人过表达肝细胞生长因子间充质干细胞对小鼠的免疫原性和免疫毒性评价  

An Evaluation of Immunogenicity and Immunotoxicity of Human Overexpressing Hepatocyte Growth Factor Mesenchymal Stem Cells in Mice

作　　者：林庚 孙合强 王润生 陶梦雨 张乐统 徐国纲[2,4] LIN Geng;SUN Heqiang;WANG Runsheng;TAO Mengyu;ZHANG Letong;XU Guogang(Chinese PLA Medical School,Beijing 100853,China;Health Management Institute,The Second Medical Center,Chinese PLA General Hospital,Beijing 100853,China;Department of Laboratory Medicine,The Second Medical Center,Chinese PLA General Hospital,Beijing 100853,China;National Clinical Research Center for Geriatric Diseases,Beijing 100853,China;Pulmonary and Critical Care Medicine,The Second Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院,北京100853 [2]解放军总医院第二医学中心健康医学科,北京100853 [3]解放军总医院第二医学中心检验科,北京100853 [4]国家老年疾病临床医学研究中心,北京100853 [5]解放军总医院第二医学中心呼吸与危重症医学科,北京100853

出　　处：《标记免疫分析与临床》2024年第10期1927-1933,1940,共8页Labeled Immunoassays and Clinical Medicine

基　　金：北京市自然科学资金(编号:7242028)。

摘　　要：目的评价人过表达肝细胞生长因子间充质干细胞(hepatocyte growth factor-overexpressing dental pulp-stem cells,HGF-DPSCs)对小鼠的免疫原性和免疫毒性。方法采用BALB/c小鼠,雌雄各半,随机分组,每组6只,共6组,合计动物36只。设立第3天对照组(D3 PBS组)、第3天尾静脉注射DPSCs组(D3 DPSCs组)、第3天尾静脉注射HGF-DPSCs组(D3 HGF-DPSCs组)、第7天对照组(D7 PBS组)、第7天尾静脉注射DPSCs组(D7 DPSCs组)、第7天尾静脉注射HGF-DPSCs组(D7 HGF-DPSCs组)。全自动动物血液细胞分析仪检测外周血淋巴细胞、粒细胞、单核细胞,ELISA检测HGF、IgA、IgE、IgG、IgM、IFN-γ、IL-4、TNF-α表达水平,HE染色评价肝、肾、脾、肺组织的病理变化。结果D3 HGF-DPSCs组和D7 HGF-DPSCs组的血清人源HGF水平分别高于D3 DPSCs组和D7 DPSCs组,D3 PBS组和D7 PBS组不表达人源HGF。D7 PBS组较D3 PBS组、D7 DPSCs组较D3 DPSCs组、D7 HGF-DPSCs组较D3 HGF-DPSCs组,在淋巴细胞、炎症因子和器官毒性上均未观察到明显区别。各组间淋巴细胞总数、淋巴细胞百分比、总IgA、总IgE、总IgG、总IgM、IL-4、TNF-α、IFN-γ等指标差异均无统计学意义。同时HE染色结果表明经尾静脉注射HGF-DPSCs后,小鼠各脏器未出现明显病理损伤。结论腺病毒载体构建的HGF-DPSCs可在小鼠体内高表达HGF,同时HGF-DPSCs并无明显的免疫原性和免疫毒性,为HGF-DPSCs的后续研究和临床应用提供了直接的数据支撑和理论依据。Objective To evaluate the immunogenicity and immunotoxicity of human overexpressing hepatocyte growth factor-overexpressing dental pulp-stem cells(HGF-DPSCs)in mice.Methods A total of 36 BALB/c mice,comprising three males and three females per group,were randomly allocated to six experimental groups,with a control group on day 3(D3 PBS group),a group injected with DPSCs in the tail vein on day 3(D3 DPSCs group),a group injected with HGF-DPSCs in the tail vein on day 3(D3 HGF-DPSCs group),a control group on day 7(D7 PBS group),a group injected with DPSCs in the tail vein on day 7(D7 DPSCs group),and a group injected with HGF-DPSCs in the tail vein on day 7(D7 HGF-DPSCs group).A fully automatic animal blood cell analyzer was employed to detect peripheral blood lymphocytes,granulocytes and monocytes.ELISA method was applied to detect the expression levels of HGF,IgA,IgE,IgG,IgM,IFN-γ,IL-4 and TNF-α.HE staining was performed to evaluate the pathological changes in the tissues of the liver,kidney,spleen and lung.Results Serum concentrations of human HGF were found to be higher in the D3 HGF-DPSCs and D7 HGF-DPSCs groups than in the D3 DPSCs and D7 DPSCs groups,respectively.Furthermore,human HGF was not expressed in the D3 PBS and D7 PBS groups.There were no statistically significant differences in the total number of lymphocytes,percentage of lymphocytes,total IgA,total IgE,total IgG,total IgM,IL-4,TNF-α,and IFN-γamong these groups.In addition,histological examination results revealed that following the administration of HGF-DPSCs via the tail vein,no discernible pathological damage was observed in any of the studied organs.Conclusion The HGF-DPSCs can be constructed using adenoviral vectors and demonstrates the capacity to express HGF at a high level in mice model.Moreover,the HGF-DPSCs demonstrates no immunogenicity or immunotoxicity,providing strong data-driven supports and theoretical basis for subsequent research and clinical application of HGF-DPSCs.

关 键 词：肝细胞生长因子 间充质干细胞 免疫原性 免疫毒性 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]