Doroszczak N, Yoshida T, Cohen S
J Immunol. 1977 Nov;119(5):1617-20.
Identification of a subpopulation of lymphocytes producing lymphokines was attempted by fractionating the lymph node cells from guinea pigs immune to DNP-BSA by velocity sedimentation at 1 x G. Each of six fractions obtained by this procedure was cultured with or without the presence of antigen, and the culture supernatants that were separated 24 hr later were assayed for various lymphokine activities. Most of the lymphokines, including migration inhibition factor, chemotactic factor for neutrophils, mitogenic factor, and lymphotoxin were generated by the first two fractions of lymphocytes, which represented the largest, most rapidly sedimenting cells. Although th procedure of cell separation does not depend on cell surface properties, the larger cells contained more cells with T cell surface markers and the smalller contained more cells with B cell surface markers. Proliferative response of those lymphocytes measured by 3H-thymidine uptake, however, has shown that the largest two subpopulations responded poorly either to specific antigens or to mitogens (PHA and LPS), and rather that the medium size cells responded most strongly to the both stimulants. These results indicated that the production of some lymphokines confined to certain subpopulations of lymphocytes which are large in size. Further, these cells are readily separable from the medium sized cells that respond strongly to antigenic and mitogenic stimuli with mitogenic responses.
通过在1×G下进行速度沉降，对免疫了二硝基苯牛血清白蛋白（DNP-BSA）的豚鼠的淋巴结细胞进行分级分离，试图鉴定产生淋巴因子的淋巴细胞亚群。通过该程序获得的六个级分中的每一个，都在有或没有抗原存在的情况下进行培养，24小时后分离得到的培养上清液用于检测各种淋巴因子活性。大多数淋巴因子，包括迁移抑制因子、嗜中性粒细胞趋化因子、促有丝分裂因子和淋巴毒素，都是由淋巴细胞的前两个级分产生的，这两个级分代表了最大、沉降最快的细胞。尽管细胞分离程序不依赖于细胞表面特性，但较大的细胞含有更多具有T细胞表面标志物的细胞，而较小的细胞含有更多具有B细胞表面标志物的细胞。然而，通过3H-胸腺嘧啶核苷摄取测量的这些淋巴细胞的增殖反应表明，最大的两个亚群对特异性抗原或有丝分裂原（PHA和LPS）的反应都很差，相反，中等大小的细胞对这两种刺激的反应最强。这些结果表明，某些淋巴因子的产生局限于某些较大尺寸的淋巴细胞亚群。此外，这些细胞很容易与对抗原和有丝分裂原刺激有强烈有丝分裂反应的中等大小细胞分离。
