Morecki S, Topalian S L, Myers W W, Okrongly D, Okarma T B, Rosenberg S A
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
J Biol Response Mod. 1990 Oct;9(5):463-74.
A direct positive panning technique was developed in order to achieve highly purified CD4+ and CD8+ cells. Fresh peripheral blood mononuclear cells or tumor-infiltrating lymphocytes derived from bulk cultures were applied to modified polystyrene surfaces to which anti-CD4 or anti-CD8 monoclonal antibodies were covalently attached. Adherent cells were allowed to grow in the original flask and were then harvested and cultured in IL-2-containing medium. This positive immunoselection technique resulted in CD4+ and CD8+ cell subsets with high cell viability and a high degree of purity. In several samples, the isolated cell subsets were subsequently subjected to a negative immunomagnetic bead selection in order to remove reciprocal cell subset contamination or double-positive CD4+/CD8+ cells. The isolated cells maintained their ability to proliferate, kept their phenotypic profiles, and remained functionally intact after long-term growth in culture without the further addition of mitogenic or allogeneic cell stimulation. This approach is a simple, rapid, and reproducible technique that might be useful on a large scale to isolate and to grow T-cell subsets for research and for clinical use.
为了获得高度纯化的CD4+和CD8+细胞，开发了一种直接阳性淘选技术。将来自大量培养物的新鲜外周血单核细胞或肿瘤浸润淋巴细胞应用于共价连接有抗CD4或抗CD8单克隆抗体的改良聚苯乙烯表面。使贴壁细胞在原培养瓶中生长，然后收获并在含白细胞介素-2的培养基中培养。这种阳性免疫选择技术产生了具有高细胞活力和高纯度的CD4+和CD8+细胞亚群。在几个样本中，随后对分离的细胞亚群进行阴性免疫磁珠选择，以去除相互的细胞亚群污染或双阳性CD4+/CD8+细胞。分离的细胞保持其增殖能力，保持其表型特征，并且在无进一步添加促有丝分裂或同种异体细胞刺激的情况下在培养中长期生长后仍保持功能完整。这种方法是一种简单、快速且可重复的技术，可能在大规模分离和培养T细胞亚群用于研究和临床应用方面有用。
