Cattozzo E M, Stocker B A, Radaelli A, De Giuli Morghen C, Tognon M
Institute of Histology and General Embryology, School of Medicine, University of Ferrara, Italy.
J Biotechnol. 1997 Aug 28;56(3):191-203. doi: 10.1016/s0168-1656(97)00117-x.
Synthetic oligonucleotides corresponding to specific V3 loop portions of two HIV-1 isolates, SC and WMJ2, were expressed in the flagella of a Salmonella live-vaccine strain. Expression of the inserted epitopes in flagellin and their exposure at the surface of flagellar filaments were shown by immunoblotting and immunogold labeling with anti-flagellin (Salmonella d) and anti-HIV-1(IIIB) V3 loop peptide sera. Live recombinant Salmonella strains expressing either one of the two V3 loop inserts were administered intraperitoneally to BALB/c mice. All these animals developed antibodies specific for the heterologous glycoprotein 120 (gp120) of HIV-1 MN strain, as detected by enzyme-linked immunosorbent assays (ELISA), two of the sera had neutralizing activity against the heterologous HIV-1 MN strain. Moreover, oral administration of the live Salmonella recombinant strains to mice evoked specific IgA directed against gp120.
与两种HIV-1分离株SC和WMJ2的特定V3环部分相对应的合成寡核苷酸,在鼠伤寒沙门氏菌活疫苗株的鞭毛中表达。通过用抗鞭毛蛋白(鼠伤寒沙门氏菌d)和抗HIV-1(IIIB)V3环肽血清进行免疫印迹和免疫金标记,显示了插入表位在鞭毛蛋白中的表达及其在鞭毛丝表面的暴露。将表达两种V3环插入物之一的活重组沙门氏菌菌株腹腔注射给BALB/c小鼠。通过酶联免疫吸附测定(ELISA)检测,所有这些动物都产生了针对HIV-1 MN株异源糖蛋白120(gp120)的特异性抗体,其中两份血清对异源HIV-1 MN株具有中和活性。此外,给小鼠口服活沙门氏菌重组菌株可诱发针对gp120的特异性IgA。