Wolos J A
Clin Immunol Immunopathol. 1986 Feb;38(2):222-34. doi: 10.1016/0090-1229(86)90140-6.
Supernatants from autologous mixed lymphocyte reaction (AMLR) cultures mediated the cytotoxic T-cell response in a system containing T cells and mitomycin C-treated, TNP-modified syngeneic thymocytes. No cytotoxic activity developed in the absence of the AMLR supernatants. The removal of thymic adherent cells abrogated the effect of the AMLR supernatants in the generation of cytotoxic cells. AMLR helper activity was restored following the addition of small numbers of splenic adherent cells. These results led to speculation that the AMLR supernatant interacted with accessory cells. Examination of the supernatant revealed significant levels of colony stimulating factor (CSF), and interferon-gamma (IFN-gamma). Functionally, incubation of the AMLR supernatant with P388D1, a macrophage tumor line, resulted in the production of interleukin 1 (IL-1). CSF is a major inducer of IL-1 synthesis. In addition, the AMLR supernatant induced the expression of Ia antigens on the P388D1 cells. IFN-gamma has been reported to mediate this activity. Although both lymphokines were present and appeared to have an effect on macrophages, it was unknown if both were required for cytotoxic T-cell development in this system. Samples of AMLR supernatant were incubated for 10 min at various temperatures. The ability of the AMLR supernatants to mediate cytotoxic T-cell activation was sensitive to incubation at 80 or 90 degrees C. Interestingly, at these temperatures CSF activity in the bone marrow colony-forming cell assay was enhanced. In separate experiments, AMLR culture supernatant was dialyzed against a pH 2 buffer. The induction of IA antigens by these supernatants was sensitive to dialysis at this pH. The similarity with IFN-gamma provides further evidence that this activity was mediated by IFN-gamma in the AMLR supernatants. The ability of the AMLR supernatants to mediate the cytotoxic T-cell response to altered self was also pH 2 sensitive. In contrast, IL-1 inducing capability and bone marrow colony growth, both CSF activities, were not reduced by pH 2 dialysis. Taken together, these data demonstrate a primary role for an IFN-like molecule, present in the AMLR supernatant, on cytotoxic T-cell activation. CSF involvement in this response is suggested by its IL-1 inducing activity, but cannot be definitively proven in the present study.
来自自体混合淋巴细胞反应（AMLR）培养物的上清液在一个包含T细胞以及经丝裂霉素C处理、经三硝基苯（TNP）修饰的同基因胸腺细胞的系统中介导了细胞毒性T细胞反应。在没有AMLR上清液的情况下未产生细胞毒性活性。去除胸腺黏附细胞消除了AMLR上清液在细胞毒性细胞生成中的作用。添加少量脾黏附细胞后，AMLR辅助活性得以恢复。这些结果引发了这样的推测，即AMLR上清液与辅助细胞相互作用。对上清液的检测显示出显著水平的集落刺激因子（CSF）和干扰素-γ（IFN-γ）。在功能上，将AMLR上清液与巨噬细胞肿瘤系P388D1一起孵育，导致白细胞介素1（IL-1）的产生。CSF是IL-1合成的主要诱导剂。此外，AMLR上清液诱导了P388D1细胞上Ia抗原的表达。据报道IFN-γ介导这种活性。尽管两种淋巴因子都存在且似乎对巨噬细胞有影响，但在该系统中细胞毒性T细胞的发育是否需要两者尚不清楚。将AMLR上清液样品在不同温度下孵育10分钟。AMLR上清液介导细胞毒性T细胞活化的能力对在80或90摄氏度下孵育敏感。有趣的是，在这些温度下，骨髓集落形成细胞测定中的CSF活性增强。在单独的实验中，将AMLR培养上清液用pH 2缓冲液透析。这些上清液对Ia抗原的诱导对在此pH下的透析敏感。与IFN-γ的相似性提供了进一步的证据，表明这种活性是由AMLR上清液中的IFN-γ介导的。AMLR上清液介导对改变的自身的细胞毒性T细胞反应的能力对pH 2也敏感。相比之下，IL-1诱导能力和骨髓集落生长，这两者都是CSF活性，并未因pH 2透析而降低。综上所述，这些数据证明了AMLR上清液中存在的一种类IFN分子在细胞毒性T细胞活化中起主要作用。CSF因其IL-1诱导活性而参与这种反应，但在本研究中无法得到确切证实。
