Identification of L2 open reading frame gene products of bovine papillomavirus type 1 using monoclonal antibodies.
Four hybridoma cell lines producing monoclonal antibodies (MAbs) to bovine papillomavirus type 1 (BPV-1) L2 open reading frame (ORF) gene products have been established from mice immunized with a BPV-1 L2-beta-galactosidase fusion protein. Hybridomas were selected and cloned (from over 700 hybridomas) on the basis of specific reactivity of supernatant fluids with BPV-1 L2 epitopes on disrupted BPV-1 particles and L2-beta-galactosidase fusion proteins by ELISA and Western blotting, and with acetone-fixed frozen sections of BPV-1-induced fibropapillomas by immunofluorescence. These MAbs were not reactive with intact BPV-1 particles or BPV-1 L1-beta-galactosidase fusion proteins by ELISA or with beta-galactosidase by ELISA and Western blotting. The four MAbs detected viral structural proteins of Mr 76K, 68K and possibly 55K in purified BPV-1 preparations by Western blotting. Two of the four MAbs were cross-reactive with BPV-2-induced fibropapillomas. These findings suggest that (i) the BPV-1 L2 ORF encodes the minor capsid protein(s), (ii) the gene products of the BPV-1 L2 ORF have Mr values of 76K, 68K and possibly 55K, (iii) minor capsid epitopes are internal to the BPV-1 particle, and (iv) MAbs reactive with genetically engineered truncated BPV-1 L2 ORF gene products can distinguish between BPV-1 and BPV-2 productive infections.
Jin
X W
XW
Department of Pathology, Georgetown University School of Medicine, Washington, D.C. 20007.
Cowsert
L M
LM
Pilacinski
W P
WP
Jenson
A B
AB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis; D015153:Blotting, Western; D052688:Bovine papillomavirus 1 / Q000235:genetics* / Q000276:immunology; D002213:Capsid / Q000235:genetics / Q000276:immunology; D003062:Codon / Q000235:genetics* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005260:Female; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D027383:Papillomaviridae / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014412:Tumor Virus Infections / Q000276:immunology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D003062:Codon; D000939:Epitopes; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins