热休克蛋白110-人表皮生长因子受体2/神经胶质瘤细胞内区免疫的BALB/c小鼠脾脏淋巴细胞的细胞毒性活性

[Cytotoxic activity of spleen lymphocytes in BALB/c mice immunized by HSP110-HER2/neu ICD].

作者信息

Han Dong, Xu Huang, Yan Wei-qun

机构信息

Department of Biochemistry, Jiaxing College, Jiaxing, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2012 Jan;34(1):11-4.

PMID:22490848

Abstract

OBJECTIVE

To explore the cytotoxic responses of spleen T lymphocytes (CTL) in BALB/c mice induced by recombinant HSP110-HER2/neu ICD complex.

METHODS

Tumor-bearing mouse model was immunized by HSP110-HER2/neu ICD complex. The IFN-γ level secreted by activated spleen T lymphocytes was detected by enzyme linked immunospot assay (ELISPOT). The corresponding CTL activity was measured by granzyme release assay.

RESULTS

The BALB/c mouse model of human mammary tumor highly expressing HER2/neu was established. HSP110-HER2/neu ICD complex immunization led to a significantly higher level of INF-γ than that in HSP110-P(789-797) immunized and HER2/neu ICD immunized mice. HSP110-HER2/neu ICD complex immunized animals also show significant CTL activity. The results of immunohistochemical staining showed that the number of blue spots in the PBS group was 4.57 ± 1.33, HSP110 group 6.83 ± 2.08, HER2/neu ICD group 16.17 ± 2.86, HSP110-P(789-797) group 43.67 ± 4.78, and SP110-HER2/neu ICD group 76.51 ± 8.17. The number of IFN-γ-secreting spleen lymphocytes in the HSP110-HER2/neu ICD group was significantly higher than that in the HSP110-P(789-797) group, and that of HSP110-P(789-797) group was significantly higher than that of HER2/neu ICD group (P < 0.01). The target cell-killing rate of the PBS group was (8.15 ± 1.27)%, HSP110 group (9.51 ± 1.51)%, HER2/neu ICD group (14.03 ± 2.45)%, HSP110-P(789-797) group (25.99 ± 3.04)% and HSP110-HER2/neu ICD group (38.15 ± 3.95)% (all P < 0.01).

CONCLUSIONS

HSP110-HER2/neu ICD complex can promote the proliferation and maturation of T lymphocytes into CTLs, and might be used as anti-tumor vaccine to induce potent cytotoxic T lymophocyte immunoresponse against specific tumor cells.

摘要

目的

探讨重组热休克蛋白110-人表皮生长因子受体2/神经胶质瘤抑制因子（HSP110-HER2/neu ICD）复合物诱导BALB/c小鼠脾脏T淋巴细胞（CTL）的细胞毒性反应。

方法

用HSP110-HER2/neu ICD复合物免疫荷瘤小鼠模型。采用酶联免疫斑点法（ELISPOT）检测活化的脾脏T淋巴细胞分泌的γ干扰素（IFN-γ）水平。通过颗粒酶释放试验测定相应的CTL活性。

结果

建立了高表达HER2/neu的人乳腺肿瘤BALB/c小鼠模型。HSP110-HER2/neu ICD复合物免疫导致INF-γ水平显著高于HSP110-P(789-797)免疫组和HER2/neu ICD免疫组小鼠。HSP110-HER2/neu ICD复合物免疫的动物也表现出显著的CTL活性。免疫组化染色结果显示，磷酸盐缓冲液（PBS）组蓝斑数为4.57±1.33，HSP110组为6.83±2.08，HER2/neu ICD组为16.17±2.86，HSP110-P(789-797)组为43.67±4.78，SP110-HER2/neu ICD组为76.51±8.17。HSP110-HER2/neu ICD组分泌IFN-γ的脾脏淋巴细胞数量显著高于HSP110-P(789-797)组，HSP110-P(789-797)组显著高于HER2/neu ICD组（P<0.01）。PBS组的靶细胞杀伤率为(8.15±1.27)%，HSP110组为(9.51±1.51)%，HER2/neu ICD组为(14.03±2.45)%，HSP110-P(789-797)组为(25.99±3.04)%，HSP110-HER2/neu ICD组为(38.15±3.95)%（均P< 0.01）。