Human dendritic cells transfected with amplified MUC1 mRNA stimulate cytotoxic T lymphocyte responses against pancreatic cancer in vitro
Human dendritic cells transfected with amplified MUC1 mRNA stimulate cytotoxic T lymphocyte responses against pancreatic cancer in vitro
复制标题
DOI：
10.1111/j.1440-1746.2011.06778.x
复制
发表时间：
2011-10-01
影响因子：
4.1
通讯作者：
Guo, Xiao-Zhong
中科院分区：
文献类型：
--
作者：
Chen, Jiang;Li, Hong-Yu;Guo, Xiao-Zhong
背景和目的：粘蛋白（MUC）1是一种上皮细胞糖蛋白，在包括胰腺癌（PC）在内的许多腺癌中异常过度表达，为免疫治疗提供了理想的肿瘤相关抗原和靶点。在本研究中，我们研究了转染扩增的MUC1 mRNA的树突状细胞（DC）诱导的细胞毒性T淋巴细胞（CTL）是否能够在体外对PC产生反应。方法：使用优化设置的电穿孔将编码MUC1的扩增的mRNA转染到DC中，并通过实时定量聚合酶链反应和Western印迹评估MUC1的表达。使用标准铬 51 (51Cr) 释放测定和干扰素-g 释放测定来测量 MUC1 特异性 CTL 反应。结果：扩增的 MUC1 mRNA 可以有效转染树突状细胞。转染的 DC 在体外刺激 MUC1 特异性 CTL 反应方面非常有效。转染MUC1 mRNA的DC诱导的MUC1特异性CTL的功能受到主要组织相容性复合物（MHC）I类抗原呈递的限制。结论：转染MUC1 mRNA的DC刺激的CTL反应只能识别并裂解受MHC I类特异性抗原呈递限制的HLA-A2+/MUC1+ PC和其他靶细胞，为使用MUC1作为靶结构提供了临床前理论依据针对 PC 的免疫治疗策略。
Background and Aim: Mucin (MUC) 1 is an epithelial cell glycoprotein that is aberrantly overexpressed in many adenocarcinomas, including pancreatic cancer (PC), providing an ideal tumor-associated antigen and target for immunotherapy. In this study, we investigated whether the cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) transfected with amplified MUC1 mRNA could respond against PC in vitro.Methods: Amplified mRNA encoding MUC1 were transfected into DCs using electroporation with an optimized setting and the MUC1 expression were evaluated by quantitative real-time polymerase chain reaction and Western blot. The MUC1 specific CTL responses were measured using the standard chromium 51 (51Cr)-release assays and the interferon-g release assay.Results: Dendritic cells could be transfected with amplified MUC1 mRNA efficiently. The transfected DCs were remarkably effective in stimulating MUC1-specific CTL responses in vitro. The function of MUC1 specific CTLs, induced by MUC1 mRNA-transfected DCs, was restricted by major histocompatibility complex (MHC) class I antigen presentation.Conclusion: The CTL responses stimulated by DCs transfected with MUC1 mRNA could only recognize and lyse HLA-A2+/MUC1+ PC and other target cells under restriction by MHC class I-specific antigen presentation, providing a preclinical rationale for using MUC1 as a target structure for immunotherapeutic strategies against PC.