Alvarez J, Rodríguez J, López-Botet M, Silva A, De Landazuri M O
J Immunol Methods. 1981;40(3):289-96. doi: 10.1016/0022-1759(81)90360-4.
Supernatants of human PBL cultures stimulated with PHA contain a lymphokine named T-cell growth factor (TCGF). To remove the contaminant PHA from these supernatants, we have designed a simple and effective method based on the binding properties of this lectin to chicken red blood cells. This procedure completely removes the functional activities of a PHA solution, both in lectin-induced cellular cytotoxicity against erythroid targets and in DNA synthesis of fresh PBL. The TCGF activity remaining in the supernatants after absorption with CRBC is dependent on the cellular concentration employed in the preparation of the supernatants. The absorption procedure does not remove significant TCGF activity in the supernatants prepared at high cellular concentrations (5 X 10(6) cells/ml), whereas in those prepared at low cellular concentrations (1 X 10(6) cells/ml) a partial loss of TCGF activity is detected.
用PHA刺激的人外周血淋巴细胞(PBL)培养上清液中含有一种名为T细胞生长因子(TCGF)的淋巴因子。为了从这些上清液中去除污染物PHA,我们基于这种凝集素与鸡红细胞的结合特性设计了一种简单有效的方法。该程序完全消除了PHA溶液的功能活性,无论是在凝集素诱导的针对红细胞靶标的细胞毒性方面,还是在新鲜PBL的DNA合成方面。用鸡红细胞吸收后,上清液中残留的TCGF活性取决于制备上清液时所用的细胞浓度。在高细胞浓度(5×10⁶个细胞/毫升)制备的上清液中,吸收程序不会去除显著的TCGF活性,而在低细胞浓度(1×10⁶个细胞/毫升)制备的上清液中,会检测到TCGF活性部分丧失。