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目的:荷瘤宿主淋巴细胞体外活化后用于免疫重建,辅以瘤苗免疫,体外检测免疫学指标变化情况。方法:以环磷酰胺处理雌性近交系荷瘤Fischer344大鼠,建立化疗后淋巴细胞减少(LP)模型;用同系荷瘤大鼠淋巴细胞体外活化扩增后对LP大鼠进行免疫重建,免疫重建24h内,用GM-CSF修饰的大鼠卵巢癌细胞株NuTu-19免疫,8～10d后采集肿瘤疫苗部位引流淋巴结(TVDLN),以流式细胞仪(FACS)分析细胞表型,检测MHC-II和CD86验证树突状细胞(DC)的增殖能力,分析FITC-CD4和PE-CD8的比例以此反映T细胞的激活情况,未成熟DC(iDC)对葡聚糖(FITC-Dextran)的吞噬能力,反映DC对抗原的处理和提呈功能。结果:荷瘤宿主淋巴细胞体外活化组TVDLN中,DC和活化T细胞的频数显著增高,iDC的能力显著增强。结论:荷瘤宿主淋巴细胞体外活化后于淋巴细胞减少期,进行免疫重建,辅以瘤苗治疗,可选择性扩增、活化抗原特异T细胞和DC,增强抗肿瘤免疫反应。
OBJECTIVE: Tumor-bearing host lymphocytes activated in vitro for immune reconstitution, supplemented with vaccine-immunized, in vitro changes in immunological indicators. Methods: The female inbred tumor-bearing Fischer344 rats were treated with cyclophosphamide to establish a model of lymphopenia (LP) after chemotherapy. Lymphocytes were revascularized in vitro and re-immunized with LP Immunohistochemistry was used to immunize rat ovarian cancer cell line NuTu-19 modified with GM-CSF within 24h. Eight days after transplantation, the tumor-draining lymph nodes (TVDLN) were collected. The cell phenotypes were analyzed by flow cytometry (FACS) -II and CD86 to verify the proliferation of dendritic cells (DCs). The proportion of FITC-CD4 and PE-CD8 was analyzed to reflect the activation of T cells. The effects of imDC on dextran (FITC-Dextran) Phagocytosis, reflecting the DC on antigen processing and presentation functions. Results: The frequency of DCs and activated T cells was significantly increased in TVDLN of tumor-bearing host lymphocytes activated in vitro, and the ability of iDCs was significantly enhanced. CONCLUSION: Tumor-bearing host lymphocytes activated in vitro and then re-immunized during lymphopenia with adjuvant tumor vaccine can selectively amplify and activate antigen-specific T cells and DC to enhance anti-tumor immune response.