荷瘤宿主淋巴细胞体外活化后用于免疫重建免疫学指标变化

Evaluation of immune reconstitution with in vitro-activated T cells from a tumor-bearing host

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。

AIM：To evaluate the reconstituted immune system with in vitro-activated T cells from tumor-bearing rats coupled with ovarian cancer vaccination. METHODS： Fischer 344 female rats were injected with cyclophosphamide （CY） as a lymphopenia （LP） model. The immune systems of the rats were reconstituted with in vitro-activated T cells from the same individuals. GM-CSF-modified ovarian cancer cells lines （NuTu-19） were injected within 24 h after immune reconstitution. The tumor vaccine draining lymph nodes （TVDLN） were harvested 8-10 days after vaccination and analyzed by FACS. The proliferative capacity of dendritic cells （DCs） was measured by the levels of MHC-II and CD86 molecules. The activation of T cells was monitored by the percentage of FITC-CD4 and PE-CD8 cells. The biological function of DCs such as processing and presenting antigens was assayed by immature DCs’ phagocytosis of FITC-Dextran. RESULTS： Immune reconstitution with in vitro-activated T cells produced significantly more DCs, T cells and functionally enhanced immature DCs out of TVDLN. CONCLUSION： Reconstituting immune system with in vitro-activated T cells from a tumor-bearing host coupled with ovarian cancer vaccination during lymphocytopenia may selectively expand and activate particular T cells and DCs, leading to augmentation of anti-tumor immunity.