Kreuzfelder E, Menne S, Ferencik S, Roggendorf M, Grosse-Wilde H
Institut für Immunologie, Universitätsklinikum-GHS Essen, Essen, Germany.
Clin Immunol Immunopathol. 1996 Mar;78(3):223-7. doi: 10.1006/clin.1996.0033.
The levels of incorporated exogenous [3H]thymidine of peripheral blood mononuclear cells (PBMC) of the woodchuck were low after stimulation with mitogens concanavalin A (ConA), phytohemagglutinin (PHA), and pokeweed mitogen (PWM) when compared with other cell systems. The use of EDTA as an anticoagulant for blood sampling and AIM-V medium for culturing of PBMC improved the [3H]thymidine uptake of PBMC. A pronounced uptake is observed after use of [3H]adenine instead of [3H]thymidine for PBMC proliferation measurement. One likely explanation for the difference in [3H]adenine versus [3H]thymidine uptake is that the alternative pathway for thymidine monophosphate synthesis is important: the conversion of uridine to uridine monophosphate and, thereafter, to thymidine monophosphate. The optimal conditions for mitogen-induced proliferation of PBMC of the woodchuck were 2 micrograms/ml ConA and PHA at day 4 and 0.14 micrograms of PWM/ml at day 5. No consistent differences of [3H]adenine uptake were observed between PBMC from four woodchuck hepatitis virus-infected woodchucks and five uninfected animals.
与其他细胞系统相比，土拨鼠外周血单个核细胞（PBMC）在用丝裂原刀豆球蛋白A（ConA）、植物血凝素（PHA）和商陆有丝分裂原（PWM）刺激后，掺入的外源性[3H]胸腺嘧啶核苷水平较低。使用乙二胺四乙酸（EDTA）作为血液采样的抗凝剂以及AIM-V培养基培养PBMC可提高PBMC对[3H]胸腺嘧啶核苷的摄取。在用[3H]腺嘌呤代替[3H]胸腺嘧啶核苷进行PBMC增殖测量后，观察到明显的摄取。[3H]腺嘌呤与[3H]胸腺嘧啶核苷摄取差异的一个可能解释是，胸苷单磷酸合成的替代途径很重要：尿苷转化为尿苷单磷酸，然后再转化为胸苷单磷酸。土拨鼠PBMC丝裂原诱导增殖的最佳条件是第4天2微克/毫升ConA和PHA，第5天0.14微克/毫升PWM。在4只感染土拨鼠肝炎病毒的土拨鼠和5只未感染动物的PBMC之间，未观察到[3H]腺嘌呤摄取的一致差异。
