There are two traditional ways to modify a virus for immunization: (1) kill the virus or (2) use a live, attenuated virus. There are three modern ways to prepare vaccines: (1) extract and purify a part of the virus that is immunogenic, (2) synthesize a polypeptide immunogen piece of the virus, or (3) use recombinant deoxyribonucleic acid or gene splicing to prepare an immunogenic portion of the virus. The last three produce subunit vaccines that can be made to contain no deoxyribonucleic acid. They are not infectious and are likely to be nononcogenic. Using recombinant deoxyribonucleic acid techniques, a vaccine for bovine papillomavirus has been prepared. This is in clinical trials and probably will be licensed for use in cattle in 1988. A vaccine for herpes simplex virus has been prepared using glycoprotein D from the surface of the virus. This immunizes animals but it has not reached clinical trials in humans.

Immunosuppressed patients such as renal transplant recipients are prone to increased incidence of wart disease. We examined 48 tissue specimens from 36 renal transplant recipients using human papillomaviruses (HPVs) 1, 2, 3, 4, 5, and 6 in filter hybridization under stringent conditions. The results showed that 90% of the samples contained HPV DNA. Of these 43 positive samples, we found HPV-1 in 2%, HPV-2 in 56%, HPV-3 in 19%, HPV-4 in 47%, HPV-5 in 9%, and HPV-6 in 5%. In several cases, more than one type of HPV DNA was observed. In a few of these cases, the clinical appearance of the lesions differed from what might have been expected, such as those lesions containing HPV-3- or HPV-5-related DNAs.

This is the report of an outbreak of human astrovirus type 5 gastroenteritis that occurred in a residential care home for older people in June 2013 in Tayside, Scotland, and which involved seven staff members and thirteen residents. This type of astrovirus has not been found in Scotland before and is rarely described in the literature.Using molecular methods such as PCR and sequencing to detect the cause of this gastroenteritis outbreak and to contain the outbreak using Public Health measures.Following an epidemiological investigation, stool samples were sent for routine virology and microbiology testing at the local microbiology and virology laboratory and were found to be negative. Further testing with real-time PCR and gene sequencing at the West of Scotland Specialist Virology Centre was performed. Data on the epidemiology and the response to the outbreak was collected.All samples had a 99% match to human astrovirus type 5. The use of standard infection control precautions with the addition of transmission-based precautions most likely contained the spread of the virus in this situation.This report illustrates the importance of using PCR and sequencing to identify pathogens such as astrovirus in outbreaks of vomiting and diarrhoea in older people particularly if routine virology and microbiology tests are negative.

Human papillomavirus (HPV) genomes were identified in two cases of verrucous carcinoma of the vagina, using Southern blot DNA hybridization under low-stringency conditions. Type (group) 6 HPV DNA (HPV-6) was identified, using molecularly cloned HPV-1 through HPV-6 DNA probes under high-stringency conditions in both cases. In addition, DNA extract in one case hybridized with HPV-1, HPV-3, and HPV-4 DNA probes. No HPV structural proteins were demonstrated in either case by immunocytochemical tests, using HPV antibodies. In one case viruslike intranuclear particles were observed by transmission electron microscopy. These two cases suggest a strong associative relationship between HPV and verrucous carcinoma (Ackerman) of the lower part of the genital tract.

Great progress has been made over the last five years in our understanding of papillomavirus (PV) biology. New technology has enabled investigators to understand the relationship between the PV and its host. The PV cannot be cultured in vitro, and this has led to limitations for those wishing to study the biology of this virus. However, utilizing recombinant DNA technology, investigators now have abundant quantities of human papillomavirus (HPV) DNA for study. Such HPV genomes may be labeled with a radioisotope such as P32 and used as a "probe" in hybridization studies to see if a given tissue contains HPV DNA. No longer are we limited to electron microscopy and immune studies in our efforts to identify HPV within benign or malignant tissues. Ultimately, we hope to understand the relationship between the virus and its host. This paper will concentrate on one aspect of this relationship--the immunology of HPV.

Benign papillomas from a patient with a family history of epidermodysplasia verruciformis were examined for the presence of human papillomavirus (HPV) DNA. Employing stringent hybridization conditions that allow identification of a single type of HPV and radioactively labeled HPV-5 DNA as a probe, we have detected HPV DNA exhibiting sequence homology to HPV-5 in these tumors. Restriction endonuclease analysis of this HPV DNA confirmed its identity as HPV type 5. However, when hybridization was performed under less stringent conditions that allow all of the known types of HPV to react with the radioactively labeled HPV-5 DNA probe, two additional species of HPV DNA unrelated to HPV-5 were identified. As these two HPV types do not hybridize with HPV 1, 2, 3, or 4 under stringent conditions, they appear unique and have, as yet, not been reported to be associated with patients exhibiting epidermodysplasia verruciformis. Thus we have observed three distinct HPV species in benign papillomas from a single patient. These observations have important implications when attempting to correlate the type of HPV present in the various wart disease syndromes that have been described to date and further suggest that extreme care must be taken when analyzing carcinomas, occupying similar anatomic sites and suspected to have arisen from papillomas, for HPV species.

We have recently identified two unusual human papillomavirus (HPV) isolates while engaged in an ongoing study of wart disease in meat handlers and veterinarians. The papillomas from which these two viruses were isolated clinically resembled verruca vulgaris rather than either flat warts or epidermodysplasia verruciformis (EV). These two previously uncharacterized HPVs were molecularly cloned and characterized with respect to known HPVs. The genomes of the two viruses exhibited dramatically different restriction endonuclease cleavage patterns but were found to have significant sequence homology to each other, as well as to HPV-3 and a new virus isolated from a patient with EV. Neither of the two new HPV isolates exhibit detectable sequence homology under stringent conditions of hybridization or share similar restriction endonuclease cleavage patterns with previously characterized HPV types 1,2,4,5,6b, or a previously isolated HPV from meat handlers.

DNA extracted from squamous cell carcinomas from patients with the chronic wart disease syndrome, epidermodysplasia verruciformis, was analyzed for the presence of human papillomavirus (HPV)-specific DNA sequences by Southern blot hybridization analysis. Employing an HPV probe obtained by molecular cloning of viral DNA purified from benign warts from these patients, we have unequivocally identified HPV-specific nucleotide sequences in squamous cell carcinomas from these patients. Restriction endonuclease mapping indicated that the DNA present in the carcinomas was of the same type (type 5) as that found in the benign tumors from these patients and was present as unintegrated, free viral DNA. Moreover, we have demonstrated the presence of HPV-5 DNA in a subcutaneous metastatic tumor from one of these patients. This latter observation essentially eliminates the possibility that the HPV-5 DNA present in the malignant tumors in these patients resulted from cross-contamination from an adjacent benign warty lesion. In addition to wild-type HPV-5 DNA, both the primary and metastatic carcinomas analyzed also contained an HPV-5 DNA species lacking approximately 20% of the HPV-5 DNA genome. These subgenomic forms of HPV-5 DNA could not be detected in benign papillomas from these patients.

The presence of papillomaviruses in epithelial-derived cancers from several animal species has led to the speculation that these viruses may also have a pathogenic role in the development of certain human carcinomas, particularly those associated with the anogenital tract. Recently, human papillomavirus (HPV) DNA has been detected in epithelial-derived cancers, both cutaneous and metastatic, from patients exhibiting the rare, chronic flat wart disease, epidermodysplasia verruciformis (EV). Except for patients exhibiting this chronic wart syndrome, the association of HPV genomes with human epithelial cancers has not been demonstrated. In an attempt to delineate the association and possible involvement of papillomaviruses with human anogenital carcinomas, we have begun an analysis of these cancers for the presence of HPV-specific nucleotide sequences by using highly sensitive hybridization procedures capable of detecting distantly related papillomaviruses at low copy number. Here we demonstrate the presence of HPV DNA in several types of anogenital tumours: Bowenoid papulosis, carcinoma in situ, and verrucous carcinoma. These data indicate that HPV can be detected in several types of premalignant and malignant tumours, supporting the contention that this group of viruses may be involved in the development of certain types of human epithelial-derived cancers.

Fifty-nine (24 plantar, 35 common) warts surgically excised from 44 patients (15 female, 29 male), 18 to 32 years old, were examined by electron microscopy (EM) for papillomavirus particles and by fluorescent antibody (FA) and peroxidase-antiperoxidase (PAP) tests for human papillomavirus type 1 (HPV-1)-specific antigens and papillomavirus genus-specific (common) antigens. Fifty per cent of plantar warts and 52 per cent of common warts were positive for HPV by EM. When examined by FA and PAP for genus-specific antigens, 58 per cent of plantar warts and 68 per cent of common warts were positive. Fifty per cent of plantar warts and 11 per cent of common warts were caused by HPV-1 as determined by reactivity with HPV-1 type-specific antiserum. All warts positive by EM were positive by FA and PAP. Sampling error accounted for warts positive by FA and PAP but not by EM. Forty-two per cent of plantar warts and 31 per cent of common warts were negative by both EM and FA and PAP. The warts caused by HPV-1 contained more virus and were more clinically aggressive than warts caused by other HPV. This study shows that the results obtained by the PAP technique using papillomavirus genus- and type-specific antisera on formalin-fixed, paraffin-embedded tissue available to the diagnostic pathologist are concordant with the results obtained by methodology usually available only to the experimental pathologist.

During the past twenty years, classic genetics has been transformed to a modem molecular science. And now we are about to witness a new step, the manipulation of genetic science to provide a technical base for industry. This new biotechnology, called recombinant DNA (rDNA) technology, is based largely upon our ability to identify specific genetic information and to transfer genetic information from one organism or cell to another. Perhaps our greatest exposure to rDNA is the pages of The Wall Street Journal touting the commercial impact upon business and investment, but it is important to focus for a moment upon the science and the potential effect upon our lives and those of our patients. Most readers have a working knowledge of rnendelian genetics and the rudiments of nucleic acid chemistry. However, few of us followed the advances leading to the discipline of rDNA or gene splicing. In the 1960s, scientists learned how genes moved from one bacterium to another by the transfer of virus-like bacteriophages or separate small circular DNA plasmids. Thus, phages and plasmids were identified as vectors for carrying genetic information. Soon investigators learned how bacteria have the capacity to clip genetic information from chromosomes and repair the resultant breaks. Over 200 bacterial enzymes have now been identified, each with a specific ability to cut and repair DNA. The final step in this process involved understanding the mechanisms of gene expression, including the identification of nucleic acid sequences that initiate and terminate protein synthesis. Put this all together and we have rDNA technology: the capacity to select a gene of specific function and transfer the gene via a vector to bacteria where large

Papillomavirus DNA from common skin warts of four meathandlers has been characterized and compared to the DNA genomes of known human cutaneous papillomaviruses. Virus was purified from one of the human papilloma samples and was found to exhibit a restriction endonuclease cleavage pattern distinct from those of known human papillomaviruses (HPV). Moreover, viral DNA extracted from papillomas from the other three patients exhibited sequence homology and restriction endonuclease cleavage patterns similar to this novel viral DNA. In addition to this unique HPV, one of the patients also contained a second species of HPV which was related to, yet distinct from, HPV-2, a species of HPV associated with common cutaneous warts in man. Finally, employing the Southern transfer procedure and stringent hybridization conditions, each meathandler HPV DNA studied was found to be unrelated to the major species of bovine papillomavirus, types 1 and 2.

Anogenital warts from 26 patients were examined for the presence of human papillomavirus (HPV). Although no whole, intact virus could be identified, varying amounts of nonintegrated HPV DNA were detected in 18 tissue specimens (70%) by employing both an agarose gel-ethidium bromide staining method and the Southern blot hybridization procedure. When hybridization analysis was performed under stringent conditions, six anogenital warts were observed to contain HPV genomic sequences related to either of the cutaneous viruses HPV type 1 (HPV-1) or HPV-2. In 12 tissue samples lacking sequence homology to either HPV-1 or HPV-2 under stringent conditions, HPV-related sequences were detected when the hybridization was performed under less stringent conditions, indicating that an HPV distinct from both HPV-1 and HPV-2 is also associated with these lesions. This anogenital HPV also appeared to be distinct from the other characterized types of HPV. These data indicate that at least three HPVs are associated with anogenital wart disease.

Paraffin sections of condylomata acuminata removed from the lower genital canal were stained for papillomavirus antigen by th peroxidase-antiperoxidase test using a broadly cross-reactive antiserum. The antiserum was prepared by immunization of a rabbit with disrupted capsids of papillomavirus purified from a pool of plantar warts. Specific staining was seen as a brown granular reaction in the nuclei of the epithelial cells; this reaction occurred most consistently in the more superficial cells. Papillomavirus antigen was demonstrable in about half of the 50 specimens examined. The antigen was found in both flat and papillary lesions from the vulva, vagina, and cervix. The distribution of the antigen was widely variable and ranged from abundant in some specimens to patchy and sparse in others. In papillary lesions, antigen-positive cells were found characteristically at the tips of the epithelial fronds. The ability to detect the viral antigen in genital condylomas may help in understanding the pathogenesis of these lesions and in evaluating the role of papillomaviruses in th etiology of lower genital tract cancer.

Annals of the New York Academy of SciencesVolume 354, Issue 1 p. 60-79 GENETIC VARIATION AMONG PAPILLOMAVIRUSES* Anthony J. Faras, Anthony J. Faras Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorRichard A. Krzyzek, Richard A. Krzyzek Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Supported by the National Institutes of Health (Young Investigators Award CA 23833).Search for more papers by this authorRonald S. Ostrow, Ronald S. Ostrow Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorSusan L. Watts, Susan L. Watts Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Supported by the National Institutes of Health (Training Grant 09138).Search for more papers by this authorDonna M. Smith, Donna M. Smith Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorDwight L. Anderson, Dwight L. Anderson Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorCedric A. Quick, Cedric A. Quick Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorFranklin Pass, Franklin Pass Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Departments of Microbiology and Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this author Anthony J. Faras, Anthony J. Faras Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorRichard A. Krzyzek, Richard A. Krzyzek Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Supported by the National Institutes of Health (Young Investigators Award CA 23833).Search for more papers by this authorRonald S. Ostrow, Ronald S. Ostrow Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorSusan L. Watts, Susan L. Watts Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Supported by the National Institutes of Health (Training Grant 09138).Search for more papers by this authorDonna M. Smith, Donna M. Smith Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorDwight L. Anderson, Dwight L. Anderson Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorCedric A. Quick, Cedric A. Quick Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this authorFranklin Pass, Franklin Pass Departments of Microbiology and §Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455 Departments of Microbiology and Dermatology University of Minnesota Medical School Minneapolis, Minnesota 55455Search for more papers by this author First published: November 1980 https://doi.org/10.1111/j.1749-6632.1980.tb27958.xCitations: 11 * Supported by the National Institutes of Health (Grants CA 25462 and CA 25124) and the Minnesota Leukemia Research Foundation. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume354, Issue1Genetic Variation of VirusesNovember 1980Pages 60-79 RelatedInformation

An antiserum prepared by immunization of a rabbit with sodium dodecyl sulfate-disrupted virions from a pool of plantar warts was cross-reactive with virus-positive papillomas of other animal species by both indirect immunofluorescence tests on frozen sections of wart tissues and peroxidase-antiperoxidase tests of sections of Formalin-fixed tissues. The antiserum stained plantar warts, common warts, and skin lesions of epidermodysplasia verruciformis, all from humans; bovine fibropapilloma, experimentally produced with bovine types 1 and 2; and transmissible canine oral papillomas. The staining was localized to nuclei of the upper granular layers of the peithelium and was similar in distribution to the pattern produced by antiserum specifically prepared against that papillomavirus. The antiserum did not stain virus-negative warts, or cells infected with simlan virus 40, human polyomavirus BK, and murine polyomavirus. These data suggested that papillomaviruses share a common internal antigen unrelated to a similar antigen described previously for the polyomaviruses (which include simian virus 40 and polyomavirus subgroups).