monocyte-independent system may be useful for further dissection of T-cell activation and proliferation and its regulation by monocytes.
我们报告了一种新的、不依赖单核细胞的系统，用于诱导人类T细胞在响应表达OKT3单克隆抗体的鼠杂交瘤（OKT3杂交瘤）时的激活和增殖。将尼龙毛非黏附（NA）淋巴细胞或纯化的T细胞与OKT3杂交瘤一起孵育，会导致白细胞介素-2（IL-2）的产生、IL-2受体的表达、CD3抗原的调节以及细胞增殖。相比之下，表达除OKT3之外的单克隆抗体（mAb）的鼠杂交瘤（OKT4、OKT8、抗HLA-DR等）不会诱导T细胞激活和增殖。T细胞对单独的OKT3 mAb无反应。单独的OKT3杂交瘤不会产生白细胞介素-1（IL-1）或其他可能参与T细胞诱导产生IL-2的可溶性因子，并且它们不含有膜结合的IL-1。此外，在NA淋巴细胞和OKT3杂交瘤的培养物中未检测到IL-1活性，这清楚地表明，至少在该系统中，T细胞激活和增殖不需要IL-1。IL-2的产生需要T细胞与OKT3杂交瘤之间直接的细胞-细胞接触。30%至50%的T细胞与OKT3杂交瘤形成结合物，但不与OKT4或OKT8杂交瘤形成结合物。结合物的形成和IL-2的产生均受到OKT3 mAb和抗LFA-1 mAb的显著抑制。发现负责产生IL-2的细胞具有T3+ T4+ T8- Leu 7- Leu 11-表型。NA淋巴细胞响应OKT3杂交瘤产生的IL-2活性早在1小时就可检测到，并在8小时达到最大值，早于IL-2受体表达、CD3抗原的调节以及T细胞的[3H]胸苷掺入。与等量的单核细胞和OKT3 mAb相比，T细胞对OKT3杂交瘤产生的IL-2浓度更高。将单核细胞添加到T细胞和OKT3杂交瘤的培养物中会导致IL-2产生以浓度依赖的方式受到抑制，这表明单核细胞调节IL-2的产生水平。这种不依赖单核细胞的系统可能有助于进一步剖析T细胞激活和增殖及其受单核细胞的调节。
