Li Qi-Ling, Wang Yun-Ping, Gao Shang-Feng, Ma Jun, Feng Cai-Xia, Wang Yue-Ling
Department of Obstetrics and Gynecology, First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China.
Zhonghua Fu Chan Ke Za Zhi. 2009 Nov;44(11):856-60.
To explore the mechanisms and effects of adoptive immunotherapy with ovarian cancer vaccine modified by GM-CSF gene which was used after immunologic reconstitution during lymphopenia induced by chemotherapy.
Lymphopenia was induced by chemotherapy with cyclophosphamide. The immune reconstituted model was built in rats. The tumor vaccine draining lymph nodes were harvested after the ovarian cancer cells NUTU-19 modified by GM-CSF gene were injected. The effector T cells (T(E)) were got after being stimulated and amplified. Enzyme-linked immunosorbent assay was used to detect the level of interleukin (IL)-2 and IL-4 secreted by T(E). Intracellular cytokine staining was used to determine frequency of tumor-specific T(E). Fluorescence-activated cell sorting (FACS) was used to detect the special cytotoxicity of T(E) killing target cells. The survival period of rats bearing pre-established abdominal ovariam carcinoma after being adoptively transferred by T(E) was observed.
Compared with those in control group, the significant higher levels IL-2 [(65.7 +/- 4.0) pg/ml] and lower levels IL-4 [(277 +/- 49) pg/ml] were observed in chemotherapy-immune reconstitution-vaccine immunization group. The amount of CD(4)(+) T cells secreting interferon-gamma (13.0 +/- 2.1)% were also significantly increased. The rate of the special cytotoxicity of killing T cells (86.5 +/- 1.1)% was markedly improved. The survival period of rats (110 +/- 16) days was increased in chemotherapy-immune reconstitution-vaccine immunization group.
The combined immunotherapy of chemotherapy-immune reconstitution-tumor vaccine immunotherapy may increase the frequency and function of specific tumor T(E). The specific cytotoxicity is increased and the weak reaction of T(E) to tumor is improved, which showed that this therapy can enhance immune reaction.
探讨化疗诱导淋巴细胞减少期间免疫重建后使用GM-CSF基因修饰的卵巢癌疫苗进行过继性免疫治疗的机制及效果。
用环磷酰胺化疗诱导淋巴细胞减少，建立大鼠免疫重建模型。注射GM-CSF基因修饰的卵巢癌细胞NUTU-19后，收集肿瘤疫苗引流淋巴结。刺激扩增后获得效应T细胞（T(E)）。采用酶联免疫吸附测定法检测T(E)分泌的白细胞介素（IL）-2和IL-4水平。采用细胞内细胞因子染色法测定肿瘤特异性T(E)的频率。采用荧光激活细胞分选术（FACS）检测T(E)杀伤靶细胞的特异性细胞毒性。观察预先建立腹部卵巢癌的大鼠经T(E)过继转移后的生存期。
化疗-免疫重建-疫苗免疫组与对照组相比，IL-2水平显著升高[（65.7±4.0）pg/ml]，IL-4水平显著降低[（277±49）pg/ml]。分泌γ干扰素的CD(4)(+)T细胞数量（13.0±2.1）%也显著增加。杀伤性T细胞的特异性细胞毒性率（86.5±1.1）%明显提高。化疗-免疫重建-疫苗免疫组大鼠的生存期延长至（110±16）天。
化疗-免疫重建-肿瘤疫苗免疫联合治疗可能增加特异性肿瘤T(E)的频率和功能。特异性细胞毒性增强，T(E)对肿瘤的微弱反应得到改善，表明该疗法可增强免疫反应。
