Expression and immunogenicity of V3 loop epitopes of HIV-1, isolates SC and WMJ2, inserted in Salmonella flagellin.
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.
Cattozzo
E M
EM
Institute of Histology and General Embryology, School of Medicine, University of Ferrara, Italy.
Stocker
B A
BA
Radaelli
A
A
De Giuli Morghen
C
C
Tognon
M
M
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005408:Flagellin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D015483:HIV Antibodies / Q000097:blood*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D015497:HIV-1 / Q000737:chemistry / Q000276:immunology*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012475:Salmonella / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D005408:Flagellin