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目的探讨高强度聚焦超声(HIFU)制备抗原致敏树突状细胞(DC)及其诱导细胞毒性T淋巴细胞(CTL)的杀伤效应。方法应用小鼠重组GM-CSF和IL-4培养小鼠骨髓DC,用HIFU制备CT26肿瘤细胞抗原致敏DC疫苗,用3H-TdR法检测T细胞增殖反应的能力,标准的4h51Cr释放测定法检测CTL杀伤活性。结果HIFU组、肿瘤细胞冻融组、肿瘤上清组和PBS组的DC在体外均可以诱导CTL增殖,但HIFU组、肿瘤细胞冻融组的DC体外诱导CTL增殖能力明显高于肿瘤上清组和PBS组(P<0.05)。HIFU组和肿瘤细胞冻融组DC体外诱导的CTL对CT26结肠癌均有明显的细胞毒性作用,与肿瘤上清组DC和PBS组比较差异有统计学意义(P<0.05),HIFU组体外诱导的CTL杀伤活性与肿瘤细胞冻融组比较差异无统计学意义(P>0.05)。结论HIFU制备抗原致敏DC可以诱导CTL大量增殖,并能诱导出杀伤效应较强的CTL。
Objective To investigate the killing effect of antigen-responsive dendritic cells (DCs) and cytotoxic T lymphocytes (CTLs) produced by high intensity focused ultrasound (HIFU). Methods Murine bone marrow DCs were cultured with recombinant mouse GM-CSF and IL-4. DC vaccines primed with CT26 tumor cell antigen were prepared using HIFU. The ability of T-cell proliferation was assayed by 3H-TdR. The standard 4h51Cr release assay CTL killing activity. Results The proliferation of CTLs was induced in HIFU, tumor cell freezing and thawing groups, tumor supernatants and PBS groups in vitro. However, the proliferation of CTL induced by DC in vitro in HIFU group and tumor cell freezing group was significantly higher than that in tumor supernatant group And PBS group (P <0.05). CTLs induced by DCs in HIFU group and frozen / thawed cells group had obvious cytotoxicity on CT26 colon cancer, which was significantly different from that in DC and PBS groups (P <0.05), and induced by HIFU in vitro CTL cytotoxic activity was not significantly different from tumor cell freezing and thawing group (P> 0.05). Conclusion The preparation of antigen-primed DCs by HIFU can induce the proliferation of CTLs and induce CTLs with high cytotoxicity.