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郭文文,师长宏,李苗苗,乔天运,赵菊梅,张彩勤.前列腺癌免疫人源化小鼠模型的构建及鉴定[J].中国实验动物学报,2020,28(6):759~764.
前列腺癌免疫人源化小鼠模型的构建及鉴定
Establishment and identification of an immune-humanized mouse model of prostate cancer
投稿时间:2020-07-13  
DOI:10. 3969 / j.issn.1005-4847. 2020. 06. 004
中文关键词:  外周血单个核细胞  免疫人源化  前列腺癌  小鼠
英文关键词:peripheral blood mononuclear cells  humanization of immunity  prostate cancer  mouse
基金项目:
作者单位E-mail
郭文文 1. 延安大学医学院,陕西 延安 716000
2. 空军军医大学实验动物中心,西安 710032 
2399316897@ qq.com 
师长宏 空军军医大学实验动物中心,西安 710032  
李苗苗 空军军医大学实验动物中心,西安 710032  
乔天运 空军军医大学实验动物中心,西安 710032  
赵菊梅 延安大学医学院,陕西 延安 716000 jmz2003.stu@ 163.com 
张彩勤 空军军医大学实验动物中心,西安 710032 zhangcaiqin-beibei@ 163.com 
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中文摘要:
       目的 构建免疫系统人源化的小鼠模型,并移植入人前列腺癌细胞系,以期用于前列腺癌免疫治疗药物研究。 方法 利用 Ficoll 密度梯度离心法, 从新鲜的人外周血中分离出单个核细胞 ( peripheral blood mononucear cells,PBMC),经尾静脉注射 NPG 小鼠,构建具有人免疫系统的小鼠模型。注射后第 3、4、5、6 周分别采用断尾法采集小鼠外周血,并用流式细胞仪监测小鼠外周血中人 CD45 + CD3 + T 细胞的动态变化水平。在注射后 第 3 周给予小鼠皮下移植 1 × 107 个前列腺癌 22Rv1 细胞,每周两次监测肿瘤生长情况。当肿瘤生长至 1000 mmP>3P> 左右,对小鼠施行安乐死;利用流式细胞术、免疫组化、HE 染色等方法分析人源化小鼠外周血、脾及肿瘤组织中人免疫细胞的浸润情况。 结果 模型小鼠在移植人 PBMC 后 3 ~ 6 周期间,外周血中检测到较高水平的 CD45 + CD3 + T 细胞。第 6 周时处死小鼠,HE 染色和免疫组化染色结果表明,模型小鼠的脾和肿瘤组织中具有人 CD4 +、CD8 + T 细胞的浸润。 结论 成功构建了前列腺癌免疫人源化小鼠模型,其外周血、脾及前列腺癌肿瘤组织中均有高水平的人源性 T 细胞浸润,为下一步构建良好的前列腺癌免疫治疗临床前模型奠定了基础。
英文摘要:
       Objective An ideal animal model for the study of prostate cancer (Pca) immunotherapy is needed. We aimed to establish a mouse model humanized for the immune system and then transplant these mice with human Pca cells. Methods To humanize mice with a reconstructed human immune system, peripheral blood mononuclear cells (PBMCs) were separated from fresh human peripheral blood by density gradient centrifugation and injected into NOD-scid Il2rg- / - mice via the caudal vein. Peripheral blood from these mice was collected at the 3rd, 4th, 5th and 6th weeks following injection, and dynamic changes in human CD45+CD3+ T cells were monitored using flow cytometry analysis. At the 3rd week after humanization, cells from the Pca cell line 22Rv1 were subcutaneously implanted into the right flank of humanized mice. Tumor growth was monitored twice a week. When the tumor grew to about 1000 mmP>3P> , the mice were euthanized. The infiltration of human immune cells in the peripheral blood, spleen and tumor tissues of humanized mice was analyzed by flow cytometry, immunohistochemistry (IHC) and hematoxylin and eosin (HE) staining. Results High levels of CD45+CD3+ T cells were detected in the peripheral blood of mice from 3 to 6 weeks after PBMC transplantation. The mice were euthanized at the 6th week and the result of HE and IHC staining demonstrated that the spleen and tumor tissues of the mice were infiltrated by human immune cells CD4+ and CD8+ T cells. Conclusions The immune-humanized mouse model of Pca was successfully established. This model showed high levels of human immune T cells in the peripheral blood, spleen and Pca tissues, laying a foundation for the construction of an ideal preclinical model of immunotherapy for Pca.
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