Zhang Hui, Hu Maozhi, Gu Zhiqiang, Jiao Xin'an, Geng Shizhong, Zhang Xiaoming, Pan Zhiming
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
Wei Sheng Wu Xue Bao. 2008 Jan;48(1):80-4.
To analyze the localization of attenuated Salmonella typhimurium after oral immunization. Prokaryotic expression plasmid pYA33-DsRed, carrying the RFP gene, was constructed and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria were named as X4550 (33-DsRed). The macrophage cell line RAW264.7 and bone marrow dendritic cell (BMDC) were invaded by X4550 (33-DsRed) in vitro. Furthermore, BALB/c mice were immunized with recombinant bacteria orally. RFP positive cells (RFP+ cells) were detected by Flow Cytometry (FCM) from spleen, liver, Mesenteric lymp node (MLN), Peyer's patch (PP), Inguinal lymph node (ILN). The invasion rate increased when the multiplicity of infection(MOI) were improved in this two kinds of cells respectively. After oral immunization with X4550 (33-DsRed), RFP+ cells were detected by FCM on 1d, 2d, 3d, 5d, 7d in spleen, liver, MLN, PP, ILN cells. The first day, RFP+ cells were detected in MLN and PP, and in PP at a higher rate of 1.4% than that of MLN. 0.4% RFP+ cells were detected the next day in ILN. On 3th day, the rates of RFP+ cells were increased in all of above tissues or organs and decreased on the 5th day. At the 7th day, RFP+ cells couldn't be detected in all tissues or organs tested. It is suggested that the invasion ability and the transfer through mucosal pathway and targeting to recognize immune tissue or organs are favor of the research in mucosal vaccine and the vaccine efficiency.
分析口服免疫后减毒鼠伤寒沙门氏菌的定位。构建携带红色荧光蛋白（RFP）基因的原核表达质粒pYA33-DsRed，并将其电转化至鼠伤寒沙门氏菌减毒株X4550中，重组菌命名为X4550（33-DsRed）。体外将X4550（33-DsRed）侵袭巨噬细胞系RAW264.7和骨髓树突状细胞（BMDC）。此外，用重组菌对BALB/c小鼠进行口服免疫。通过流式细胞术（FCM）检测脾脏、肝脏、肠系膜淋巴结（MLN）、派氏结（PP）、腹股沟淋巴结（ILN）中的RFP阳性细胞（RFP+细胞）。分别提高这两种细胞的感染复数（MOI）时，侵袭率增加。用X4550（33-DsRed）口服免疫后，在第1天、2天、3天、5天和7天通过FCM检测脾脏、肝脏、MLN、PP、ILN细胞中的RFP+细胞。第1天，在MLN和PP中检测到RFP+细胞，且在PP中的检出率更高，为1.4%，高于MLN中的0.4%。第2天在ILN中检测到0.4%的RFP+细胞。第3天，上述所有组织或器官中RFP+细胞的比率均升高，第5天下降。第7天，在所检测的所有组织或器官中均未检测到RFP+细胞。提示其侵袭能力、通过黏膜途径转移并靶向识别免疫组织或器官的特性有利于黏膜疫苗及疫苗效果的研究。
