摘要
目的使用NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju(NCG)小鼠建立人源化狼疮肾炎(lupus nephritis,LN)模型,检测小鼠免疫重建情况及自身抗体水平,并研究小鼠存活时间、肾脏病理表现与患者临床表现的关联。方法通过NCG小鼠尾静脉回输健康人或系统性红斑狼疮(systemic lupus erythematosus,SLE)合并肾损伤患者的外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs)建立HC、LN模型。用流式细胞术检测供体PBMCs及小鼠脾脏的人源CD45^+(h CD45^+)细胞、h CD3^+T细胞、h CD19^+B细胞、h CD4^+CD25highh CD127lowTreg细胞、h CD4^+h IFN-γ^+Th1细胞及h CD4^+h IL-17A^+Th17细胞比例,间接免疫荧光法测定抗核抗体(antinuclear antibody,ANA)水平,过碘酸雪夫(periodic acid-schiff,PAS)染色法观察小鼠肾脏病变。结果通过尾静脉回输移植人PBMCs,可以成功在NCG小鼠中实现人源化免疫重建,重建的人源化白细胞以T细胞为主,B细胞比例极少,但SLE患者PBMCs移植则可导致相对较高程度的B细胞重建;尽管供体回输前细胞具有一定比例的Treg细胞,但回输后并非所有小鼠脾脏均可检测到明显的Treg细胞重建(3/4 HC模型组小鼠Treg细胞重建明显,1/6 LN模型组小鼠Treg细胞重建明显);而健康人Th1细胞与Th17细胞在小鼠中重建不明显(2/2); LN模型组小鼠存活时间明显低于HC模型组小鼠,且根据PBMCs供体患者的临床病情严重程度,具有不同程度的肾炎表现。结论通过SLE患者PBMCs回输的方法在NCG小鼠中建立LN模型具有可行性,该模型为研究SLE合并LN患者的发病机制及个体化治疗提供了可能。
Objective The NCG mice were used to develop a humanized Lupus Nephritis（ LN） model,to investigate the immune reconstruction and the plasma autoantibodies in mice,and to observe the correlation between survival time,renal pathology and related patient suffered situation. Methods A LN model was set up by transfusing the peripheral blood mononuclear cells（ PBMCs） of healthy individuals or systemic lupus erythematosus（ SLE） patients combined with kidney injury into NCG mice. The proportion of human CD45^＋（ h CD45^＋） cells,h CD3^＋T cells,h CD19^＋B cells,h CD4^＋CD25^ high h CD127^low Treg cells,h CD4^＋h IFN-γ^＋Th1 and h CD4^＋IL-17 A^＋Th17 cells in donor PBMCs and mouse spleen were detected by flow cytometry. The level of anti-nuclear antibody（ ANA） was detected by indirect immunofluorescence and ELISA. The anti-double stranded DNA antibody（ anti-dsDNA） level was measured,and Periodic Acid-Schiff（ PAS） staining was used to observe the renal lesion in mice. Results Human PBMCs can be successfully transplanted into NCG mice to achieve humanized immune reconstitution,the rebuilting cells were mainly T cells,and the spleen of SLE patient had a higher degree of B cell reconstruction. Although the donor cells had a certain proportion of Treg cells before transplantation,Treg cell reconstruction cannot be detected significantly in all the spleen of mice（ 3/4 in HC model mice and 1/6 LN in model mice）,while Th1 and Th17 cells in healthy individuals were not rebuilt significantly in mice（ 2/2）. The survival time of SLE mice was shorter than that of HC model,and showed different degrees of renal injury. Conclusion It is feasible to develop a LN model in NCG mice by means of PBMCs backtransfusion originated from SLE patients,which could provide a possibility to study the pathogenesis and individualized treatment of SLE patients sufferd from LN.
作者
周昊天
孙晶晶
李雪
赵晓珍
张霞
何菁
孙晓麟
ZHOU Hao-tian;SUN Jing-jing;LI Xue;ZHAO Xiao-zhen;ZHANG Xia;HE Jing;SUN Xiao-lin(Department of Rheumatology and Immunology,Peking University People＇s Hospital,Beijing 101300,China)
出处
《微生物学免疫学进展》
2018年第5期36-43,共8页
Progress In Microbiology and Immunology
基金
国家自然科学基金(81471601)
国家自然科学基金(81601417)
关键词
人源化疾病模型
系统性红斑狼疮
狼疮肾炎
Humanized disease model
Systemic lupus erythematosus （SLE）
Lupus nephritis （LN）
