CD3、CD28抗体增强淋巴细胞转化功能的研究

Effect of anti-CD3/28 on enhancing lymphocyte transform function

目的:研究CD3、CD28抗体对淋巴细胞转化的增强作用,以期了解与评价应用外周血单个核细胞(PBMC)体外扩增淋巴细胞时双抗体作用特性及抗原提呈细胞(APC)数量对扩增的影响。方法:利用淋巴细胞转化实验3H-胸腺嘧啶(3H-TdR)掺入法、细胞培养技术及流式细胞术(FCM),以包被CD3抗体、CD28抗体诱导PBMC中淋巴细胞转化,增加抗原提呈细胞(APC)浓度,以CD3抗体刺激PBMC(正常APC)中淋巴细胞转化为对照,以各组刺激指数(SI)来计算各因素对淋巴细胞转化的影响,及分析CD8+CD25+细胞占总转化细胞的百分比变化。结果:正常APC浓度的淋巴细胞转化组,CD3抗体可增加淋巴细胞转化,CD28抗体有协同作用;去APC组,CD3抗体增强淋巴细胞转化作用明显下降,CD28抗体仍有协同作用;4倍APC组,CD3抗体可以明显增强淋巴细胞转化作用,CD28抗体协同作用不明显。CD28抗体单独无增加淋巴细胞转化作用。流式细胞仪结果与SI结果平行。结论:CD28抗体可协同CD3抗体模拟体内第一、二信号增强诱导淋巴细胞转化,CD28可替代APC的第二信号作用,但APC有其它独立活化淋巴细胞的作用。

Objective： To study the effect of anti-CD3 CD28 on enhancing lymphocyte transform function （LTF）, in order to understand and evaluate characteristics of anti-CD3/28 to expand T lymphocytes in PBMC and affection of the APC amount to T cells expanding. Methods： with the methods of incorporation of ^3H-thymidine （^3H-TdR） for measure LTF, cell culture and flow cytometry （FCM）, using coated anti-CD3 and anti-CD28 to stimulate T lymphocytes. Then raised the antigen presentation cell （APC） concentration, measured the LTF with normal APC concentration group of anti-CD3 alone as control. SI and CD8^＋ CD25^＋ cells＇ percentage was used to evaluate all factors to LTF. Results： anti-CD3 can enhance LTF in normal APC grounp and anti-CD28 has co-stimulation function. In no APC group, the LTF of anti-CD3 came down sharply, but anti-CD28 also had costimulation function. In four times APC group, anti-CD3 can enhance LTF in normal APC grounp more and anti-CD28 had no costimulation function. Only anti-CD28 had no effective activity. The result of FCM was parallel with the SI. Conclusion： anti-CD3 and anti-CD28 can mimic first and second signal to active T cells, while anti-CD28 may replace second signal function which offered by APC. But APC also has other function to affect lymphocytes activity.