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

摘要

In order to study human acute B-lymphoblastic leukemia (B-ALL) in vivo, a novel xenotransplant model was established by using neonaial NOD/SCID/IL2 receptor γ chainmall 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/IL2γmall 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 multi parameter 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 humanCms+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.%本研究旨在应用NOD/SCID/Ⅱ2rγnull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/Ⅱ2rγnull新生期(出生48h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 - CD- 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( huCI45+ 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.