巨噬细胞瘤苗免疫调节作用的研究

Observation on the immunoregulation function of the macrophage tumor vaccine

目的 观察巨噬细胞肿瘤疫苗对细胞毒性T细胞（CTL）反应及Th1／Th2型细胞因子分泌的调节作用。方法 分别采用MTT法和比色法检测肿瘤细胞杀伤率和清液上乳酸脱氢酶（LDH）含量，采用Western印记法与ELISA法检测脾细胞内及分泌入培养上清的IL-2、IL-4、IL-10及IFN-γ等细胞因子。结果 巨噬细胞肿瘤疫苗接种组的淋巴细胞肿瘤细胞杀伤率与培养上清液LDH水平分别为（36．70±21．02）％和（0．29±0．15）U／ml，明显高于热灭活肿瘤细胞接种组、石蜡诱导的巨噬细胞接种组（P值均〈0．05）。肿瘤细胞冻融物刺激72h后，各组免疫小鼠的脾细胞内均可检测到IL-2、IL-4、IL-10及IFN-γ等细胞因子；同时巨噬细胞肿瘤疫苗接种组脾细胞培养上清中，IL-2、IL-4、IL-10及IFN-γ含量分别为（7．97±3．15）pg／ml、（0．44±0．11）pg／ml、（1．83±0．85）pg／ml和（9．16±4．64）pg／ml，IL-2、IFN-γ的水平均高于热灭活肿瘤细胞接种组和石蜡诱导的巨噬细胞接种组（P值均〈0．05）。结论 巨噬细胞肿瘤疫苗可诱导机体产生特异性抗肿瘤反应，能够促进Th1型细胞因子IL-2、IFN-γ的分泌。

Objective To investgate the effect of macrophage tumor vaccine on the regulation of CTL response and Th1/Th2 type cytokines production. Methods The tumor cell injury rate and the level of LDH in culture supernatant were detected with MrIT method and colorimetric assay. Cytokine IL-2, IL-4, IL-10 and IFN-γ, expressed in spleen cells and released into culture supernatant were detected with Western bolting and ELISA, respectively. Results The tumor injury rate and the level of LDH in culture supernatant of macrophage tumor vaccine immunized group were （36. 70±21.02）% and （0. 29±0. 15） U/ml, respectively. Both are significantly higher than that of the group immunized with heat inactivated tumor cells and the group immunized with liquid paraffin induced macrophages （P 〈 0. 05 ）. After stimulating 72 hours with the freeze thawing tumor cells, spleen cells of every group expressed cytokine IL-2, IL-4, IL-10 and IFN-γ. Meanwhile, levels of IL-2, IL-4, IL-10 and IFN-γ in culture supernatant of macrophage tumor vaccine immunized group were （7. 97±3. 15 ） pg/ ml, （0. 44±0. 11 ） pg/ml, （ 1.83± 0. 85 ） pg/ml and （9. 16±4. 64） pg/ml, respectively. All of them obviously higher than that of the group of immunized with heat inactivated tumor cells and the group immunized with liquid paraffin induced macrophages（ P 〈 0.05 ）. Conclusion Macrophage tumor vaccine could elicit specific anti-tumor immune response and promote spleen cells to secrete Thl type eytokines such as IL-2 and IFN-γ.