Odile Croissant

High Prevalence of Papillomavirus-Associated Penile Intraepithelial Neoplasia in Sexual Partners of Women with Cervical Intraepithelial Neoplasia

To determine whether neoplastic cervical lesions in women are associated with papillomavirus infections in their sexual partners, we used a colposcope to examine male sexual partners of women with cervical flat condyloma (294 cases) or cervical intraepithelial neoplasia (186 cases), before and after 5 percent acetic acid was applied to the penis and the anogenital area. Condylomata acuminata, papules, and macules were observed in 309 of the 480 men (64.4 percent). In 204 of them (42.5 percent), macules or slightly elevated papules were detected only after application of acetic acid. Condylomata acuminata or lesions showing histologic features of condyloma were found in 121 partners (41.2 percent) of women with condyloma, but in only 10 partners (5.4 percent) of women with cervical intraepithelial neoplasia. Penile lesions showing histologic features of intraepithelial neoplasia were found in 61 partners (32.8 percent) of women with cervical intraepithelial neoplasia, but in only 4 partners (1.4 percent) of women with flat condyloma. Thirty-six (60 percent) of the 60 macules or papules tested contained papillomavirus DNA sequences. Human papillomavirus types 16 and 33 were almost exclusively found in penile intraepithelial neoplasia. Type 6, type 11, and the recently recognized type 42 were found in lesions showing features of condyloma or minimal histologic changes. As yet uncharacterized papillomaviruses were found in 15 percent of the specimens. These data support the concept that cervical carcinomas and precancerous lesions in women may be associated with genital papillomavirus infection in their male sexual partners.

Cutaneous warts clinical, histologic, and virologic correlations☆

Abstract Human-papillomavirus (HPV)-induced cutaneous lesions differ considerably in their clinical morphology. This was thought to depend mainly on anatomic location and other factors related to the host and to the age of the lesions. It was long-assumed that a single virus was responsible for all of these lesions. 1 Disclosure of the plurality of HPV types associated with skin warts 2–9 raised the problem of the relation of clinical morphology to the distinct types of HPV. This was first suggested by the characterization of distinct HPVs from different types of lesions: HPV1 from deep plantar warts, 3 HPV2 from common hand warts, 3 and HPV3 from plane warts and flat wart-like lesions of patients with epidermodysplasia verruciformis (EV). 5 At present, 39 types of HPVs are recognized, with several subtypes for many of them. 10 Some HPV types are specifically associated with cutaneous warts: HPV1, 2, 3, 4, 7, 10, 26–28, and a great number of them are associated with EV. 10,11 The problem of specific or preferential association of distinct HPVs with warts differing in morphology and/or location is still controversial, since only a limited number of cases were studied, 2–17 and no widely accepted clinical and histologic criteria are available for the differentiation between wart types. 18–21 For instance, myrmecia warts are often not distinguished from common or mosaic warts in spite of their very precise histologic and clinical description by Lyell and Miles. 22 Our aim is to present the available data on the association of different types of cutaneous warts with distinct HPVs, and to find out whether it is possible to evaluate the type of infecting HPV on the basis of clinical and histologic features. Such recognition might be important because of the differences in clinical course, contagiousness, and response to treatment of lesions associated with different HPVs.

Human papillomaviruses associated with cervical intraepithelial neoplasia. Great diversity and distinct distribution in low- and high-grade lesions.

&NA; All together, 30 genital human papillomavirus (HPV) types have been characterized so far. To evaluate the importance of HPV diversity in associated cervical diseases, we analyzed 188 biopsy specimens obtained from patients with a recent diagnosis of cervical HPV infection or intraepithelial neoplasia (CIN). Of these 188 specimens, 116 were classified as low‐grade CIN (48 cases), high‐grade CIN (53 cases), condylomata acuminata (10 cases), flat condylomas (five cases). Seventy‐two specimens were considered nondiagnostic. Using probes specific for 18 genital HPV types, HPV DNA sequences were detected by Southern blot hybridization in 100 lesions and 21 nondiagnostic specimens. When further analyzed by the polymerase chain reaction, eight HPV‐negative biopsy specimens, four CIN, and four nondiagnostic specimens were positive. Of the 129 positive biopsy specimens, 92 contained at least one of 18 known HPV types and 37 HPV that have not yet been identified. Nine specimens had more than one type. Thirteen HPV types were identified in CIN. The detection rate of HPV 16 increased from 21% in low‐grade CIN to 57% in highgrade CIN. HPV 18 was detected in only 3% of CIN; HPV 31, 33, and 35 were found in 8%. HPV 30, 39, 45, 51, 52, 56, 58, and 61 were detected in 44% of low‐grade CIN but in only 8% of high‐grade CIN. Unidentified HPV were detected in about 25% of low‐grade and high‐grade CIN. Fifty‐seven CIN positive for at least one HPV type were further analyzed by in situ hybridization. Thirty‐five (65%) biopsy specimens were positive, including 21 of 24 low‐grade CIN and 14 of 33 high‐grade CIN. Ten of the 13 previously identified HPV types were detected. Thus, CIN represents an heterogeneous disease from a virologic viewpoint. This fact could explain their variable clinical evolution.

Specificity of cytopathic effect of cutaneous human papillomaviruses

Abstract Papillomaviruses are highly tissue-specific, and are characterized by a specific mode of interaction with the squamous epithelia they infect. 1 It has been proposed that this interaction presents two successive steps that depend on the stage of differentiation of the host cell in the wart. In the basal germinal cells the viral replication is never observed. Based on the well-studied model of the papillomas induced by the Shope cottontail rabbit papillomavirus, it is most likely that the viral genome is present in the basal cells as a small number (10 to 50) of extrachromosomal copies. 2,3 The viral messenger RNAs expressed correspond to nonstructural viral proteins, as yet unidentified, but most probably involved in the altered response of basal cells to the mechanisms regulating their mitotic rate and the size of the germinal cell population, thus resulting in the formation of a papilloma. 4 Vegetative viral DNA replication is triggered upon the onset of the terminal differentiation process in the suprabasal layers, followed by the synthesis of viral structural proteins and the assembly of viral particles in more superficial layers. 1,4

Variation in the Nucleotide Sequence of Cottontail Rabbit Papillomavirus a and b Subtypes Affects Wart Regression and Malignant Transformation and Level of Viral Replication in Domestic Rabbits

ABSTRACT We previously reported the partial characterization of two cottontail rabbit papillomavirus (CRPV) subtypes with strikingly divergent E6 and E7 oncoproteins. We report now the complete nucleotide sequences of these subtypes, referred to as CRPVa4 (7,868 nucleotides) and CRPVb (7,867 nucleotides). The CRPVa4 and CRPVb genomes differed at 238 (3%) nucleotide positions, whereas CRPVa4 and the prototype CRPV differed by only 5 nucleotides. The most variable region (7% nucleotide divergence) included the long regulatory region (LRR) and the E6 and E7 genes. A mutation in the stop codon resulted in an 8-amino-acid-longer CRPVb E4 protein, and a nucleotide deletion reduced the coding capacity of the E5 gene from 101 to 25 amino acids. In domestic rabbits homozygous for a specific haplotype of the DRA and DQA genes of the major histocompatibility complex, warts induced by CRPVb DNA or a chimeric genome containing the CRPVb LRR/E6/E7 region showed an early regression, whereas warts induced by CRPVa4 or a chimeric genome containing the CRPVa4 LRR/E6/E7 region persisted and evolved into carcinomas. In contrast, most CRPVa, CRPVb, and chimeric CRPV DNA-induced warts showed no early regression in rabbits homozygous for another DRA-DQA haplotype. Little, if any, viral replication is usually observed in domestic rabbit warts. When warts induced by CRPVa and CRPVb virions and DNA were compared, the number of cells positive for viral DNA or capsid antigens was found to be greater by 1 order of magnitude for specimens induced by CRPVb. Thus, both sequence variation in the LRR/E6/E7 region and the genetic constitution of the host influence the expression of the oncogenic potential of CRPV. Furthermore, intratype variation may overcome to some extent the host restriction of CRPV replication in domestic rabbits.

Hemagglutinating activity of bovine papilloma virus

Abstract Purified preparations of bovine papilloma virus (BPV) agglutinate mouse erythrocytes. Maximal hemagglutination (HA) activity occurs at 4°, between pH 6.8 and 8.4. The adsorbed virus is readily eluted at 37°. The BPV receptors on mouse erythrocytes show a high resistance to receptor-destroying enzyme or Influenza A2 neuraminidase. The BPV hemagglutinin is associated to both full and empty viral particles. Sera of animals infected with BPV contain antibodies inhibiting the HA reaction.

Analysis of human cancers, normal tissues, and verruce plantares for DNA sequences of human papillomavirus types 1 and 2

Abstract Comparatively little is known about human papillomaviruses (HPV) because they cannot be grown in tissue culture. We have in vitro labeled DNAs from two HPVs, HPV-1 which was isolated from plantar warts, and HPV-2 which was isolated from common hand warts, and used these DNAs to examine the homology between HPV-1 and HPV-2, to examine the state of the HPV genome in papillomavirus lesions, and to assay human cancer DNAs for HPV. The specific activities of the DNAs were 5.0 × 10 7 to 1.1 × 10 8 cpm/μg. The C 0 t 1 2 of the HPV-1 and HPV-2 DNAs were 5 and 7 × 10 −4 , respectively, consistent with a genome molecular weight of about 5.2 × 10 6 . Cross-hybridization of HPV-1 and HPV-2 DNAs revealed only 5–7% homology, confirming that these are distinct viruses. HPV-1 DNA was detected by Southern blot analysis in 9 of 10 plantar warts examined. No clear evidence was found for integrated viral sequences in DNAs from eight of the nine warts analyzed. Using these HPV-1 and HPV-2 probes, we have performed the first extensive and definitive molecular hybridization analysis of human cancer DNAs for HPV sequences. Human tumor DNAs were analyzed for HPV sequences by saturation hybridization using nick-translated HPV-1 and HPV-2 DNA probes. Reconstruction experiments with added HPV-1 or HPV-2 DNAs indicated that the probes could detect 0.1 copy of the viral genome per diploid equivalent of cellular DNA. No HPV-1 sequences were detected in DNAs from 156 human cancers (14 melanoma, 3 Ca skin, 5 Ca pharynx, 1 Ca esophagus, 4 Ca stomach, 5 Ca small intestine, 22 Ca colon, 14 Ca rectum, 25 squamous cell Ca lung, 3 adenocarcinoma lung, 4 oat cell Ca lung, 21 Ca kidney, 7 Ca bladder, 3 Ca ovary, 3 Ca cervix, 4 Ca prostate, 10 non-Hodgkin lymphoma, 2 reticulum cell sarcoma [spleen]), or 27 normal human tissues (1 skin, 10 tonsil, 8 colon, 8 kidney). No HPV-2 sequences were detected in DNAs from 145 human cancers (13 melanoma, 4 Ca skin, 2 Ca pharynx, 3 Ca mouth, 7 Ca esophagus, 4 Ca stomach, 3 Ca small intestine, 29 Ca colon, 15 Ca rectum, 25 Ca kidney, 15 Ca bladder, 2 Ca ovary, 6 Ca cervix, 4 Ca prostate, 2 Ca seminoma testes, 11 non-Hodgkin lymphoma) or 1 normal human ovary. These data are strong evidence that none of the cancer specimens assayed were induced by HPV-1 or HPV-2. However, additional work is required to fully evaluate whether HPVs are possible agents of human cancers, because the cancer types assayed in this study represent only about 50% of the cancer incidence in the United States, and because our probes would not detect sequences of other recognized HPV types (HPV-3, HPV-4, and HPV-5).

The morphology of butchers' warts as related to papillomavirus types

SummaryHand warts were studied in 160 butchers. Clinical and histological studies were performed in 190 warts and virological studies in 165 warts from 104 butchers. Since we found almost perfect correlation between the histological pattern and the type of infecting virus, it was possible to evaluate the virus types in a further 39 of 56 butchers without virological studies, on the basis of the histology of the warts. The most common infection was with HPV-2 (human papilloma virus) and HPV-7. Thirty-three butchers were infected with two types of viruses and three butchers with three HPVs. The morphology of warts varied considerably. The majority were similar to verrucae vulgares or verrucae planae. Some deep warts resembled myrmecia-type verrucae plantares. Often, several types of warts coexisted. Some clinical patterns were shown to be preferentially associated with distinct types of papillomaviruses: common warts with HPV-2, HPV-4, or HPV-7, plane and intermediate warts with HPV-3, HPV-10, HPV-28. HPV-7, previously identified for the first time in these butchers, was found to be associated with common warts or common wart-like, papillomatous lesions.

Location of the T4 gene 32 protein binding site on polyoma virus DNA.

Incubation of superhelical DNA with the bacteriophage T4 gene 32 protein followed by glutaraldehyde fixation yields a circular structure with a small denaturation loop that can be visualized by electron microscopy [ 1 ] . Studying SV 40 DNA, Morrow and Berg [2,3] showed that the denaturation loop is located in a specific region (0.45 genome length) by cleavage with either one of the Eco-RI and HpaII restriction enzymes that produce unit length linears of SV 40 DNA. In a previous study, we reported (4) the presence of a single T4 gene 32 protein denaturation loop on polyoma virus superhelical DNA and its location relative to the unique Eco-Rl cleavage site. The major binding site was located at 0.22 (’ 0.02) genome lenght from the nearest Eco-Rl end and two minor sites at 0.09 (+ 0.03) and 0.41 (f 0.01). Because the two ends of the Eco-Rl generated linear molecules are indistinguishable, an ambiguity remained as to the absolute location of the T4 gene 32 protein major binding site(s) on the polyoma DNA physical map established by Griffin et al. 151. An alkaline denaturation map of polyoma (Py) DNA recently established in our laboratory [6] showed the presence of two A-T rich regions at about 0.2 and 0.8 of the polyoma DNA molecule, either of these sites or both can bind the gene 32 protein in the superhelical molecule. In an attempt to resolve this ambiguity, we searched for another restriction endonuclease that will cleave PyDNA in a unique site.

Location of the T4 gene 32 protein-binding site on polyoma virus DNA

Three easily denatured regions can be demonstrated in polyoma virus DNA. T4 gene 32 protein which binds to single stranded DNA, but not to duplex DNA, will specifically bind to any of these sites when viral DNA is in its superhelical configuration. These sites were mapped relative to a unique E. coli RI endonuclease cleavage site by electron microscopy.