肿瘤浸润淋巴细胞(tumor-infiltrating lymphocytes,TILs)介导的过继性免疫治疗已在多种实体瘤中取得良好疗效。然而,传统TILs体外扩增方法效率低,难以达到治疗级别的细胞数量和高肿瘤杀伤活性要求。为了研究3D肿瘤模型对TILs体外激活...肿瘤浸润淋巴细胞(tumor-infiltrating lymphocytes,TILs)介导的过继性免疫治疗已在多种实体瘤中取得良好疗效。然而,传统TILs体外扩增方法效率低,难以达到治疗级别的细胞数量和高肿瘤杀伤活性要求。为了研究3D肿瘤模型对TILs体外激活扩增及肿瘤杀伤活性的影响,为TILs体外扩增提供新策略,本研究从肺癌患者手术样本中获取TILs和原代肿瘤细胞,对比观察2D及3D培养条件下肺癌细胞系NCI-H1975及原代肺癌细胞对TILs激活扩增及抗肿瘤作用的影响;向3D培养原代肿瘤~+TILs中添加程序性死亡受体1(programmed cell death protein 1,PD-1)抗体,验证PD-1抗体对该体系中TILs浸润杀伤肿瘤细胞的影响。结果表明,相比于2D培养,3D培养的H1975肺癌细胞系可使TILs在一定时间内显著扩增,并提升TILs中CD3~+/CD8~+细胞的比例(61.6%);3D培养的原代肿瘤球也可刺激增加CD3~+/CD8~+TILs细胞的比例(45.5%,54.4%),诱导肿瘤上皮细胞的凋亡,使其存活率降至16.7%;进一步研究表明,PD-1抗体的引入促进3D原代肿瘤球共培养介导的TILs的增殖、提高CD3~+/CD8~+细胞的比例(50.9%,57.0%),并显著提高其抗肿瘤效力(肿瘤上皮细胞整体存活率降至9.36%)。综上所述,3D肿瘤球显著增强TILs细胞的体外激活增殖及抗肿瘤能力,且PD-1抗体进一步促进3D肿瘤球介导的TILs细胞增殖及其杀伤肿瘤的效力。展开更多
Establishing Epstein-Barr virus (EBV)-specific cytolytic T lymphocytes (EBV-CTLs) from peripheral blood mononuclear cells (PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including H...Establishing Epstein-Barr virus (EBV)-specific cytolytic T lymphocytes (EBV-CTLs) from peripheral blood mononuclear cells (PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma (NPC). In the current study,we performed ex vivo expansion of tumor-infiltrating lymphocytes (TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody (OKT3), recombinant human interleukin (IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+T cells, a variable percentage of CD3+CD8+and CD3+CD4+T cells, and less than 10% of CD3-CD16+natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.展开更多
文摘肿瘤浸润淋巴细胞(tumor-infiltrating lymphocytes,TILs)介导的过继性免疫治疗已在多种实体瘤中取得良好疗效。然而,传统TILs体外扩增方法效率低,难以达到治疗级别的细胞数量和高肿瘤杀伤活性要求。为了研究3D肿瘤模型对TILs体外激活扩增及肿瘤杀伤活性的影响,为TILs体外扩增提供新策略,本研究从肺癌患者手术样本中获取TILs和原代肿瘤细胞,对比观察2D及3D培养条件下肺癌细胞系NCI-H1975及原代肺癌细胞对TILs激活扩增及抗肿瘤作用的影响;向3D培养原代肿瘤~+TILs中添加程序性死亡受体1(programmed cell death protein 1,PD-1)抗体,验证PD-1抗体对该体系中TILs浸润杀伤肿瘤细胞的影响。结果表明,相比于2D培养,3D培养的H1975肺癌细胞系可使TILs在一定时间内显著扩增,并提升TILs中CD3~+/CD8~+细胞的比例(61.6%);3D培养的原代肿瘤球也可刺激增加CD3~+/CD8~+TILs细胞的比例(45.5%,54.4%),诱导肿瘤上皮细胞的凋亡,使其存活率降至16.7%;进一步研究表明,PD-1抗体的引入促进3D原代肿瘤球共培养介导的TILs的增殖、提高CD3~+/CD8~+细胞的比例(50.9%,57.0%),并显著提高其抗肿瘤效力(肿瘤上皮细胞整体存活率降至9.36%)。综上所述,3D肿瘤球显著增强TILs细胞的体外激活增殖及抗肿瘤能力,且PD-1抗体进一步促进3D肿瘤球介导的TILs细胞增殖及其杀伤肿瘤的效力。
基金supported by grants from the National Natural Science Foundation of China (No.224-30872981)Guangdong Province Natural Science Foundation (No.10151008901000156)
文摘Establishing Epstein-Barr virus (EBV)-specific cytolytic T lymphocytes (EBV-CTLs) from peripheral blood mononuclear cells (PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma (NPC). In the current study,we performed ex vivo expansion of tumor-infiltrating lymphocytes (TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody (OKT3), recombinant human interleukin (IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+T cells, a variable percentage of CD3+CD8+and CD3+CD4+T cells, and less than 10% of CD3-CD16+natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.
