人急性B淋巴细胞白血病-NOD/SCID/IL2rγnull新生小鼠异种移植模型的建立  

Establishment of Human Acute B-Lymphoblastic Leukemia NOD/SCID/IL2 Receptor γ Chain^(null) neonatal Mouse Xenotransplant Model

作　　者：孔圆[1,2] 刘艳荣[1] 刘开彦[1] 黄晓军[1] Fumihiko Ishikawa Mine Harada 

机构地区：[1]北京大学人民医院,北京大学血液病研究所,造血干细胞移植治疗血液病北京市重点实验室,北京100044 [2]日本九州大学医学研究院病态修复内科学,福冈812-8582 [3]日本RIKEN变态反应和免疫学研究中心人类疾病模式研究室,横滨230-0045

出　　处：《中国实验血液学杂志》2012年第3期762-768,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30800483);北京市科技计划项目(编号Z111107067311070)

摘　　要：本研究旨在应用NOD/SCID/IL2rγnull新生小鼠建立人急性B淋巴细胞白血病(B-ALL)的异种移植模型。NOD/SCID/IL2rγnull新生期(出生48 h之内)小鼠经亚致死剂量(100 cGy)137Cs全身照射后,由面静脉输注FACSAria流式细胞仪分选纯化的B-ALL患者骨髓CD3-CD4-CD8-CD34+CD19+细胞,于移植后8-12周用流式细胞术检测受鼠外周血、骨髓和脾脏中人源细胞的植入水平及其免疫表型,并通过HE染色评价人源B-ALL细胞在受鼠各组织器官中的迁移浸润能力。结果表明,在接受B-ALL患者CD34+CD19+细胞移植的受鼠外周血、骨髓和脾脏细胞中,不仅检测到了不同程度的人源B-ALL(huCD45+CD19+)细胞的植入〔(83.36±10.05)%,(93.88±5.05)%,(88.31±5.01)%〕,而且植入细胞具有与B-ALL患者相似的细胞形态和免疫表型特征。此外,人源B-ALL细胞广泛迁移浸润到受鼠的肝脏、肺脏、肾脏和脑等组织器官中。结论:NOD/SCID/IL2rγnull新生小鼠模型能够支持B-ALL患者CD34+CD19+细胞的高水平植入,是对人B-ALL进行体内功能性研究的新型异种移植模型。In order to study human acute B-lymphoblastic leukemia（B-ALL） in vivo,a novel xenotransplant model was established by using neonatal NOD/SCID/IL2 receptor γ chainnull mouse.The CD34＋CD19＋ bone marrow（BM） cells were sorted from the CD3-CD4-CD8-fraction of B-ALL patients by fluorescence-activated cell sorting（FACS）,and injected into 100 cGy irradiated neonatal NOD/SCID/IL2rγnull mice through facial vein.The engraftment and proliferation of human B-ALL cells were monitored by the presence of human CD45＋CD19＋ cells in peripheral blood（PB）.Human hematopoietic chimerism in PB,BM and spleen of the recipients was examined by multiparameter flow cytometry.Morphological analyses of FACS-sorted murine marrow cells were performed by using May-Grunwald-Giemsa staining.Furthermore,leukemia cell infiltration in the organs was evaluated by hematoxylin-eosin staining.The results indicated that the sorted CD34＋CD19＋ cells were able to initiate human B-ALL in vivo.The percentages of human CD45＋CD19＋ cells in PB,BM and spleen of the recipient mice were（83.36±10.05）%,（93.88±5.05）% and（88.31±5.01）%,respectively.Furthermore,the phenotype and morphology of the engrafted human cells were resemble to the original B-ALL cells from the patients.Similar to the clinical features,transplanted leukemic cells infiltrated into the organs,such as liver,lung,kidney and brain in the recipients.It is concluded that neonatal NOD/SCID/IL2rγnull mice can support efficient engraftment of the sorted CD34＋CD19＋ cells from human B-ALL for a long-term period.Human-mouse xenotransplant model using neonatal NOD/SCID/IL2rγnull mouse may provide an important system to study the biology of human B-ALL in vivo.

关 键 词：NOD/SCID/IL2rγnull小鼠 新生小鼠 异种移植 B-ALL 

分 类 号：R733.71[医药卫生—肿瘤] R457.7[医药卫生—临床医学]