Gal-1单抗增强γδT细胞对宫颈癌细胞的杀伤作用

Gal-1 monoclonal antibody enhances cytotoxicity of γδT cells against cervical carcinoma cells

目的:探讨γδT细胞联合半乳糖凝集素1(galectin-1,Gal-1)单抗对人宫颈癌细胞的杀伤作用。方法:从人宫颈癌组织中分离肿瘤浸润淋巴细胞(tumor infiltrating lymphocytes,TILs),并通过固相单抗包被法在体外扩增获得γδT细胞,流式细胞术检测其纯度;Western blotting和ELISA法检测宫颈癌SiHa、HeLa细胞及其上清中Gal-1的表达,乳酸脱氢酶(lactate de-hydrogenase,LDH)释放实验检测γδT细胞联合Gal-1单抗对宫颈癌SiHa、HeLa细胞的杀伤作用。皮下注射SiHa细胞制备荷瘤小鼠模型,随机分为对照组、同型对照组(IgG1组)、γδT细胞组(γδT组)、Gal-1单抗组(Gal-1mAb组)及γδT细胞联合Gal-1单抗组(γδT+Gal-1 mAb组),观察各组荷瘤小鼠移植瘤生长的情况。结果:固相单抗包被法体外扩增后TCRγδ阳性γδT细胞比例达(91.2±1.2)%,SiHa、HeLa细胞及其上清中Gal-1高表达。与对照组相比,Gal-1 mAb组能够增强γδT细胞体外杀伤SiHa细胞[(68.1±3.0)%vs(48.7±3.8)%,P<0.05]和HeLa细胞[(79.4±5.6)%vs(48.3±6.5)%,P<0.05]的效率。裸鼠荷瘤30 d,γδT+Gal-1 mAb组的移植瘤体积明显小于Gal-1单抗组[(31.3±9.1)vs(199.6±41.2)mm3,P<0.01]和γδT细胞组[(31.3±9.1)vs(85.6±45.1)mm3,P<0.05]。结论:Gal-1单抗能增强γδT细胞对宫颈癌细胞的杀伤作用。

Objective：To investigate the cytotoxicity of γδT cells combined with galectin-1 （Gal-1） monoclonal antibody against human cervical carcinoma cells. Methods： γδT cells were expanded in vitro using the solid-phase antibody coated method from tumor infiltrating lymphocyte （TILs） of the human cervical cancer specimens. The purity of γδT cells was measured by flow cytometry. The expressions of Gal-1 in cervical carcinoma SiHa and HeLa cells and in cell supernatants were detected by Western blotting and ELISA, respectively. The cytotoxicity of γδT cells combined with Gal-1 mAb against the human cervical cancer SiHa and HeLa cells was measured by lactate dehydrogenase （LDH） release assay. Tumor-bearing mouse model was established by subcutaneous injection of SiHa cells. Tumor-bearing mice were randomly divided into a control group （SiHa group）, an isotype control group （SiHa＋mouse IgG1 group）, a γδT cell group （SiHa＋γδT group）, a Gal-1 mAb group （SiHa＋γδT＋Gal-1 mAb group） and a γδT cell ＋ Gal-1 mAb group （γδT cell＋Gal-1 mAb group）. The tumor growth was observed in different groups. Results： The percentage of TCRγδ positive T cells expanded using the solid-phase antibody coated method in vitro was （91.2±1.2） %. Gal-1 was over-expressed in SiHa, HeLa cells and cell supernatants. Compared with the control group, the Gal-1 mAb group showed an increasing cytotoxicity efficiency on SiHa cells （／[68.1±3.0／] % vs ／[48.7±3.8／]%, P〈005） and HeLa cells （／[79.4±5.6／]% vs ／[48.3±6.5／]%, P〈0.05）. The volume of implanted tumors in the γδT ＋Gal-1 mAb group was significantly smaller than that in the Gal-1 mAb group （／[31.3±9.1／] vs ／[199.6±41.2／] mm3, P〈0.01） and the γδT cell group （／[31.3±9.1／] vs ／[85.6±45.1／] mm3, P〈0.05） 30 days after tumor implantation in nude mice. Conclusion： Gal-1 mAb antibody can boost up the cytotoxicity of γδT cells against the human cervical carcinoma cells.