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G Orth

Publications and source records attributed to G Orth.

At least 127 records · Page 7Linked to original sources

An immunofluorescence complement-fixation test for detection of human papilloma viruses in various warts and wartlike lesions of epidermodysplasia verruciformis.

Comparative studies of indirect immunofluorescence (IF) and IF complement fixation tests were performed in 11 warts induced by various human papilloma viruses (HPV) or wartlike lesions of epidermodysplasia verruciformis. For the detection of HPVs, specific immune sera against HPV1, HPV2, HPV3, HPV5, HPV9 and/or HPV8 were used. The complement-fixation test proved to be as specific as the indirect IF method for the detection of HPVs in the tissues and was superior in clarity of IF readings (no nonspecific IF staining).

Complement Fixation Tests↗

Introduction of cloned human papillomavirus genomes into mouse cells and expression at the RNA level.

The entire DNA genomes of five different human papillomaviruses (HPVs) were cloned into the BamHI site of pBR322 (HPV-1a, HPV-3, HPV-4, and HPV-9) or the EcoRI site of pBR325 (HPV-2), using as starting materials virus preparations isolated from papillomas of individual patients. Under stringent hybridization conditions (Tm-28 degrees), the five cloned HPVs exhibited less than 10% homology with one another. To establish model cell systems that may be useful for the identification of HPV genes and HPV gene products, mouse thymidine kinase negative (tk-) cells were cotransformed to the tk+ phenotype with the herpesvirus thymidine kinase gene and each of the five HPV cloned DNAs (either as intact recombinants or excised HPV DNA without removal of pBR). In most tk+ cell clones, a complex pattern of multiple high molecular weight inserts of HPV DNA were present in high copy number. Most of the HPV DNA sequences in the cotransformed cells were not present as unit-length episomal viral DNA. Analyses of the integration pattern (DNA blot) and RNA expression (RNA blot) of several HPV-1a and HPV-3 transformed cell lines suggest that some copies of the viral genome are integrated in a similar manner in different cell lines leading to the expression of identical viral RNA-containing species. Two of the cell lines transformed by the intact HPV-1a/pBR322 recombinant synthesized substantial amounts of four discrete viral polyadenylated cytoplasmic RNA species of 1.9, 3.2, 3.8, and 4.5 kb. Two cell lines transformed by the intact HPV-3/pBR322 recombinant synthesized 4-5 polyadenylated cytoplasmic viral RNA species ranging from 0.8 to 4.6 kb. The analysis shows that each viral RNA species appears to be a hybrid RNA molecule containing both HPV and pBR322 sequences. Based on these findings and the molecular organization of the HPV-1a genome (O. Danos, M. Katinka, and M. Yaniv (1982). EMBO J. 1, 231-237), it is possible that transcription of each of the HPV-1a RNA species is initiated using the HPV early promoter and terminated in pBR322.

Animals↗

Bovine papillomavirus type 1 genome in hamster sarcoma cells in vivo and in vitro: variation in the level of transcription.

The physical state and expression of the bovine papillomavirus type 1 (BPV-1) genome were analysed in two transplantable hamster sarcomas (HT1 and HT2) after a low number of transplantations, in two tumourigenic cell lines derived from the first transplant of HT2 sarcoma and another transplantable sarcoma (HT3) and in tumours obtained by grafting these cells. Blot hybridization experiments indicated the presence of multiple free copies of the whole viral genome in HT1 and HT2 tumour transplants (100 and 25 copies/cell respectively), irrespective of the number of in vivo or in vitro passages. In contrast, HT3 cells, and tumours induced by these cells, at early and late passages in vitro contained viral sequences probably integrated in the cell genome, in addition to the free viral DNA sequences. Polyadenylated viral transcripts were easily detected in HT1, HT2 and HT3 tumours obtained at early passages in vivo and in vitro, with electrophoretic mobilities corresponding to 1200 to 1400 bases (HT1, HT2 and HT3), 1750 bases (HT1) and 2500 bases (HT2). Homologous sequences of the transcripts were localized in the transforming BamHI-HindIII fragment of BPV-1 DNA, mainly in the BamHI-EcoRI fragment (0.31 to 0.602 map units). In contrast, almost no viral transcription was detected in HT2 and HT3 cells after 50 subcultures and in the tumours induced by these cells. This suggests that the tumourigenicity of the HT2 and HT3 cells is compatible with a very low level of expression of the transforming region of the BPV-1 genome.

Animals↗

Humoral and cell-mediated immunity to human papillomavirus type 1 (HPV-1) in human warts.

The humoral and cell-mediated immune response to human papillomavirus type 1 (HPV-1) has been studied in 162 patients carrying papillomas of various clinical types: deep plantar wart or myrmecia, common wart, flat wart, and anogenital wart. Circulating antibodies were detected by immunodiffusion and microcomplement fixation, using purified HPV-1 particles as type-specific antigen. A significant association between myrmecia and anti-HPV-1 antibodies was found (39% of the cases). Cell-mediated immunity was evaluated by a study of delayed hypersensitivity (DH). The main capsid components of HPV-1 (HPV-1 CP), consisting mostly of a polypeptide of molecular weight 54,000, were injected intradermally. In addition to the type-specific antigens, HPV-1 CP contain other antigenic determinants shared by various types of human papilloma-viruses and masked in intact viral particles. The DH tests to HPV-1 CP showed no differences between the carriers of different papilloma types, confirming the presence of common antigenic determinants. Moreover, they gave rise to an increase or to new anti-HPV-1 antibody production mostly in myrmecia carriers (78% and 33% of the cases, respectively), and to new DH to HPV-1 CP in all groups of papilloma carriers (33% to 56%, depending on the clinical papilloma type).

Adolescent↗

Human papillomaviruses associated with epidermodysplasia verruciformis. II. Molecular cloning and biochemical characterization of human papillomavirus 3a, 8, 10, and 12 genomes.

The DNAs of four human papillomaviruses (HPVs) that were found in the benign lesions of three patients suffering from epidermodysplasia verruciformis have been characterized. The flat wart-like lesions and the macular lesions of patient 1 contained two viruses, HPV-3a and HPV-8, respectively, whose genomes had previously been only partially characterized. The flat wart-like lesions of patient 2 and the macular lesions of patient 3 each contained a virus previously considered as belonging to types 3 and 5, respectively. These viruses are shown in the present study to be different from all of the HPV types so far characterized; they have tentatively been named HPV-10 and HPV-12. The HPV-3a, HPV-8, and HPV-12 DNAs and the two SalI fragments of HPV-10 DNA (94.1 and 5.9% of the genome length) were cloned in Escherichia coli after having been inserted in plasmid pBR322. The cloned HPV genomes have similar sizes (about 7,700 base pairs), but their guanine-plus-cytosine contents differ from 41.8% for HPV-12 DNA to 45.5% for HPV-3a DNA. The study of the sensitivity of the four HPV DNAs to 14 restriction endonucleases permitted the construction of cleavage maps. Evidence for conserved restriction sites was found only for the HPV-3a and HPV-10 genomes since 5 of the 21 restriction sites localized in the HPV-3a DNA seem to be present also in the HPV-10 DNA. Hybridization experiments, performed in liquid phase at saturation, showed a 35% sequence homology between HPV-3a and HPV-10 DNAs, 17 to 29% sequence homology among HPV-5, HPV-8, and HPV-12 DNAs, almost no sequence homology between the HPV-3a or HPV-10 DNA and the other HPV DNAs, and a weak homology between HPV-9 DNA and HPV-8 or HPV-12 DNA. Blot hybridization experiments showed no sequence homology between the HPV-3a, HPV-8, and HPV-12 DNAs and the DNAs of the HPVs associated with skin warts (HPV-1a, HPV-2, HPV-4, and HPV-7) or with mucocutaneous and mucous membrane lesions (HPV-6b and HPV-11a, respectively). One exception was a weak sequence homology between the HPV-2 prototype and HPV-3a or HPV-10 DNA.(ABSTRACT TRUNCATED AT 400 WORDS)

Base Composition↗

Oncogenic potential of human papillomaviruses epidermodysplasia verruciformis: a counterpart of Shope papilloma-carcinoma complex.

Epidermodysplasia verruciformis, a model of viral oncogenesis in humans, shows a remarkable similarity to Shope papilloma-carcinoma complex in rabbits. In humans and rabbits, both host, i.e. genetics and immunity, and environmental factors are decisive for neoplastic conversion of benign proliferative lesions. The virus seems to function as an initiation and promotor of carcinomatous transformation. In epidermodysplasia verruciformis, specific virus types are involved in malignant conversion, but a deeply depressed cell mediated immunity is responsible for infection with the potentially oncogenic viruses.

Animals↗

[Condylomatous lesions of the uterine cervix: their course in 2466 patients].

2466 women with cervical condylomatous lesions out of the general consulting (0.7%) have been followed by the authors. The cytological and histological criteria of these lesions and the detection of the viral antigen by immunoperoxidase (positive in only about 50% of the cases), are recalled. The flat condylomas are often associated with dysplasia (CIN I, II, III). The condylomas appear in women before the age of 20. These cases increased in number between the ages of 25 to 30 and stayed high until 35. The number of condylomas associated with CIN II have their maximum between 36 and 38 years of age and decrease afterwards to age 48. The graphic is the same for CIN III. The evolution of these condylomatous lesions studied during 42 months, shows that in 1269 women with condyloma and nuclear atypia, regression occurred in 53 per cent, persistence in 37 per cent and aggravation in 10 per cent of the cases. In 762 women with CIN II, regression appeared in 39 per cent, persistence in 44 per cent and aggravation in 17 per cent of cases. In a group of 764 closely followed women, regression and aggravation in CIN I and II appeared between the 3rd and the 6th months of follow-up. Condyloma associated with CIN III were not observed after the 3rd month. Recurrence appeared however in 55 cases after insufficient ablation. Lastly, the histogenesis of these lesions and the relationship between the viral action and the host are discussed.

Adolescent↗

[Detection and mapping of conserved nucleotidic sequences between the genomes of human papillomavirus 1 a and bovine papillomavirus 1 by electron microscope heteroduplex analysis (author's transl)].

Three regions of partial homology have been detected between the genomes of human papillomavirus 1 a (HPV-1 a) and bovine papillomavirus 1 (BPV-1) by electron microscope analysis of heteroduplex molecules. These regions contain about 25% of non-homologous bases and represent 13% of the genome length. They have been mapped on the viral DNAs. This allowed a reciprocal orientation of the physical maps of the two genomes. Two regions are located in the middle of the transforming fragment of BPV-1 DNA and the third, at at diametrically opposite position, falls into the region of the genome coding most probably for a viral structural protein.

Animals↗

Two anatomoclinical types of warts with plantar localization: specific cytopathogenic effects of papillomavirus. Type I (HPV-1) and type 2 (HPV-2).

In this study, the clinical and histopathological aspects of 50 plantar warts are reported in relation to the type of papillomavirus present in the lesions, as detected by immunofluorescence tests, using specific guinea pig fluorescein-labelled IgG. Warts of plantar localization are not caused by the same human papillomavirus (HPV) since they are found to be associated with both HPV type 1 (HPV-1) and HPV type 2 (HPV-2). HPV-1 is always associated with deep and painful plantar warts (myrmecia), whereas HPV-2 is found to be associated with superficial, painless plantar warts (vulgaris or often mosaic type). Histologically, these two types of plantar warts are quite different. In myrmecia (HPV-1), characterized by an endophytic growth, large eosinophilic, keratohyaline-like granules are observed in the cytoplasm and nucleus of infected, often vacuolated cells. These granules appear early in stratum spinosum and are very numerous in stratum granulosum. In the mosaic type (HPV-2), the histopathological aspect is not different from that of common warts; these lesions have an exophytic growth and are characterized by foci of clear vacuolized cells which are found in stratum granulosum. Their cytoplasm contains round, basophilic keratohyalin granules which often have a heterogenous aspect. These differences are observed in other localizations of morphologically related warts associated with HPV-1 and HPV-2 and seem to be related to a specific cytopathogenic effect of HPV-1 an HPV-2 in human papillomas.

Cytopathogenic Effect, Viral↗

Biochemical characterization of two types of human papillomaviruses associated with epidermodysplasia verruciformis.

The DNAs of the human papillomaviruses (HPVs) associated with the benign lesions of two patients suffering from epidermodysplasia verruciformis (patients JD and JK) were analyzed by using 12 restriction endonucleases. None of the restriction endonucleases were one-cut enzymes for the HPV DNA obtained from patient JD, referred to as the prototypical HPV-5, whereas five of them were one-cut enzymes for the DNA of the major virus found in patient JK, referred to as HPV-9. The molecular cloning of the two fragments resulting from the cleavage of HPV-5 DNA by endonuclease HindIII and of the single fragment obtained after treatment of HPV-9 DNA with endonuclease BamHI was performed in Escherichia coli after the fragments were inserted in plasmid pBR322. A cleavage map of the two cloned genomes was constructed. Little sequence homology (4 to 5%) was detected between HPV-5 and HPV-9 DNAs by DNA-DNA hybridization experiments in liquid phase at saturation; this homology was reproducibly higher than that (2 to 3%) detected under the same conditions between these DNAs and HPV-1a DNA. In addition, blot hybridization experiments performed under stringent conditions showed no or little sequence homology between the DNAs of HPV-5 and HPV-9 and those of HPV prototypes of types 1, 2, 3, 4, and 7 associated with skin warts. These results confirm that HPV-5 and HPV-9 are two distinct HPV types.

Animals↗