Long term growth in vitro of human T cell blasts with maintenance of specificity and function.
Long term growth in vitro of human T cell blasts with maintenance of specificity and function.
复制标题
人类 T 细胞母细胞在体外长期生长并保持特异性和功能。
DOI：
10.4049/jimmunol.122.4.1255
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发表时间：
1979
影响因子：
4.4
通讯作者：
Hans Wigzell
中科院分区：
文献类型：
--
作者：
J. Kurnick;K. Grönvik;Arthur Kimura;J. Lindblom;Valdemar T. Skoog;O. Sjöberg;Hans Wigzell
通过抗原刺激和梯度离心纯化获得的人T淋巴细胞可以在体外保持永久分裂细胞，保留特异性和功能。通过重复添加从用 T 细胞有丝分裂原激活的体外淋巴细胞培养物中获得的上清液，可以避免持续存在抗原的需要，同时保持增殖。我们利用 PHA 刺激扁桃体淋巴细胞，并在 48 小时后收获上清液。此类上清液含有T母细胞的生长支持因子，并且这些因子与用于诱导生长因子的凝集素明显不同。因此，可以通过与琼脂糖珠偶联的抗凝集素免疫吸附剂从上清液中去除PHA，而不会对此类上清液的T细胞支持能力产生任何可检测到的负面影响。相反，虽然凝集素能够刺激新分离的淋巴细胞群，但去除凝集素后的上清液不会激活可检测数量的小淋巴细胞。在没有上清液因子的情况下，抗原特异性细胞也可以通过将 MLR 母细胞重新暴露于原始刺激细胞群，或通过在受辐射的同系细胞的陪伴下向母细胞中添加可溶性抗原 (PPD) 来维持。或者，同基因或异基因照射的细胞与有丝分裂原一起可以产生生长支持因子，从而维持胚细胞增殖。
Human T lymphoblasts obtained via antigenic stimulation and gradient centrifugation purification can be maintained as perpetual dividing cells in vitro, retaining specificity and function. By repeated addition of supernatants obtained from in vitro lymphocyte cultures activated with T cell mitogens, one can avoid the need of having antigen continually present, while maintaining proliferation. We have utilized PHA to stimulate tonsil lymphocytes and harvested supernatants after 48-hr. Such supernatants contain growth-supporting factors for T blasts, and these factors are clearly distinguishable from the lectin used to induce the growth factors. Thus, the PHA can be removed from the supernatants via an anti-lectin immunosorbent coupled to Sepharose beads, without having any detectable negative impact on the T blast supporting ability of such supernatants. Conversely, although the lectin is capable of stimulating freshly isolated populations of lymphocytes, the supernatant after lectin removal will not activate detectable numbers of small lymphocytes. In the absence of supernatant factors, antigen-specific cells can also be maintained either by reexposure of the MLR blasts to the original stimulator cell population, or by the addition of the soluble antigen (PPD) to blasts in the company of irradiated syngeneic cells. Alternatively, syngeneic or allogeneic irradiated cells together with mitogen can generate growth-supporting factors and thus sustain blast proliferation.