Characterization of murine polyclonal antisera and monoclonal antibodies generated against intact and denatured human papillomavirus type 1 virions

Abstract

Human papillomavirus type 1 (HPV1) virions, both as intact virion particles (IVP) and as detergentdenatured virions (DDV), were used to prepare polyclonal antisera and monoclonal antibodies (MAbs) in BALB/c mice. Anti-IVP antiserum contained type-specific HPV1 L2-reactive antibodies and no detectable HPV1 Ll-reactive antibodies. Anti-IVP MAbs recognized a linear epitope between L2 amino acids 102 and 108 (PIDVVDP). Anti-DDV antiserum contained type-specific HPV1 Ll-reactive and HPV1 L2-reactive antibodies. An anti-DDV MAb recognized a linear epitope between Li amino acids 127 and 133 (AENPTNY). HPVla Liand L2-encoded polypeptides expressed in Saccharomyces cerevisiae and by in vitro translation were equivalent in size to the major and minor virion capsid proteins, respectively.

Self-assembly of human papillomavirus type 1 capsids by expression of the L1 protein alone or by coexpression of the L1 and L2 capsid proteins

“…Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot were performed as described by Jenison et al (17,18). Murine monoclonal antibodies specific for lLl and 1L2 were obtained from David Baker at SUNY Stony Brook and have been characterized by Yaegashi et al (37). Immunofluorescent staining of HPV-1 Li and L2.…”

“…Like simian virus 40 and polyomavirus (2,3,14,31), the capsomeres of bovine papillomavirus type 1 and HPV-1 have been shown to possess fivefold symmetry (4) and therefore are likely to be pentamers of the major capsid protein. The Li open reading frame encodes the major capsid protein, and studies with virions and other systems expressing Li have identified a protein with an electrophoretic mobility consistent with its predicted size of approximately 55 kDa (8,12,25,30,33,37). A minor capsid antigen is encoded by the L2 ORF that also specifies a protein of 55 kDa.…”

“…A minor capsid antigen is encoded by the L2 ORF that also specifies a protein of 55 kDa. However, studies on isolated proteins from warts as well as L2 expression in a variety of systems, including in vitro transcription and translation, have shown that the L2 protein migrates with a molecular mass of approximately 72 kDa (8,12,20,34,37). The location of the minor capsid protein (L2 for HPV and VP2 and VP3 for simian virus 40 and polyomavirus) within virions has not been determined unambiguously.…”

Identification of type-specific and cross-reactive neutralizing conformational epitopes on the major capsid protein of human papillomavirus type 31

“…The inclusion of the corresponding VLPs in the composition of HPV vaccines has therefore to be considered. Both linear and conformational epitopes have been identified on the surface of HPV L1 VLPs [9,10,18,39,41,44]. It is now well established that conformational epitopes are responsible for neutralizing antibody production [7,14,21,42,43].…”

Human papillomavirus (HPV) type distribution and serological response to HPV type 6 virus-like particles in patients with genital warts

“…Many of the initial studies of serologic response to HPV employed recombinant DNA methods to clone and express late region 1 (L1) or 2 (L2) proteins with bacterial fusion proteins (16,23,49,50). Recently, the predominant humoral response to HPV virions has been shown to be directed against conformationally dependent structures that were not duplicated in bacterial fusion systems (12,17).…”