摘要
目的利用免疫缺陷小鼠构建胃癌患者来源的肿瘤组织异种移植(PDX)的免疫模型,分析原代及传代后小鼠模型肿瘤组织中T细胞的浸润情况。方法取胃癌患者的新鲜肿瘤组织样本,移植至免疫缺陷小鼠皮下,建立免疫PDX模型。收集移植前后及再次移植传代后的肿瘤组织,苏木精-伊红染色分析组织形态结构的改变,免疫组织化学分析CD3 T细胞的变化。流式细胞术分析荷瘤小鼠外周血中人CD8^(+)CD3^(+)T细胞比例的变化。结果3例胃癌组织中,1例成功移植。在皮下移植及再次传代过程中,肿瘤组织仍可维持形态学和组织学结构特征。免疫组织化学结果显示,肿瘤组织浸润的淋巴细胞数量在初次移植后逐渐减少,再次移植后完全丢失。初次移植后,荷瘤小鼠外周血中人源的CD3^(+)CD8^(+)T细胞比例呈现先上调后降低的变化趋势,持续存在时间可达60 d。结论成功建立人胃癌免疫PDX模型,初次移植的肿瘤组织及小鼠外周血的T细胞可存活60 d以上,提示该模型可用于进行免疫机制研究及免疫相关药物的筛选。
Objective To establish immune models with patient-derived gastric cancer xenograft(PDX)in NCG mice and analyze infiltrating T cells in tumor tissues derived from primary tumor and engrafts.Methods Fresh surgical specimens of gastric cancer were subcutaneously transplanted into NCG mice to establish the immune PDX model.Morphological structure of tumor tissues was analyzed by H&E staining and the infiltrating of human CD3 T cells were detected by immunohistochemistry in primary tumor tissues and engrafts.The percentages of CD8^(+)CD3^(+)T cells in the peripheral blood of NCG mice were determined by flow cytometry.Results One immune PDX model of gastric tumor was successfully established in NCG mice and obvious growth was observed 6 weeks post transplantation.After 12 weeks,engraft was isolated and transplanted into new NCG mice.No change of morphological structure and biology characteristics was found during first transplantation and subsequent passage.Tumor infiltrating lymphocytes(TIL)was gradually reduced after first transplantation and was almost disappeared after subsequent passage.In addition,the percentage of human CD3^(+)CD8^(+)T cells was increased in mice peripheral blood of immune PDX models after first transplantation,and then gradually decreased.T cells maintained in the blood up to 60 days after first transplantation.Conclusions Immune PDX models was successfully established in NCG mice.Human T cells maintained in primary engraftment and mouse peripheral blood for at least 2 months in gastric immune PDX model,indicating that this model could be used for immune mechanism research and immune-related drug screening.
作者
陈玲
刘英
朱艳阳
胡莉
张秋玉
CHEN Ling;LIU Ying;ZHU Yanyang;HU Li;ZHANG Qiuyu(Institute of Immunotherapy,Fujian Medical University,Fuzhou 350122,China;Department of Immunology,School of Basic Medical Sciences,Fujian Medical University,Fuzhou 350122,China)
出处
《福建医科大学学报》
2021年第4期287-292,共6页
Journal of Fujian Medical University
基金
福建省自然科学基金项目(2019J05074)
福建省中青年教师教育科研项目(JAT160210)。
