Expression and immunogenicity in rats of recombinant adenovirus 5 DNA plasmids and vaccinia virus containing the HTLV-I env gene.
The complete human T-cell leukemia virus type I (HTLV-I) env gene was inserted into an expression cassette containing the adenovirus 5 major late promoter (Ad5-MLP). Recombinant Ad5-HTLV-I-env was obtained by homologous recombination in 293 cells simultaneously transfected by the expression cassette and the genomic DNA of Ad5. In vitro expression of the HTLV-I-env gene in the recombinant vector was detected by immunofluorescence and Western blotting. Functional expression of HTLV-I-env was confirmed by syncitium formation specifically in HeLa cells infected with Ad5-HTLV-I-env. Two immunization regimens against HTLV-I were tested in WKY and Fischer F-344 rats. The first involved WKY rats primed with Ad5-HTLV-I-env or naked DNA plasmids containing the HTLV-I-env gene and boosted with Ad5 containing the HTLV-I-env gp46 gene or with baculovirus-derived recombinant gp46. No antibody against HTLV-I was detected, while HTLV-I-specific cytotoxic T lymphocytes were recovered from all immunized groups but not from controls. The second approach involved Fischer F-344 rats primed and boosted with recombinant vaccinia virus containing the HTLV-I-env gene. Such rats developed antibodies against the HTLV-I env gp21 and gp46 (non-neutralizing). After challenge with human HTLV-I-producing cells (MT-2), both immunization regimens were found to induce partial protection.
Kazanji
M
M
Unité d'Epidémiologie des Virus Oncogènes, Institut Pasteur, Paris, France.
Bomford
R
R
Bessereau
J L
JL
Schulz
T
T
de Thé
G
G
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D003602:Cytotoxicity, Immunologic; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000276:immunology; D015752:Genes, env / Q000235:genetics / Q000276:immunology; D015481:HTLV-I Antibodies / Q000032:analysis; D006367:HeLa Cells / Q000821:virology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D011921:Rats, Inbred WKY; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000276:immunology
D017931:DNA Primers; D004279:DNA, Viral; D015481:HTLV-I Antibodies; D014759:Viral Envelope Proteins