耻垢分枝杆菌疫苗经灌胃免疫在小鼠体内的分布与安全性研究.docVIP

耻垢分枝杆菌疫苗经灌胃免疫在小鼠体内的分布与安全性研究 徐永柱1 吕琳2 1．重庆市卫生信息中心； 2.重庆医科大学附属第一医院消化内科 重庆 400016 【摘要】目的：探讨携带编码幽门螺杆菌UreB基因质粒(pLUB)的重组耻垢分枝杆菌经灌胃免疫在小鼠体内的分布和安全性。方法：将携带绿色荧光蛋白(GFP)基因质粒(pLUB)的重组耻垢分枝杆菌灌胃BALB／c小鼠后8周，处死小鼠，观察胃、心、肝、脾、肺和肾组织中荧光蛋白的分布，测定小鼠肌酐及ALT；处死小鼠，分离出肺、肾、肝并分别测定其重量及HE染色。结果：在BALB／c小鼠胃、脾组织中均检测到绿色荧光蛋白的表达，其他组织及对照组未检测到绿色荧光；小鼠体重、各脏器重量、各血清肌酐和ALT含量与对照组无差异(P0．05)。结论：携带GFP的重组耻垢分枝杆菌可在胃和脾组织分布，对小鼠无系统毒性作用。 【关键词】: 耻垢分枝杆菌疫苗; 安全性; 绿色荧光蛋白 The distribution and security of recombinant M.smegmatis in mice by gastric perfusion immunization Xu Yong Zhu1 ; Lü Lin2 (1.Chongqing Health Information Centre Chongqing 400014. 2. Department of Gastroenterology, The First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, ) 【Abstract】Objective： To observe the distribution and security of recombinant Mycobacterium smegmatis carrying the UreB gene of Helicobacter pylori by gastric perfusion immunization in mice．Methods：BALB／C mice were intragastricly immunized with recombinant M.smegmatis carrying the plasmid encoding green fluorescent protein(GFP) and UreB gene. The stomach, heart, liver, spleen, lungs and kidney was harvested 8 weeks after the immunization．The tissue slices were detected by Fluorescentmicroscope and HE stain. the body mass and organ mass were detected.The detected．Compared with the rM and control groups，the differencesf the body mass levels of ALT and Creatinine in serum were detected with Creatinine Kit and Alanine aminotransferase kit．Results：The distribution of GFP in gastric and spleen was and [2007]1号文 07-1-007） 作者简介：徐永柱（1965-），男，重庆江津人，副研究员，主要从事肿瘤的治疗相关研究，发表论文7篇，E-mail:XYZ850901@ Tel:023 通讯作者: 徐永柱 E-mail:XYZ850901@ organ mass were not significant.Conclusion：The recombinant M.smegmatis vaccine is safety to be used in mice and had no systemic toxicity. 【Key words】Mycobacterium smegmatis ；Green fluorescent protein；]。 1.1.2 实验动物雄性BALB／C小鼠，体重20±lg，购于重庆医科大学实验动物中心。 1.1.3 培养基Middle Brook 7H9 Broth （M7H9）,M7H9 Enrichment ADC均购于美国DIFCO公司；谷丙转氨酶试剂盒(赖氏法