摘要
在体外使用CD40L作用于B淋巴细胞使之活化,通过测定B淋巴细胞表面分子及分泌细胞因子的变化,探讨B淋巴细胞作为抗原递呈细胞在抗肿瘤免疫中的优势,为B淋巴细胞特异性活化细胞毒T淋巴细胞(cytotoxic Tlymphocyte,CTL)进而发挥其肿瘤细胞杀伤效应奠定基础.取7名健康成人外周静脉血,经淋巴细胞分离液分离人外周血单个核细胞(PBMC),分为对照组和实验组,实验组培养液中加入加入CD40L和IL-4,共培养21 d.观察B淋巴细胞生长状况,并在第7 d、第14 d和第21 d,用流式细胞仪(FCAS)测定其表面分子CD80、CD86和CD19,在第21 d,使用酶联免疫吸附法(ELISA)测定培养液中的IL-12水平.结果显示,在CD40L的作用下,B淋巴细胞呈克隆样增殖,并高表达表面分子CD80/CD86和CD19,和对照组相比,B淋巴细胞分泌IL-12水平升高(P<0.05).上述实验结果说明通过CD40L的刺激,B淋巴细胞得到了活化,提示通过该途径活化的B淋巴细胞具备了作为抗原递呈细胞而发挥其抗肿瘤免疫治疗的能力.
Dendritic cells （DCs） constitute only 0. 1% - 0.5% of human peripheral blood mononuclear cells （PBMCs）, and in 2-3 weeks, DCs cease to proliferate and become less efficient in presenting antigen. Therefore, we chose CD40L to activate B lymphocytes in vitro and determined the expression of the surface molecular and IL-12 secretion of B lymphocytes to evaluate the advantage of B lymphocytes as Antigen Presenting Cells （APC） in antitumor immunity. Peripheral blood mononuclear cells （PBMCs） obtained from 7 healthy adult donors were isolated by Ficoll centrifugation. The experimental group PBMCs were cultured by RMPI 1640 supplemented with 10% fetal calf serum （FCS）, sCD40L （2 μg/mL） and recombinant human interleukin-4 （IL-4） （4 ng/mL） for 21 days. The proliferation of B lymphocytes was observed. In the day 7, 14 and 21, the expressions of CD80, CD86 and CD19 were determined respectively by flow cytometry analysis system（FCAS）. The level of interleukin-12（IL- 12） produced upon the cultured B lymphocytes was measured by using enzyme-linked immunosorbent assay （ELISA） on the day 21. The results show that the activated B lymphocytes were easily expandable and could from large clones and there was a high expression of CDSO/CD86 and CD19. The expression of CDS0 is 0. 05^（7 d）, 11. 94%（14 d） and 27.56%（21 d）, respectively. The expression of CD86 is 1.17%（7 d）, 9.51% （14 d） and 21.32% （21 d）, respectively. The expression of CD19 is 3. 03%（7 d）, 15. 23% （14 d） and 58. 66% （21 d）, respcetively. Comparing the results with that of the control group, we found that the level of IL-12 generated by B lymphocytes was up-regulated（P〈0.05）. These results show that B lymphoeytes may be used as alternative APC for the induction of antigen-specific CD8^＋ T cell（CTL） responses, which might be used in tumor immunotherapy.
出处
《南京大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第5期472-477,共6页
Journal of Nanjing University（Natural Science）
基金
国家自然科学基金(30471701)
