Biomedical subjects
Publications and source records attributed to H Pfister.
Human papillomavirus (HPV) DNA was isolated from a clinically diagnosed flat wart and proved to be related to HPV2. The isolate showed 55% cross-hybridization with HPV2a. A physical map of restriction enzyme cleavage sites differed completely from those of HPV2a and HPV2b. The new HPV2 subtype, which will be classified as HPV2c, was found to be very prevalent in common warts.
2 cases of acanthosis nigricans associated with an adenocarcinoma (signet ring cell carcinoma) of the stomach and a metastasizing small-cell carcinoma of unknown origin are reported. In both cases the skin lesions preceded the diagnosis of the carcinoma by months and acanthosis nigricans maligna was suspected by onset and localization of the dermatosis. There was no evidence of a papillomavirus etiology of the warty skin lesions. Virus particles could not be demonstrated either in ultrathin sections or in buffer extracts. Virus-specific DNA was not detectable after CsCL-ethidium bromide gradient centrifugation and cellular DNA did not hybridize under stringent or relaxed conditions with 32P-labelled human papillomavirus 6 or human papillomavirus 8 DNA.
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Albumin and IgG concentrations were determined in the cerebrospinal fluid (CSF) and serum from 21 normal dogs and 79 dogs with neurological disease, using rocket immunoelectrophoresis. A qualitative protein reaction, cell count and cytological examination were also performed on most CSF samples. The values obtained in normal dogs were used to establish baseline data. Those obtained in neurologically ill dogs were divided into five groups according to the clinical and pathological findings; infectious inflammatory diseases, tumours, degenerative conditions, spinal cord compression and vascular disorders. The absolute CSF albumin content was used as an indicator for blood-brain barrier disturbance and the IgG index for the detection of intrathecal IgG synthesis. Four categories of findings were distinguished; normal CSF, blood-brain barrier disturbance (increased transudation), intrathecal IgG synthesis, combined barrier disturbance and IgG synthesis. The results correlated largely with the pathological findings although in some cases the CSF values could not be explained by the morphological data. It was concluded that rocket immunoelectrophoresis, although it is an elaborate technique, is a useful addition to other CSF examination methods.
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The 7.2 kb genomic DNA of bovine papillomavirus type 3 (BPV-3) was molecularly cloned using its unique EcoRI site, and the 7.3 kb genome of BPV-4 was cloned using its single BamHI site. The viral genomes were compared by liquid hybridization, Southern blot hybridization and heteroduplex mapping. Low stringency hybridization conditions revealed that the genomes are colinear but the sequences are extensively mismatched. The relative alignment of the restriction endonuclease maps of the two viral genomes has been determined. It was found that the genomes as linearized for cloning are out of phase by 1.7 kb, so that the single EcoRI site of BPV-3 appears to coincide with the BPV-4 EcoRI site at 0.22 map units. It is concluded that the genomes of BPV-3 and BPV-4, both of which cause true epithelial warts, share the same physical organization but exhibit sequence divergence.
Focal epithelial hyperplasia Heck lesions of a Turkish patient were shown to contain papillomavirus-specific DNA, which was molecularly cloned into bacteriophage lambda. It proved to be related to human papillomavirus (HPV) type 6 DNA and HPV type 11 DNA. Reassociation kinetics revealed a cross-hybridization of 4 and 3%, respectively. There was no cross-reactivity with HPV type 1, 2, 3, 4, 5, 8, or 10. This papillomavirus type will be referred to as HPV type 13. The DNA was characterized by cleavage with several restriction enzymes, and the cleavage sites were physically mapped. Papules from two additional cases of Morbus Heck contained HPV type 13 DNA as shown by Southern blot hybridization and by the characteristic cleavage patterns. This may indicate that HPV type 13 is more frequently associated with focal epithelial hyperplasia Heck than are other HPV types.
Persisting human papilloma virus (HPV) 2 induced common warts of a chronic lymphatic leukemia patient were orally treated with aromatic retinoid Ro 10-9359 (Tigason). Clinically, the lesions improved rapidly. Virus-specific cytopathogenic effects (CPE), virus particles and viral DNA were no longer detectable. Therapy was discontinued because of the development of a liposarcoma, which led to a complete relapse of the cutaneous lesions. HPV-2 specific parameters, CPE and viral DNA, were restored. Comparison of the restriction enzyme cleavage patterns revealed that warts before and after therapy contained the same HPV-2 subtype. The implications of the observation on the effect of Tigason on virus-induced papillomas are discussed.
An epidermodysplasia verruciformis patient suffering from generalized warts and a carcinoma at the forehead was found to be infected by at least six types and subtypes of human papillomaviruses. The central part of the carcinoma, however, harbored only human papillomavirus 5 DNA. The DNA persisted extrachromosomally in high genome copy number. In contrast to wart DNA preparations, a significant part of the viral sequences in the carcinoma was present as oligomers, at least part of them being concatemers as shown by S1 digest. The human papillomavirus 5 subtype from this carcinoma was compared with the two other carcinoma isolates described so far and proved to be rather similar.
Several antigens including immunoglobulin light and heavy chains, C3, macrophage enzymes and various brain proteins were demonstrated immunohistologically in paraffin sections of canine tissues. The indirect immunoperoxidase, the unlabelled antibody enzyme (PAP), double bridging PAP, and biotinavidin-peroxidase (BAP) methods were compared. The influence of various histological procedures such as fixation and embedding, and other modifications such as enzyme treatment of sections was investigated. The best results were obtained with the PAP and BAP methods on formalin fixed tissues. Trypsin proved to be highly effective for demonstrating certain antigens but required firm adhesion of the sections. Prolonged incubation with the primary antisera improved the results considerably. Based on the results of this study a general strategy for solving immunohistological problems has been proposed.
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Forty warts from different patients and of different clinical type were examined histologically and virologically. Eight lesions were found to be associated with human papillomavirus type 1 (HPV 1), 15 tumors were induced by HPV 2, HPV 3 was detected 4 times, HPV 4 twice, and HPV 6 eleven times. HPV 3, HPV 4, and HPV 6 induced warts revealed a correlation between histology and virus type. They are characterized by the so called "edematous type clear cells". In HPV 3 associated flat warts pycnotic nuclei were mainly localized in the center of large vacuoles. In genital warts sickle shaped nuclei were pushed to the margin of the vacuolized cells. The histology of HPV 1 and HPV 2 induced warts was more heterogenous. With one exception HPV 1-induced lesions represented typical myrmecia warts, varying in the number and shape of inclusion bodies. HPV 2 associated common warts, however, revealed 3 very distinct histologic features: (1) Inclusion wart typical for HPV 1, (2) Classical common wart with marked condensation of keratohyalin granules, (3) Warts with extreme vacuolization of squamous and granular cells leading to a honeycomb-like picture.
Three patients with focal epithelial hyperplasia Heck are presented. In two cases biopsies were made and investigated biochemically and by electron microscopy. In one case typical papillomavirus and HPV 1-DNA could be demonstrated, in the other the attempts of isolation failed. Virologic results and epidemiologic considerations suggest a viral etiology of focal epithelial hyperplasia Heck, but they are not sufficient to prove this.
A case of epidermodysplasia verruciformis in a patient from Upper-Volta is described. Slightly elevated, flat warts were observed on hands, feet, arms and legs, and pityriasis versicolor-like lesions were found mainly on the trunk. The patient showed no malignant tumors. Histological examination revealed hyperkeratosis, granulosis and moderate acanthosis with large, foamy, basophilic keratinocytes in stratum granulosum and stratum spinosum. Papillomavirus particles could be prepared from these lesions and were differentiated from known papillomavirus types by immune electron microscopy with monospecific antisera and by DNA-DNA hybridization. The viral DNA was characterized by cleavage with several restriction endonucleases and a physical map of the resulting fragments was established. The virus is designated as HPV 8. Preliminary seroepidemiologic studies with human sera indicate a rather wide distribution of HPV 8. Blot hybridization of DNA from human carcinomas with 32P-labelled virus DNA detected no HPV 8-specific sequences.
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