热休克蛋白Hsp 70激活树突状细胞治疗肺癌

Experimental Study of Curative Effect of Dendritic Cells Activated by Heat Shock Protein 70 on Lung Cancer in Nude Mice

目的利用热休克蛋白Hsp 70作为佐剂增强树突状细胞(DCs)递呈肿瘤抗原的能力并提高细胞毒T淋巴细胞(CTLs)对肺癌细胞的杀伤活性。方法外周血单个核细胞体外经GM-CSF和IL-4诱导产生树突状细胞,负载A549肺癌细胞裂解物的同时加入Hsp 70,诱导的自体CTLs用细胞毒试验和ELISA测定杀伤活性和细胞因子的分泌。同时建立荷瘤裸鼠模型,不同分组裸鼠一次或多次皮下注射CTLs。结果A 549冻融抗原+Hsp 70显著增强CTLs的增殖能力,诱导的CTLs对A 549细胞产生特异性杀伤并能抑制荷瘤裸鼠的肿瘤生长。结论DCs能有效呈递肺癌冻融抗原,诱导产生抗原特异性CTLs。在抗原致敏阶段加入Hsp 70能使CTLs杀伤活性进一步增强,提示Hsp 70在以DCs为基础的肺癌疫苗和过继免疫治疗中具有广阔前景。

Objective To observe the curative effect of dendritic cells（DCs） activated by heat shock protein 70（Hsp 70） on lung cancers. Methods Autologous DCs were derived from peripheral blood mononuclear cells （PBMC） in healthful men by inducement in vitro with granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin-4（IL- 4）. The dendritic cells were loaded with A549 tumor cell freeze-thaw lysate and at the same time Hsp 70 as an immune adjuvant was added. The specific groupings induced generation of tumor specific Cytotoxic T lymph cells （CTLs）. Killing activity and cytokine release of the CTLs and the population of CTLs were measured by cytotoxic assay and ELISA. The lung cancer model was established in the nude mice. The Hsp70-antigen/DCs induced by CTLs were subcutaneously injected （2 x 107/mouse） 1 or 3 times per week after the A 549 tumor cells were inoculated. The therapeutic effect was observed. Results DCs pulsed with A 549 tumor cell lysate enhanced the CTLs. The cytotoxicity of specific CTLs against A 549 lung cancer cells was significantly enhanced and the survival time of tumor-bearing mice was prolonged when the specific CTLs were injected subcutaneously repeatedly. Conclusion The ability of Hsp 70 to activate DCs indicates its potential as a novel adjuvant for peptide immunizations; The Hsp 70 antigen peptide hybrid may serve as a more effective vaccine for the control of lung cancer.