白血病小鼠H-2全不相合微移植模型的建立及鉴定  

Establishment and Identification of a H-2 Completely Mismatched Microtransplantation Model of Leukemia Mouse

作　　者：吴惠惠[1] 刘铁强[1] 孙雪冬[1] 黄晓梅[1] 张锐[1] 刘志强[1] 满秋红[1] 黄雅静[1] 孙琪云[1] 左红莉[1] 乔建辉[1] 余长林[1] 胡锴勋[1] 艾辉胜[1] 郭梅[1] 

机构地区：[1]军事医学科学院附属医院血液科,北京100071

出　　处：《中国实验血液学杂志》2014年第3期779-784,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(861739);国家自然科学基金重点项目(81130054);首都临床特色应用研究(Z1211070001012082)

摘　　要：本研究旨在建立白血病小鼠H-2全不相合微移植模型并进行鉴定。受、供鼠分别为雌性BALB/c和雄性C57BL/6J,H-2为全不相合。受鼠微移植前5 d静脉接种WEHI-3细胞约1×106个,微移植前3 d开始给予MA化疗(米托蒽醌+阿糖胞苷),0 d末次化疗后8 h之内回输经G-CSF动员的供体脾单个核细胞,回输细胞数分别为(3、6、12)×107个。化疗对照组用等量生理盐水。所有受鼠不予GVHD预防。比较各组早期死亡率、白细胞恢复及白血病负荷情况。RT-PCR法检测微移植后供体嵌合率。从临床表现和病理情况综合评价微移植组小鼠GVHD情况。结果表明,化疗对照组早期死亡率为25%,(3、6、12)×107组分别为16.67%、8.33%、8.33%。(3、6)×107组白细胞恢复明显优于化疗对照和12×107组(P<0.05)。白血病负荷(3、6、12)×107组明显低于化疗对照组(P<0.01),而(6、12)×107组明显低于3×107组(P<0.05)。微移植后供体成分在2周之内以微嵌合形式存在。微移植组小鼠均未发生明显GVHD证据。结论:成功建立了白血病小鼠H-2全不相合微移植模型,为后续研究及临床微移植相关研究提供了基础。This study was purposed to establish and identify a H-2 completely mismatched microtransplantation model of leukemia mouse. The recipients were female BALB/c mice, while donors were male C57BL/6J mice. Recipients were inoculated intravenously with 1 x 106 of WEHI-3 cells, a cell line of myelomonocytic leukemia. Donors received 100 txg/kg G-CSF mobilization through hypodermic injection, every 12 hours, and it last 5 days. Chemotherapy regimens was MA （ mitoxantrone ＋ cytarabine）, and it last 4 days. Recipients were given chemotherapy conditioning without GVHD prophylaxis after inoculation of leukemic cells for 2 days, and within 8 hours after last chemotherapy received do- nor mobilized spleen mononuclear cells （sMNC）. The number of sMNC was （3,6,12） x 10^7, respectively. The early death rate, recovery level of WBC in peripheral blood and leukemia load were compared between chemotherapy and mi- crotransplantation groups. The donor chimerism was detected by RT-PCR. From the clinical manifestation and pathologi- cal features, the GVHD in recipients was evaluated. The results showed that the early mortality in chemothearpy group was 25% , meanwhile those in the （3,6,12） x 10^7groups were 16.67% ,8.33% ,8.33%, respectively. The（3,6） x 10^7 groups has a stronger hematopoietic recovery capability than that in chemotherapy and 12 x 10^7 groups（ P 〈 0.05 ）. There were more leukemic cells in chemothearpy mice than that in microtransplantation mice（P 〈0. 01 ）, and （ 12,6） x 10^7 groups had lower leukemia load than that in 3 x 10^7 group（P 〈 0.05）. No signs of GVHD were observed in microtrans- plantation mice. The donor microchimerism could be discovered at eraly 2 weeks after donor cell transfusion. It is concluded that a H-2 completely mismatched microtransplantation model of leukemia mouse has been successfully established, and it will provide a experimental base for studing microtransplantation in clinic.

关 键 词：白血病小鼠 H-2全不相合 微移植 微移植模型 微嵌合 

分 类 号：R733.7[医药卫生—肿瘤]