摘要
目的观察阳性药物D-盐酸青霉胺(D-pen)和链脲佐菌素(STZ)诱发的人免疫系统重建NPG(hu-NPG)小鼠的直接腘窝淋巴结试验(d-PLNA)反应,并比较其与NPG和BALB/c小鼠反应的异同,探索可提高药物超敏反应预测价值的模型动物。方法雌性NPG小鼠,1.8 Gy的X线辐照后,植入人脐血造血干细胞,制备hu-NPG小鼠,取外周血,进行淋巴细胞表型分析及淋巴细胞增殖实验。hu-NPG,NPG和BALB/c小鼠各10只,每种小鼠随机分成D-pen和STZ组,每组5只。各组分别在右后肢足趾部sc给予D-pen每只1 mg或STZ 0.75 mg,左后肢不做处理作为对照。注射后7 d处死小鼠,取左、右侧腘窝淋巴结称重,计算质量指数;将淋巴结制成单细胞悬液,计算细胞指数,并对部分淋巴结进行组织病理学检查,以比较d-PLNA反应。结果注射阳性药物D-pen和STZ后,各组小鼠一般状态良好,均未见全身毒性,且至处死时局部炎性刺激症状消失。D-pen和STZ均能诱导BALB/c小鼠出现典型的d-PLNA阳性反应,表现为注射侧淋巴结质量和细胞数量比对照侧增加,平均质量指数≥2或平均细胞指数≥5。而NPG小鼠和hu-NPG小鼠均未出现阳性反应。病理学检查发现,NPG和hu-NPG小鼠的腘窝淋巴结体积明显小于BALB/c小鼠,发育不完善。淋巴细胞表型分析发现,NPG小鼠仅有少量淋巴细胞,而hu-NPG小鼠外周血中人源性淋巴细胞以CD45^+CD19^+的B细胞为主,CD45^+CD3^+T细胞较少,且丝裂原不能刺激NPG和hu-NPG小鼠体内的人源或鼠源性淋巴细胞增殖。结论 D-pen和STZ所致的小鼠d-PLNA阳性反应依赖于体内正常的淋巴细胞数量及功能。NPG小鼠缺乏正常淋巴细胞,d-PLNA反应阴性。hu-NPG小鼠体内的人淋巴细胞功能不全,也不能诱发阳性反应。因此,hu-NPG小鼠尚不能成为d-PLNA的敏感动物。
OBJECTIVE To perform direct popliteal lymph node assay （d-PLNA） induced by D-penicillamine hydrochloride （D-pen） or streptozotocin （STZ）in humanized NPG （hu-NPG） mice with a reconstituted immune system, NPG mice and BALB/c mice respectively, and to compare the responses in these mice to elucidate the mechanism of d-PLNA. METHODS Female NPG mice were ir- radiated and transplanted with human hematopoietic stem cells isolated from cord blood to form hu-NPG mice. The number and function of peripheral blood lymphocytes in hu-NPG mice were detected by flow cytometry and lymphocyte proliferation test. The hu-NPG, NPG and BALB/c mice were randomly divided into D-pen group and STZ group （5 mice per group）, respectively. Then, the positive chemical D-pen （1 mg per mouse） or STZ （0.75 mg per mouse） was injected subcutaneously into the right hind footpad of these mice using the established d-PLNA protocol. On the day after injection, the mice were sacrificed. The bilateral popliteal lymph nodes （PLNs） were weighed. The difference of d-PLNA response was evaluated at 7 d after the injection of positive drugs. RESULTS All the NPG, hu-NPG and BALB/c mice behaved normally after the injection of positive drugs. No systemic toxicity was observed. The symptom of local irritation disappeared within 7 d. As in previous studies, the BALB/c mice showed a typical positive response of d-PLNA, as evidenced by the increase in both mass and cell count of the PLNs inihe treated lateral （mass index 〉2 and cellularity index 〉5）, while NPG mice and hu-NPG mice showed a negative resoponse. Pathological examinations demonstrated that the PLNs of NPG mice and hu-NPG mice had developmental defect in lymphoid tissue and were smaller in size than BALB/c mice. Phenotypic analysis of the peripheral blood lymphocytes in hu-NPG mice showed that the majority of human lymphocytes expressed B cell markers CD45＋CD19＋ （75.5±6.6）%, and only about （17.1±6.6）% of the lymphocytes expressed T cell markers CD45�
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2016年第8期839-847,共9页
Chinese Journal of Pharmacology and Toxicology
基金
山东省自然科学基金(ZR2012HM056)
山东省自主创新及成果转化专项(2014ZZCX02104)~~
关键词
药物超敏反应
直接腘窝淋巴结试验
人源化小鼠
drug hypersensitivity reaction
direct popliteal lymph node assay
humanized mice
