Wu J Y, Newton S, Judd A, Stocker B, Robinson W S
Department of Medicine, Stanford-University School of Medicine, CA 94305.
Proc Natl Acad Sci U S A. 1989 Jun;86(12):4726-30. doi: 10.1073/pnas.86.12.4726.
A nonvirulent Salmonella dublin flagellin-negative, aromatic-dependent live vaccine strain has been used to express hepatitis B virus surface antigen epitopes in an immunogenic form. The envelope proteins of the virion are encoded by the S gene, which contains the pre-S1, pre-S2, and S coding regions. Synthetic oligonucleotides corresponding to amino acid residues S-(122-137) and pre-S2-(120-145) were inserted in-frame into the hypervariable region of a cloned Salmonella flagellin gene, and the recombinant plasmids were introduced into a flagellin-negative aroA mutant live vaccine strain of S. dublin, SL5928. The flagellin gene was expressed in bacteria carrying the plasmids as detected by immunoblotting with anti-flagellin (H1-d) serum. Both the S and pre-S2 epitopes were detected in bacteria carrying the relevant plasmid by immunoblotting with anti-HBs (antibody to hepatitis B virus surface antigen) and anti-peptide antisera. Animals immunized intramuscularly or orally with the live recombinant bacteria developed antibodies specific to these hepatitis B virus epitopes as detected by ELISA.
一种无毒的都柏林沙门氏菌鞭毛蛋白阴性、芳香族氨基酸依赖的活疫苗菌株已被用于以免疫原性形式表达乙型肝炎病毒表面抗原表位。病毒粒子的包膜蛋白由S基因编码，该基因包含前S1、前S2和S编码区。将对应于氨基酸残基S-(122 - 137)和前S2-(120 - 145)的合成寡核苷酸读码框插入克隆的沙门氏菌鞭毛蛋白基因的高变区，然后将重组质粒导入都柏林沙门氏菌的鞭毛蛋白阴性aroA突变活疫苗菌株SL5928。通过用抗鞭毛蛋白(H1-d)血清进行免疫印迹检测发现，鞭毛蛋白基因在携带质粒的细菌中表达。通过用抗-HBs(抗乙型肝炎病毒表面抗原抗体)和抗肽抗血清进行免疫印迹检测，在携带相关质粒的细菌中检测到S和前S2表位。通过ELISA检测发现，用活重组细菌进行肌肉注射或口服免疫的动物产生了针对这些乙型肝炎病毒表位的特异性抗体。
