Khan C M, Villarreal-Ramos B, Pierce R J, Riveau G, Demarco de Hormaeche R, McNeill H, Ali T, Fairweather N, Chatfield S, Capron A
Department of Pathology, University of Cambridge, United Kingdom.
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11261-5. doi: 10.1073/pnas.91.23.11261.
A vector has been constructed to allow genetic fusions of guest antigens via a hinge domain to the C terminus of the highly immunogenic C fragment of tetanus toxin. A fusion has been constructed with the gene encoding the protective 28-kDa glutathione S-transferase (EC 2.5.1.18) from Schistosoma mansoni. The recombinant vector has been electroporated into the nonvirulent Salmonella typhimurium aroA live vaccine strain SL3261. The corresponding chimeric protein is stably expressed in a soluble form in Salmonella as evaluated by Western blotting with fragment C and glutathione S-transferase antisera. Mice immunized intravenously with a single dose of the live recombinant bacteria elicit antibodies to both fragment C and glutathione S-transferase as detected by enzyme-linked immunosorbent assays. Furthermore, all of the mice were solidly protected when challenged with lethal doses of either tetanus toxin or the virulent Salmonella typhimurium strain C5. Mice have also elicited antibodies to fragment C and glutathione S-transferase after oral immunization. It may be that a live trivalent vaccine against typhoid, tetanus, and schistosomiasis is feasible.
已构建一种载体,用于通过铰链结构域将客体抗原与破伤风毒素高免疫原性C片段的C末端进行基因融合。已构建一种与编码曼氏血吸虫保护性28 kDa谷胱甘肽S-转移酶(EC 2.5.1.18)的基因的融合体。将重组载体电穿孔导入无毒的鼠伤寒沙门氏菌aroA活疫苗株SL3261。通过用C片段和谷胱甘肽S-转移酶抗血清进行蛋白质印迹分析评估,相应的嵌合蛋白在沙门氏菌中以可溶形式稳定表达。用单剂量的活重组细菌静脉内免疫小鼠,通过酶联免疫吸附测定法检测到小鼠产生了针对C片段和谷胱甘肽S-转移酶的抗体。此外,当用致死剂量的破伤风毒素或有毒的鼠伤寒沙门氏菌菌株C5攻击时,所有小鼠都得到了可靠的保护。口服免疫后,小鼠也产生了针对C片段和谷胱甘肽S-转移酶的抗体。或许一种针对伤寒、破伤风和血吸虫病的活三价疫苗是可行的。