A source of glycosylated human T-cell lymphotropic virus type 1 envelope protein: expression of gp46 by the vaccinia virus/T7 polymerase system.
Heterologous expression of the human T-cell lymphotropic virus type 1 (HTLV-1) envelope surface glycoprotein (gp46) in a vaccinia virus/T7 polymerase system resulted in the production of authentic recombinant gp46. Five differentially glycosylated forms of the surface envelope protein were produced by this mammalian system, as demonstrated by tunicamycin inhibition of N-glycosylation and N-glycan removal with endoglycosidase H and glycopeptidase F. These studies revealed that all four potential N-glycosylation sites in gp46 were used for oligosaccharide modification and that the oligosaccharides were mannose-rich and/or hybrid in composition. Conformational integrity of the recombinant HTLV-1 envelope protein was determined by the ability to bind to various HTLV-1-infected human sera and a panel of conformational-dependent human monoclonal antibodies under nondenaturing conditions. Furthermore, this recombinant gp46 was recognized by a series of HTLV-2-infected human sera and sera from a Pan paniscus chimpanzee infected with the distantly related simian T-cell lymphotropic virus STLVpan-p. Maintenance of highly conserved conformational epitopes in the recombinant HTLV-1 envelope protein structure suggests that it may serve as a useful diagnostic reagent and an effective vaccine candidate.
Gene Therapy and Molecular Virology Group, John P. Robarts Research Institute, London, Ontario, Canada.
J
J
Arp
M
M
LeVatte
J
J
Rowe
S
S
Perkins
E
E
King
C
C
Leystra-Lantz
S K
SK
Foung
G A
GA
Dekaban
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017123:Bacteriophage T7 / Q000235:genetics; D001666:Binding Sites, Antibody; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D000939:Epitopes / Q000276:immunology*; D015870:Gene Expression; D015686:Gene Products, env / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors; D006031:Glycosylation; D015481:HTLV-I Antibodies / Q000276:immunology; D015486:HTLV-I Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015490:HTLV-I Infections / Q000097:blood / Q000276:immunology; D006367:HeLa Cells; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000276:immunology*; D015367:Human T-lymphotropic virus 2 / Q000276:immunology; D006801:Humans; D007739:L Cells; D051379:Mice; D009844:Oligosaccharides / Q000276:immunology; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015682:Retroviridae Proteins, Oncogenic / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015303:Simian T-lymphotropic virus 1 / Q000276:immunology; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015686:Gene Products, env; D015481:HTLV-I Antibodies; D015486:HTLV-I Antigens; D009844:Oligosaccharides; D011993:Recombinant Fusion Proteins; D015682:Retroviridae Proteins, Oncogenic; D014764:Viral Proteins; C055266:gp46 protein, Human T-cell leukemia virus type I; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases