表达CD40配体的溶瘤痘苗病毒(CD40L-VV)抑制结直肠癌细胞生长并增强荷瘤小鼠T细胞抗肿瘤活性  

Oncolytic vaccinia virus expressing CD40L (CD40L-VV) inhibits colorectal cancer cell growth and enhances anti-tumor activity of T cells in tumor-bearing mice

作　　者：刘东涛[1] 马俊文[1] 丁波[1] 周海宁[1] LIU Dongtao;MA Junwen;DING Bo;ZHOU Haining(Department of Gastroenterological Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院胃肠外科,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2021年第7期602-607,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：宁夏自然科学基金(NZ16161);宁夏高等学校一流学科建设(宁夏医科大学国内一流建设学科临床医学)资助项目(NXYLXK2017A05)。

摘　　要：目的探索表达CD40配体的溶瘤痘苗病毒(CD40L-VV)对结直肠癌的抗肿瘤活性。方法通过将CD40L的基因序列整合至VV的DNA骨架中,获得CD40L-VV。通过体外感染肿瘤细胞,验证其溶瘤活性及检测CD40L的表达情况。利用CD40L-VV和单纯VV对结直肠癌移植瘤模型进行治疗后,监测肿瘤生长状况,并通过流式细胞术分析肿瘤浸润T细胞的表型,细胞因子分泌水平及增殖活性来反映CD40L-VV的抗肿瘤活性。结果 CD40L-VV可有效感染肿瘤细胞并表达CD40L,细胞存活实验表明VV及CD40L-VV对肿瘤细胞存在剂量依赖的裂解能力。体内移植瘤实验结果显示,CD40L-VV较VV抑制肿瘤生长的能力更强,与VV治疗组相比,CD40L-VV治疗组肿瘤浸润T细胞具有更强的细胞因子分泌能力,更强的增殖活性及更多的记忆细胞表型。结论工程化CD40L-VV可以显著抑制结直肠癌细胞生长,增强体内T细胞的抗肿瘤活性。Objective To explore the anti-tumor activity of oncolytic vaccinia virus expressing CD40L(CD40L-VV) against colorectal cancer. Methods The CD40L-VV was obtained by integrating the sequence of CD40L into the skeleton of oncolytic vaccinia virus(VV). The tumor cells were infected with VV and CD40L-VV to verify their oncolytic activity and the expression of CD40L in vitro. After the tumor model of colorectal cancer was treated with VV and CD40L-VV, the tumor growth was monitored, and the phenotype of tumor infiltrating T cells was analyzed by flow cytometry. The anti-tumor activity of recombinant oncolytic VV was also reflected by detecting the production of cytokine and the proliferation activity of tumor infiltrating T cells. Results Microscopic observation and Western blot assay showed that CD40L-VV could effectively infect tumor cells and express CD40L. Cell viability assay showed that VV and CD40L-VV had dose-dependent lytic ability against tumor cells. The results of tumor transplantation in vivo showed that CD40L-VV had stronger ability to inhibiting tumor growth than VV. Flow cytometry showed that tumor infiltrating T cells in the CD40L-VV group had stronger cytokine secretion ability, stronger proliferative activity and more memory cell phenotypes than those in the VV group. Conclusion CD40L-VV can significantly inhibit the growth of colorectal cancer cells and enhance the anti-tumor activity of T cells in vivo.

关 键 词：结直肠癌 溶瘤痘苗病毒(VV) 免疫疗法 CD40配体(CD40L) 

分 类 号：R735.37[医药卫生—肿瘤] R730.51[医药卫生—临床医学]