Distribution and Specific Identification of Papillomavirus Major Capsid Protein Epitopes by Immunocytochemistry and Epitope Scanning of Synthetic Peptides

Abstract

Monoclonal (MAbs) and polyclonal antibodies were produced against the major capsid protein of detergent-disrupted, purified bovine papillomavirus type 1 (BPV-1). The precise locations of the corresponding epitopes were identified by the reactivity of MAbs and selected polyclonal antibodies with synthetic, overlapping, hexameric peptides corresponding with 95% of the BPV-1 major capsid protein. The topography of these epitopes was determined by reactivity of antibodies with intact (conformational and nonconformational surface epitopes) and disrupted (external or internal nonconformational epitopes) BPV-1 virions. The distribution of epitopes in various papillomaviruses of 13 different species was determined by reactivity of the MAbs and polyclonal sera with productively infected, formalin-fixed papillomas, fibropapillomas, and fibromas. Epitope scanning, using MAbs and polyclonal antisera, resulted in the precise location of BPV-1 hexameric epitopes that could be correlated with their topography on the capsid and distribution in papillomatous lesions of various species.

Feline Papillomas and Papillomaviruses

“…8,12,22 A panel of mouse monoclonal antibodies directed against linear epitopes of the bovine papillomavirus type-1 (BPV-1) L1 and L2 gene products that are phylogenetically conserved to various degrees was also used. 8,12,26 The reaction was developed by a modification of the avidinbiotin complex technique.…”

“…8,12,22 A panel of mouse monoclonal antibodies directed against linear epitopes of the bovine papillomavirus type-1 (BPV-1) L1 and L2 gene products that are phylogenetically conserved to various degrees was also used. 8,12,26 The reaction was developed by a modification of the avidinbiotin complex technique. 12 Positive controls included bovine cutaneous fibropapillomas containing either BPV-1 or BPV-2 and a canine oral papilloma containing the canine oral papillomavirus, all of which were typed by RFLP analysis from DNA in frozen biopsies.…”

“…Papillomavirus cross-reactive antigens, several of which are recognized by carefully characterized monoclonal antibodies, 8,12 were identified within the nuclei of koilocytotic cells (Figs. 1B, D, 3C, 4B).…”

“…Different PVs that infect a single species can be genotyped by various polymerase chain reaction methods, restriction fragment length polymorphisms (RFLPs), hybridization studies, and serotyping by patterns of antibody reactivities with linear and/or conformational capsid epitopes. 8,12 A case series of oral papillomatosis affecting the ventral tongues of Asian lions (Panthera leo persica) identified another feline papillomavirus that had a different epitope conservation pattern and genomic DNA restriction pattern from the domestic cat cutaneous papillomavirus. 23 This study compares the first two reported feline papillomaviruses 3,23 in relation to the pathology and conservation of capsid epitopes with six potentially new feline papillomaviruses.…”

Expression of human papillomavirus type 16 E6 protein by recombinant baculovirus and use for detection of anti-E6 antibodies in human sera

“…It has been established from studies on bovine papillomaviruses that antibodies generated against denatured capsid proteins are broadly cross-reactive with a wide range of papillomavirus types, whereas antibodies raised against intact virus particles tend to be type-specific (see Cowsert et al, 1987;Lim et al, 1990). Similarly, where bacterial fusion proteins or synthetic peptides have been used to generate antibodies against HPV coat proteins, broadly cross-reactive antisera are obtained (Strike et al, 1989;Christensen et al,, 1990).…”