摘要
CIK是肿瘤过继性细胞免疫治疗中的免疫效应细胞。为使CIK在实验室里能被更有效地诱导增殖并赋予其更强的杀肿瘤效应,我们在CIK常规培养环境中加入抗CD28单抗和IL-15,探讨抗CD28单抗和IL-15对CIK增殖和杀肿瘤效应。取人外周血单个核细胞(PBMC),预先以常规方法诱导CIK,然后加入抗CD28单抗和IL-15与CIK共培养。用全自动五分类血液分析仪计数CIK增殖率;用流式细胞术测定CIK中粒酶B、穿孔素和CD107a等分子的表达率;用ELISA方法检测CIK分泌IL-10、IL-12、INF-γ和TNF-α水平;用乳酸脱氢酶释放法测定CIK对人肺癌细胞株(A549)、乳腺腺癌细胞株(MFC-7)和人黑素瘤细胞株(HME1)的杀伤活性。PBMC经常规CIK诱导培养以后再加入抗CD28单抗和IL-15与对照组比较,前者细胞增殖率明显增强(P<0.05);在CIK培养体系中加入抗CD28单抗和IL-15可促进颗粒酶B、穿孔素和CD107a等分子的表达率进一步增强(P<0.05);加入抗CD28单抗和IL-15,培养8d后CIK对A549、MFC-7和HME1细胞杀伤活性分别为82.2%、59.3%和70.6%,与对照组(分别为60.9%、49.6%和48.4%)相比差异有统计学意义(P<0.05);在培养体系中加入抗CD28单抗和IL-15,培养8d后其细胞因子IFN-γ、TNF-α分泌水平显著高于对照组(P<0.05),组间IL-10和IL-12的分泌量未见显著差异(P>0.05)。实验说明在CIK培养体系中加入抗CD28单抗和IL-15可增加CIK增殖率并提高其抗肿瘤效应。
CIKs are immune effector cells in cell biological cancer therapy. To improve effectively the capacity of inducing proliferation and anti-tumor activity of CIK in the lab, we added anti-CD28 antibody and IL-15 into the culture system, discussing anti-CD28 antibody and IL-15 on proliferation and anti-tumor effect of CIK. CIK was first cultured from peripheral blood mononuclear cell （PBMC） and then co-cultured with anti-CD28 antibody and IL-15. The proliferation rate of CIK was counted by fully automatic five categories of blood analyzer. The expressions of granzyme B, perforin, CD107a were detected by flow cytometry. Cytokines of IL-10, IL-12, INF-γand TNF-α were quantified by ELISA. Cytotoxicities of CIK on lung cancer cell line （A549）, breast adenocarcinoma cell line （MFC-7） and human melanoma cell line （HME1） were measured by lactate dehy- drogenase releasing assay. Adding anti-CD28 antibody and IL-15 into CIK conventional culture system could significantly increase proliferation rate and promote expression of granzyme B, perforin, and CD107a（P〈0.05）. The anti-tumor activity of CIK on 8 day after adding anti-CD28 antibody and IL-15 on A549, MFC-7 and HME1 were 82.2%, 59.3%, and 70.6M respectively, it was significantly different from control group（60.9 %, 49.6 % and 48.4 % ） （P〈0.05）. The secretion levels of IFN-γand TNF-α after co-culutre were also significantly higher than those in the control group（P〈0.05）, but there was no difference of IL-10 and IL-12 secretion levels between each group（P〉0.05）. This study demonstrated that adding anti-CD28 antibody and IL-15 into CIK conventional culture system could enhance CIK proliferation and anti-tumor activity.
出处
《现代免疫学》
CAS
CSCD
北大核心
2012年第6期484-489,共6页
Current Immunology
基金
南京军区医学科技创新课题(11MA040)
