Human Papillomavirus: Detection of Viral DNA Sequences and Evidence for Molecular Heterogeneity in Metaplasias and Dysplasias of the Uterine Cervix

Abstract

Colposcopic-directed biopsies of the uterine cervix from 22 patients were analyzed for the presence of human papillomavirus (HPV) DNA and structural antigens. 11 of the biopsies were classified microscopically as mild dysplasia, 3 as moderate dysplasia, 1 as severe dysplasia, and 7 as squamous metaplasia. Nonstringent hybridization with a bovine papillomavirus type 1 DNA probe and immunocytochemical analysis with an antiserum against papillomavirus genus-specific structural antigens were performed on all specimens. Of the 11 mild dysplasias, both HPV DNA and structural antigens were detected in 5, only HPV antigens in 3, only HPV DNA in 1, and neither DNA nor structural antigens in 2. Both HPV DNA and structural antigens were present in the 3 cases of moderate dysplasia. Only HPV DNA sequences were detected in the single case of severe dysplasia. HPV DNA was detected in 2 cases of squamous metaplasia. The 5 remaining cervical biopsies showing squamous metaplasia, tissue from 3 placentas, and 6 cervical carcinomas were negative for HPV DNA and structural antigens. Restriction enzyme cleavage patterns of HPV DNA in the dysplasias suggested that there are multiple virus types or subtypes associated with cervical dysplasia. Stringent hybridization with a HPV type 11 (HPV-11) probe revealed that only 1 of 10 dysplasias contained sequences with homology to the probe. Of the remaining 9 dysplasias, 5 contained HPV sequences detected under nonstringent hybridization. 2 of 4 squamous metaplasias contained viral sequences which hybridized to the HPV-11 probe as well as 1 of 6 cervical carcinomas.

Cloning and characterization of the DNA of a new human papillomavirus from a woman with dysplasia of the uterine cervix

“…Cellular DNA suitable for restriction enzyme digestion and subsequent Southern blot hybridization studies ( Fig. 1) was prepared by proteinase K digestion and phenol extraction as described previously (21). The nucleic acid extract (1 to 10 ,ug) was digested with restriction endonuclease PstI or BamHI (Bethesda Research Laboratories, Inc.).…”

“…Hybridization and washes were modifications of a previously described procedure (21). The filters were baked under a vacuum for 30 min at 80'C and prehybridized in 4x SSC (lx SSC is 0.15 M NaCl plus 0.015 M sodium citrate) (34)-0.5 mM EDTA-10x Denhardt solution (8)-50 ,ug of tRNA per ml-20 mM sodium phosphate buffer (pH 7.4).…”

Detection of Papillomavirus DNA in Human Semen

“…cates that the NOD mouse is H-2K" and H-2Db (6). We also studied the reactivity of monoclonal antibodies to the I-A and I-E glycoprotein molecules and the interaction ofa conventional antiserum to Ia with NOD splenocytes (Table 1) (7)(8)(9)(10)(11)(12). As a control for the percentage of B lymphocytes in the spleen of each strain, we analyzed the reactivity of antimouse immunoglobulin.…”

Natural history of human papillomavirus infection of the anogenital tract

“…Although a number of agents, most notably herpes simplex virus 2, have been suspected of playing a role in cervical cancer, recent attention has focused on human papiUomavirus (HPV) as a possible etiologic agent in the genesis of CIN and invasive cancer. Immunologic and nucleic acid hybridization studies have demonstrated the presence of HPV structural antigens, DNA, or both in a high proportion of benign, premalignant, and malignant lesions of the cervix [2][3][4]. The discovery and molecular cloning of HPV types 16 and 18 from invasive cervical cancers [5,6], and the subsequent demonstration of the presence of these and closely related virus DNAs in the majority of CIN II-III lesions, invasive cervical cancers, cervical cancer lymph node metastases, and cervical cancer cell lines strongly suggests a causal relationship [5][6][7][8][9].…”