Expression in Escherichia coli of Seven DNA Fragments Comprising the Complete L1 and L2 Open Reading Frames of Human Papillomavirus Type 6b and Localization of the 'Common Antigen' Region

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Human papillomavirus types 6 and 11 (HPV-6 and HPV-11) are the major aetiological agents of condylomata acuminata. Serological studies of this disease have been difficult to perform and interpret because native, type-specific antigens have not been available. In particular, since these viruses have not been propagated in vitro and sufficient quantities of virions are not present in lesions, virus particles have been difficult to obtain. In the present study, we used HPV-11 particles, obtained from human tumours produced in athymic mice, as antigen in an ELISA to compare antibody responses between 46 patients with biopsyproven condylomata acuminata and 44 controls. The median [interquartile range] of the absorbance values for the condylomata acuminata and the control groups were respectively 0-324 [0-183, 1-029] and 0.118 [0-047, 0.286] (P= 0-0001). Thirty-three per cent of the absorbance values in the condylomata acuminata group were higher than any of those of the control group. Sera from patients whose biopsies contained the papillomavirus common antigen were more reactive than sera from patients whose biopsies did not contain it (P= 0-0014).This study demonstrates the presence of specific antibodies directed at native HPV-I 1 viral particles in the sera of patients with condylomata acuminata, and describes a test which can be used in future serological studies of this common sexually transmitted disease.

Section: W Bonnez and Othersmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Use of human papillomavirus type 11 virions in an ELISA to detect specific antibodies in humans with condylomata acuminata

Bonnez

Rin

Rose

et al. 1991

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“…Some of these sera are apparently type-specific; for example, antisera against an HPV-16 L1 fusion protein (Browne et al, 1988) does not recognize an HPV-6b L1 fusion protein, but antibodies against HPV-6b L1 (Tomita et al, 1987) cross-react with the major capsid protein of HPV-1 (Li et al, 1987). These cross-reactivities have been mapped to the mid-portion of the LI protein, as fusion proteins expressing HPV-6b L1 (amino acids 75 to 234 and 234 to 432) are recognized by antisera to BPV-1 and vice versa (Strike et al, 1989). However many of these sera have not been tested extensively against L1 proteins from different HPV types nor have they been tested for reactivity against HPV-infected tissue sections.…”

Section: Discussionmentioning

confidence: 99%

Identification of Immunogenic Regions of the Major Coat Protein of Human Papillomavirus Type 16 that Contain Type-restricted Epitopes

Cason¹,

Patel²,

Naylor³

et al. 1989

SUMMARYWe have identified regions of the major capsid protein, L1, of the human papillomavirus (HPV) type 16 , that are recognized by five monoclonal antibodies (MAbs) raised to a bacterial fusion protein containing residues 172 to 375 of HPV-16 L1. All five MAbs recognized HPV-16-infected tissue sections by immunohistochemistry, but not sections infected with HPV-la (cutaneous warts), HPV-6b or -11 (genital warts). MAbs 3D1, 5A4 and 1D6 also recognized HPV-2-infected sections (cutaneous warts); MAb 8C4 recognized only sections containing HPV-16. Four MAbs (8C4, 3D1, 1D6 and 5A4) recognized a synthetic peptide corresponding to residues 269 to 284 of HPV-16 L1; within this region a minimum antibody binding site was identified, a tripeptide 276 to 278. However the complete epitope appears to extend beyond these residues and beyond HPV-16 L1 (269 to 284). The fifth MAb, 1C6, recognized bacterial fusion proteins containing HPV-6b L1, -16 L1 or -18 L1 using immunoblots, yet appeared HPV-16-specific when tested on infected tissue sections. This MAb recognized five amino acids within a different region of HPV-16 L1 (residues 299 to 313). INTRODUCTIONPapillomaviruses infect avian, reptile and mammalian hosts, causing epithelial proliferation (Sundberg, 1987). Humans are susceptible to at least 58 types of papillomaviruses. Human papillomavirus (HPV) types 16, 18, 31, 33 and 52 which cause genital infections have been associated with high grade cervical intraepitheliat neoptasia lesions and invasive carcinoma, and other genital HPV types (e.g. HPV-6b and HPV-11) are more commonly associated with benign conditions such as genital warts (McCance, 1986). If the relationship between certain types of HPV and pre-invasive and invasive cancer of the cervix is proven to be of a causal rather than casual nature, rapid and accurate diagnosis of the type of HPV infection is likely to be important in the clinical management of patients with these diseases. At present, diagnosis of the exact type of HPV infection relies upon nucleic acid hybridization techniques, which are time-consuming and expensive when compared to immunohistochemical or serological methods of viral identification. A serological test for exposure to high risk HPV types would thus provide an easier method of identifying patients who may be at greatest risk of developing a malignant disease.The major capsid protein (L 1) of papillomaviruses, which is the most abundant virus-encoded protein, is highly conserved between different types, with amino acid homologies as high as 80~ existing between some L1 proteins from different papillomavirus types. As a consequence

“…In addition, intact virions are generally not present in large numbers in naturally infected tissues, and biophysical and antigenic characterization of HPV particles has been difficult to perform. Therefore, HPV proteins have been produced by recombinant DNA technology in prokaryotic expression systems Doorbar & Gallimore, 1987;Komly et al, 1986;Strike et al, 1989;Tomita et al, 1987) in order to study the molecular biology of HPV and evaluate host immune responses to infection.…”

mentioning

confidence: 99%

Expression of the full-length products of the human papillomavirus type 6b (HPV-6b) and HPV-11 L2 open reading frames by recombinant baculovirus, and antigenic comparisons with HPV-11 whole virus particles

Rose

Bonnez

Strike

et al. 1990

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The L2 open reading frames (ORFs) of human papillomavirus (HPV) types 6b and 11 were expressed as full-length non-fusion proteins in Spodoptera frugiperda (Sf-9) cells using recombinant baculovirus. Both proteins were detected on Western blots as immunoreactive bands which migrated with apparent Mrs of 76K and 78K, respectively, and contained both crossreactive and type-specific epitopes, as determined by polyclonal antisera directed against defined subregions of the HPV-6b and HPV-11 L20RFs. In addition, the minor capsid protein of HPV-11 particles co-migrates with the HPV-11 L20RF product and is immunoreactive with HPV-11 L2-specific antisera. These observations indicate that the anomalous electrophoretic mobilities of papillomavirus L20RF proteins can be explained without invoking post-transcriptional processing events and that the minor capsid protein of HPV-11 is antigenically and biophysically related to the HPV-11 L20RF product.