人免疫重建NOD/SCID小鼠模型的建立

目的探讨NOD/SCID（nonobese diabetic/severe combined immunodeficien）t小鼠人免疫重建模型的建立方法和免疫特性.方法 16只NOD/SCID小鼠随机分成实验组和对照组,每组8只.Ficoll密度梯度离心法分离人外周血单个核细胞（peripheral blood mononuclear cell,PBMC）,通过腹腔注射移植给实验组小鼠,空白对照组小鼠每只腹腔注射无菌PBS,第4、8和12周时,流式细胞术检测小鼠外周血中人的CD3＋T、CD19＋B淋巴细胞,ELISA法测定小鼠血清中人IgG含量,免疫组织化学染色检测小鼠脾脏和肝脏中人CD3＋T、CD19＋B淋巴细胞浸润情况.结果移植PBMC 4周后,实验组NOD/SCID小鼠外周血中人CD3＋T、CD19＋B的细胞分别为85.6%、76.7%,并且在小鼠脾脏组织中也可以检测到人CD3 T、CD19 B淋巴细胞.移植4、8及12周后小鼠血清中人IgG含量分别达到（863±12.5）μg/mL、（1217±16.7）μg/mL、（958±13.1）μg/mL.结论用人外周血PBMC经腹腔注射可以成功建立人免疫重建NOD/SCID小鼠模型,方法简便可靠.

Establishment of Human Immune Reconstitute NOD/SCID Mice Model

Objective To study the establishment method of human immune reconstitute animal model in NOD/SCID mice and the characteristics of immunologic reconstitution.Methods 16 NOD/SCID mice were randomly divided into experimental group（n=8） and control group（n=8）.The peripheral blood mononuclear cells（PBMCs）were isolated from fresh peripheral blood of healthy people using Ficoll lymphocyte separating medium,and then were transplanted into experimental mice by intraperitoneal injection.Mice in control group was injected PBS buffer.In the 4th,8th and 12th weeks,human CD3 T and CD19 B lymphocytes in the blood of mice were detected by flow cytometry,human IgG protein content in the blood of mice was measured by ELISA method and the infiltration of human CD3 T and CD19 B lymphocytes in mice spleen and liver were detected by immunohistochemical staining.Results After 4 weeks,human CD3 T and CD19 B lymphocytes in peripheral blood of experimental mice were respectively 85.6% and 76.7% of the total monocytes,and were also detected in mice spleen.The amount of human IgG in serum of experimental mice after 4,8 and 12 weeks of transplantation were respectively（863±12.5） μg/mL,（1217±16.7） μg/mL and（958±13.1） μg/mL.Conclusions Human immune reconstitute NOD/SCID mice model can be successfully established by intraperitoneal injection of human PBMC.The method is simple and reliable.