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目的探讨HCA587蛋白疫苗对荷瘤小鼠的抗肿瘤效应并揭示抗肿瘤效应的免疫细胞亚群。方法在C57BL/6小鼠左侧胁腹部皮下接种肿瘤细胞建立肿瘤模型,给予HCA587蛋白疫苗等免疫策略治疗2次,观察抗肿瘤效果。用苏木精-伊红(HE)染色法分析疫苗治疗后肿瘤组织中淋巴细胞的浸润。用SDS-PAGE电泳分析自行制备的抗CD4和CD8分子单抗的相对分子质量和纯度,并用流式细胞术对其体内阻断效果进行鉴定。将上述单抗腹腔内注射入小鼠体内,以实现免疫细胞在体清除。结果与单一蛋白治疗组、佐剂对照组相比,HCA587蛋白疫苗具有显著的抗肿瘤作用(P<0.05)。在HE染色中,HCA587蛋白疫苗治疗后肿瘤局部可见大量的免疫细胞浸润。经流式细胞术鉴定,自行制备、纯化的抗CD4和CD8分子单抗在体能完全清除相应的CD4+T和CD8+T细胞亚群。当在体清除CD4+T细胞后,HCA587蛋白疫苗的抗肿瘤效应完全消失,而CD8+T细胞的清除不影响其抗肿瘤效应。结论 HCA587蛋白疫苗具有显著的抗肿瘤效应,其诱导的CD4+T细胞在抗肿瘤效应中起着至关重要的作用。
Objective To investigate the antitumor effect of HCA587 protein vaccine on tumor-bearing mice and to reveal the anti-tumor immune cell subsets. Methods C57BL / 6 mice were inoculated subcutaneously in the flank of the left flank to establish a tumor model. The mice were immunized with HCA587 protein vaccine for 2 times. The antitumor effect was observed. Lymphocyte infiltration in tumor tissue after vaccination was analyzed by hematoxylin-eosin (HE) staining. The relative molecular mass and purity of anti-CD4 and CD8 monoclonal antibodies prepared by themselves were analyzed by SDS-PAGE electrophoresis, and their blocking effect was identified by flow cytometry. The above monoclonal antibody intraperitoneal injection into mice in order to achieve immune cells in vivo clearance. Results HCA587 protein vaccine had significant anti-tumor effect (P <0.05) compared with single protein treatment group and adjuvant control group. In HE staining, a large amount of immune cell infiltration was observed in the tumor area after treatment with HCA587 protein vaccine. By flow cytometry, self-prepared and purified anti-CD4 and CD8 mAb completely eliminated the corresponding CD4 + T and CD8 + T cell subsets in vivo. The antitumor effect of the HCA587 protein vaccine disappeared completely after CD4 + T cells were cleared in vivo, whereas clearance of CD8 + T cells did not affect their anti-tumor effect. Conclusion HCA587 protein vaccine has significant anti-tumor effect, and its induced CD4 + T cells play a crucial role in anti-tumor effect.