He Xiangfeng, Shi Wen, Zhao Fengshu, Wang Jianhong, Xu Xiaohong, Tan Qinghe, Sun Yongqiang, Chen Dengyu, Dou Jun
Department of Medicine, Nantong Cancer Hospital Affiliated to Nantong University, Nantong 226361, China.
Email:
Zhonghua Zhong Liu Za Zhi. 2014 Apr;36(4):245-9.
To investigate the effect and mechanism of B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine on pulmonary metastasis in mouse model of melanoma.
Twelve 8-week old female C57BL/6 mice were used in this study. The mice were injected with wild-type B16F10 cells through tail vein after immunization with B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine, and the pulmonary metastasis was observed. The CD4(+) and CD8(+) T cells were isolated by magnetic activated cell sorting, and then used for the detection of CFSE/7-AAD cytotoxicity by flow cytometry. Serum from the mice immunized with tumor-cell vaccine was used to detect IFN-γ expression by ELISA. The expression of TGF-β2, ZEB1, E-cadherin, and N-cadherin of tumor tissues was detected by RT-PCR and immunofluorescence, respectively.
The mice vaccinated with B16F10-ESAT-6-gpi/IL-21 had significantly fewer nodules in the lung and lower lung weight [(285.8 ± 19.01) mg vs. (406.3 ± 27.12) mg], with lower levels of TGF-β2, ZEB1 and N-cadherin proteins but higher level of E-cadherin protein within the tumor tissue, as compared with the control mice. Meanwhile, the immunized mice had significantly increased CD8(+) T cell killing activity [(42.62 ± 3.465)% vs. (22.29 ± 1.804)%] and IFN-γ expression level [(55.200 ± 7.173) pg/ml vs. (6.435 ± 1.339) pg/ml] over the control mice.
The B16F10-ESAT-6-gpi/IL-21 vaccine can inhibit the metastasis of melanoma in the lung in vaccinated melanoma-bearing mice. This inhibitory effect is associated with CD8(+) T cell immune response and a higher level of IFN-γ, which may influence on the mesenchymal-epithelial transition of tumor cells.
探讨B16F10-ESAT-6-gpi/IL-21肿瘤细胞疫苗对黑色素瘤小鼠模型肺转移的影响及其机制。
本研究选用12只8周龄雌性C57BL/6小鼠。用B16F10-ESAT-6-gpi/IL-21肿瘤细胞疫苗免疫小鼠后，经尾静脉注射野生型B16F10细胞，观察肺转移情况。通过磁珠分选法分离CD4(+)和CD8(+) T细胞，然后用流式细胞术检测CFSE/7-AAD细胞毒性。用肿瘤细胞疫苗免疫小鼠的血清通过ELISA法检测IFN-γ表达。分别用RT-PCR和免疫荧光法检测肿瘤组织中TGF-β2、ZEB1、E-钙黏蛋白和N-钙黏蛋白的表达。
与对照小鼠相比，接种B16F10-ESAT-6-gpi/IL-21的小鼠肺内结节明显减少，肺重量降低[（285.8±19.01）mg对（406.3±27.12）mg]，肿瘤组织内TGF-β2、ZEB1和N-钙黏蛋白水平较低，但E-钙黏蛋白水平较高。同时，免疫小鼠的CD8(+) T细胞杀伤活性[（42.62±3.465）%对（22.29±1.804）%]和IFN-γ表达水平[（55.200±7.173）pg/ml对（6.435±1.339）pg/ml]均显著高于对照小鼠。
B16F10-ESAT-6-gpi/IL-21疫苗可抑制接种黑色素瘤小鼠的黑色素瘤肺转移。这种抑制作用与CD8(+) T细胞免疫反应及较高水平的IFN-γ有关，可能影响肿瘤细胞的间质-上皮转化。
