NY-ESO-1/HSP65融合蛋白腹腔免疫小鼠的免疫效果

目的  观察NY-ESO-1/热休克蛋白65（heat shock protein 65，HSP65）融合蛋白（NY-H）免疫BALB/c小鼠诱导的体液和细胞免疫应答及其抗肿瘤作用。 方法  将BALB/c小鼠按简单随机方法分组，分别用 NY-H、NY-ESO-1与HSP65混合（NY+H）、NY-ESO-1、HSP65和PBS腹腔免疫4次，间隔2周。每次免疫后第14天，对小鼠尾静脉采血，检测血清抗体滴度。末次免疫后第7天，取小鼠脾细胞，检测细胞因子、淋巴细胞增殖水平、CD4+和CD8+ T细胞的百分比及对肿瘤细胞的特异性杀伤作用，采用t检验或2检验对结果进行比较。结果 NY-H组小鼠在第1次免疫后就产生了较高水平的抗体（吸光度值为0.841±0.059），与NY+H组（0.333±0.077）和NY-ESO-1组（0.275±0.183）相比差异有统计学意义（t=12.753，P<0.01；t=7.204，P<0.01）。NY-H组小鼠的IL-2和IFN-γ分泌能力均高于其他各组，并且CD4+、CD8+ T细胞数量大幅增高，细胞增殖率（38.00%±3.84%）明显高于NY+H组（12.90%±0.65%）和NY-ESO-1组（10.90%±1.44%）（x=830.1，P<0.01；x²=994.0，P<0.01)。小鼠免疫NY-H后，脾淋巴细胞能特异性杀伤表达NY-ESO-1的小鼠B16肿瘤细胞。 结论  NY-H能诱导小鼠产生较强的体液和细胞免疫应答，且具有良好的抗肿瘤作用。

Immunological effect of intraperitoneal immunization with NY-ESO-1/HSP65 fusion protein in BALB/c mice

Objective  To evaluate humoral and cellular immune responses to NY-ESO-1/ heat shock protein 65 (HSP65) fusion protein (NY-H) and its anti-tumor effect in mice. Methods BALB/c mice were simply randomized into 5 groups and intraperitoneally injected with NY-H, NY-ESO-1 mixed with HSP65 (NY+H), NY-ESO-1, HSP65 and PBS, respectively, every 2 weeks for 4 times. Antibody titers were detected at 14 d after each immunization. Secretion of cytokines, proliferation level of spleen lymphocytes, percentages of CD4+and CD8+ T cells, and ability of T cells to lyse tumor cells were evaluated at 7 d after the last immunization. The results were compared using t-test or 2-test.  Results   NY-H elicited strong and prompt anti-NY-ESO-1 antibody response with the absorbance value of 0.841±0.059, which was significantly higher than those in NY+H (0.333±0.077) and NY-ESO-1 groups (0.275±0.183) (t=12.753, P<0.01; t=7.204, P<0.01). The ability of IL-2 and IFN-γ secretion in NY-H group was the strongest. The percentage of CD4+ and CD8+ T cells in NY-H group (38.00%±3.84%) was significantly higher than those in NY+H group (12.90%±0.65%) and NY-ESO-1 group (10.90%±1.44%) (2=830.1, P<0.01; 2=994.0, P<0.01), suggesting that NY-H elicited cellular immune responses effectively. Spleen lymphocytes isolated from NY-H-immunized mice were shown to specifically lyse NY-ESO-1-expressing B16 cells.  Conclusions The fusion protein NY-H could induce strong humoral and cellular immune responses and has good anti-tumor effect.