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[用纯化的刀豆球蛋白A刺激人外周血淋巴细胞的体外增殖反应及T细胞亚群分析(作者译)]

[The in vitro proliferative response of human peripheral blood lymphocytes stimulated with purified concanavalin A and the analysis of subpopulations of T-cells (author's transl)].

作者信息

Oh-Ishi T

出版信息

Hokkaido Igaku Zasshi. 1981 Jul;56(4):421-30.

PMID:6976924
Abstract

Human peripheral blood lymphocytes (PBL) were separated from anticoagulated peripheral blood using the Ficoll-Conraxin-L gradient technique, and were cultured with various concentrations of highly purified concanavalin A (Con A (F3)). Tritiated thymidine incorporation in these cultures was measured. PBL responded well to low doses of Con A (F3) and high doses of Con A (F3), and the dose-response curve to Con A (F3) of PBL seemed to show that bimodal proliferative response with a notch or a peak at 5--6 micrograms/ml of Con A (F3) and the other peak at the higher doses of Con A (F3). As it has been well known that E-rosette-forming cells (E-RFC) in human peripheral blood can be subdivided into two T-cell subpopulations on the basis of their relative affinity for sheep red blood cell (SRBC), we have examined how these T-cell subpopulations could responded to Con A (F3) at varying concentrations. We found that "Low-affinity E-RFC" could be optimally stimulated by Con A (F3) at 5--6 micrograms/ml, while "High-affinity E-RFC" at 8--12 micrograms/ml. 3H-thymidine incorporation of high-affinity E-RFC was strikingly increased at low doses of Con A (F3) when low-affinity E-RFC and high-affinity E-RFC were mixed at 25: 75 ratio, and the dose-response curve was very similar to that of PBL. Moreover when monocytes were added to high affinity E-RFC and the mixture was cultured, the doses-response curve was elevated at low doses of Con A (F3), but the dose-response curve was similar to that of high-affinity E-RFC and different from that of low-affinity E-RFC. Our results suggest that low-affinity E-RFC that has the capacity to respond optimally to Con A (F3) at 5--6 micrograms/ml in the in vitro proliferative response has the important effect upon the in vitro proliferative response of PBL to low doses of Con A (F3).

摘要

采用Ficoll-Conraxin-L梯度技术从抗凝外周血中分离出人外周血淋巴细胞(PBL),并将其与不同浓度的高纯度刀豆蛋白A(Con A (F3))一起培养。测量这些培养物中氚标记胸腺嘧啶核苷的掺入量。PBL对低剂量和高剂量的Con A (F3)反应良好,PBL对Con A (F3)的剂量反应曲线似乎显示出双峰增殖反应,在5 - 6微克/毫升的Con A (F3)处有一个凹口或峰值,在更高剂量的Con A (F3)处有另一个峰值。众所周知,人外周血中的E花环形成细胞(E-RFC)可根据其对绵羊红细胞(SRBC)的相对亲和力分为两个T细胞亚群,我们研究了这些T细胞亚群在不同浓度下对Con A (F3)的反应情况。我们发现,“低亲和力E-RFC”在5 - 6微克/毫升的Con A (F3)刺激下反应最佳,而“高亲和力E-RFC”在8 - 12微克/毫升的Con A (F3)刺激下反应最佳。当低亲和力E-RFC和高亲和力E-RFC以25:75的比例混合时,低剂量的Con A (F3)会显著增加高亲和力E-RFC的3H-胸腺嘧啶核苷掺入量,且剂量反应曲线与PBL的非常相似。此外,当将单核细胞添加到高亲和力E-RFC中并进行培养时,低剂量的Con A (F3)会使剂量反应曲线升高,但该剂量反应曲线与高亲和力E-RFC的相似,与低亲和力E-RFC的不同。我们的结果表明,在体外增殖反应中对5 - 6微克/毫升的Con A (F3)有最佳反应能力的低亲和力E-RFC,对PBL对低剂量Con A (F3)的体外增殖反应具有重要影响。

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