Abstract: Abstract: Objective To establish aＲAG2 /IL2ＲG double deficient mice model and the preliminary application．Method ＲAG2 deficient mice were crossed with IL2ＲG deficient mice． The genotype of hybrid mice was determined by PCＲ． The percentages of CD3 + ，CD19 + andCD49b + were detected y FCM in terms of its T，B lymphocyte function and NK cell percentage． The physiological and biochemical parameters of blood and major organs weight were observed． Ｒesult ＲAG2 and IL2ＲG deficient mouse were bred respectively，and ＲAG2 / IL2ＲG hybrid mice model was successfully established． Established stable PCＲ identification methods of ＲAG2 / IL2ＲG double genedeficient mice by PCＲ methods． The ＲAG2 /IL2ＲG hybrid mice lacks T cells，B cells，the percentage of NK cells was very low，and the difference was significant ( P ＜ 0. 05) ． Blood biochemical tests showed that compared with the same week-old C57BL /6， serum biochemical parameters of ＲAG2 /IL2ＲG double deficient mice was not significant changes，the difference was not significant ( P ＜ 0. 05 ) ． In the 18 blood physiological indices of ＲAG2 /IL2ＲG double deficient mice，Ｒed blood cell ( ＲBC) and hemoglobin ( HGB) content is low，and the difference was significant ( P ＜ 0. 05) ，White blood cell ( WBC) ，lymphocytes ( LYMPH) ， lymphocyte ratio ( LYM) ，monocytes，neutrophils count ( NEUT) index were significantly lower，and the difference was significant( P ＜ 0. 01) ． The main organ weight displayed the weights of spleen，thymus and mesenteric lymph node are lower than wild-type C57BL /6 mice，the difference was significant ( P ＜ 0. 05) ． The experiments of humanhepatoma cells ( tissue ) transplants show，ＲAG2 /IL2ＲG double gene-deficient mice tumor transplant success rate and growth rate were significantly higher，compared with the parental single gene-deficient mice and the difference was significant( P ＜ 0. 01) ． Conclusion The mice model of ＲAG2 /IL2ＲG double deficient and a stable system of identification are successfully established． The double deficient mouse provide a good animal model for patient-derived tumor xenograft model．

Key words: <p>ＲAG2, IL2ＲG, immunodeficient mouse, immune cells, tumorxenograft</p>

摘要： 摘要: 目的建立ＲAG2 /IL2ＲG 双基因缺陷的CＲG 小鼠杂交群体。方法将ＲAG2 基因缺陷小鼠与IL2ＲG 基因缺陷小鼠分别进行繁殖，选取ＲAG2 基因缺陷雄鼠ＲAG2( － / － ) 与IL2ＲG 基因缺陷雌鼠IL2ＲG( － / － ) 进行配对，培育杂交后代，通过PCＲ 扩增基因组ＲAG2 和IL2ＲG 基因进行鉴定; 应用流式细胞仪分析检测ＲAG2 /IL2ＲG 双基因缺陷小鼠外周血T 细胞( CD3 + ) 、B 细胞( CD19 + ) 和NK 细胞( CD49b + ) 含量; 并测定8—12 周龄ＲAG2 /IL2ＲG 双基因缺陷小鼠血液生理生化及主要脏器重量指标。结果成功繁育ＲAG2 基因缺陷小鼠和IL2ＲG 基因缺陷小鼠，筛选获得稳定的ＲAG2 /IL2ＲG 双基因缺陷小鼠并成功保种和扩群; 该双基因缺陷小鼠外周血淋巴细胞中T 细胞、B 细胞，NK 细胞比例显著降低( P ＜ 0. 05) ; 与相同周龄野生型C57BL/6 相比9 项血清生化指标无明显变化( P ＞ 0. 05) ; 18 项血液生理指标中红细胞( ＲBC) 和血红蛋白( HGB) 含量显著降低( P ＜ 0. 05) ; 白细胞( WBC) 、淋巴细胞数( LYMPH) 、淋巴细胞比率( LYM) 、单核细胞、中性细胞数( NEUT) 降低极显著( P ＜ 0. 01) ; 脾脏重量显著均低于野生型C57BL/6 小鼠( P ＜ 0. 05) ; 人肝肿瘤传代细胞移植于ＲAG2 /IL2ＲG 双基因缺陷小鼠后，肿瘤移植成功率和生长速率均明显高于亲本单基因缺陷小鼠( P ＜ 0. 05) ，成功获得了临床肝癌病人肿瘤组织的异种移植模型。结论 成功构建筛选出ＲAG2 /IL2ＲG 双基因缺陷小鼠杂交群体。

关键词: <, p>, ＲAG2, IL2ＲG, 免疫缺陷小鼠, 免疫细胞, 肿瘤移植<, /p>