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BACKGROUND Testing for the presence of the human papillomavirus (HPV) is widely accepted for triaging Papanicolaou cytology results categorized as atypical squamous cells of undetermined significance (ASC-US). In contrast, HPV testing has limited use in triaging cytological low-grade squamous intraepithelial lesions (LSILs) due to prevalence rates of typically >80%. In the current study, the authors assessed the diagnostic performance of p16/Ki-67 dual-stained cytology in triaging ASC-US and LSIL cases within the prospective, multicentric Primary ASC-US LSIL Marker Study (PALMS). METHODS A total of 575 ASC-US cases and 529 LSIL cases from a cohort of 27,349 women who were prospectively enrolled into the PALMS study in 5 European countries were tested with p16/Ki-67 dual-stained cytology and Hybrid Capture 2 (HC2) HPV testing. Colposcopy-guided biopsy results of cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) were used as clinical endpoints. RESULTS p16/Ki-67 dual-stained cytology demonstrated comparable (ASC-US: 94.4% for dual-stained cytology vs 100% for HC2 testing; P = .317) or lower (LSIL: 85.7% for dual-stained cytology vs 98.4% for HC2 testing; P = .005) sensitivity for CIN2+, but higher levels of specificity compared with HC2 HPV testing in both ASC-US (78.7% vs 60.4%; P

Computerassistenz kann Screeningfehler reduzieren. Fur die Zervixzytologie gibt es nur 2 Systeme: Das BD FocalPoint™ GS Imaging System (BD) verwendet Bilderkennungsalgorithmen, um Dunnschichtpraparate (Surepath, SP) und konventionelle Abstriche auszuwerten. Das ThinPrep® Imaging System basiert auf der densitometrischen Messung von Zellkernen. Diese Technik erfordert eine Dunnschichtpraparation. BD FocalPoint™ teilt die Praparate mit absteigender Wahrscheinlichkeit fur das Vorliegen einer Auffalligkeit in 6 Gruppen ein. Eine weitere Option ist das „location guided screening“ (LGS). Hierbei werden an einem computerisierten Mikroskop die 15 Gesichtsfelder prasentiert, die mit hochster Wahrscheinlichkeit relevant sind. Mehrere Studien weisen darauf hin, dass mindestens eine Aquivalenz dieses Modus zum manuellen Screening bei konventionellen und SP-Praparaten besteht. Beim ThinPrep® Imaging System, das nur mit einem LGS-Modus arbeitet, fand die Mehrzahl der vorliegenden Studien eine signifikant hohere Sensitivitat fur histologisch bestatigte zervikale intraepitheliale Neoplasien vom Grad CIN 2 +. Allerdings hatten die meisten Studien ein retrospektives bzw. paralleles Design. Beide Systeme ermoglichen eine deutliche hohere Produktivitat, setzen allerdings auch einen hoheren technischen Gesamtaufwand voraus.

Persistent infection with HPV 16 and 18 has been causally associated with the development of cervical cancer and its precursor lesions as well as with other carcinomas and their precursors, e.g. some vulvar and vaginal cancers. Furthermore HPV 6 and 11 are responsible for anogenital condylomata acuminata in more than 90% of cases. With the recently developed prophylactic bivalent (HPV 16 and 18) and quadrivalent (HPV 6, 11, 16 and 18) vaccines, it is possible to prevent infection of the cervical epithelium and other squamous epithelia, the development of premalignant lesions and, in the case of the quadrivalent vaccine, the development of condylomata acuminata. The following paper represents a summary of the full-text version of the German evidence-based Guidelines, including all evidence-based recommendations regarding the safety as well as the efficacy of the vaccines in preventing CIN, VIN/VaIN, genital warts and other HPV-associated lesions.

Demonstration of human papilloma virus DNA (HPV-DNA) in the lesions of focal epithelial hyperplasia in an 11-year-old Turkish girl provides further proof of its viral cause. In this case characterization of HPV-DNA showed a reaction with that of type 13 and 18. This previously undescribed variant is probably the expression of a new HPV type.

Persistent condylomata acuminata in a 21-year-old patient with diabetes mellitus were treated with highly purified interferon-alpha (IFN-alpha) obtained by recombinant DNA technology. Daily dose was 1.5 X 10(6) IU, given subcutaneously. Already during treatment the condylomata regressed. Two weeks after the end of therapy, i.e. after a total dose of 10.5 X 10(6) IU IFN-alpha, all condylomata had completely receded. Blood glucose levels remained constant with concomitant insulin therapy. Toxic side-effects or antibodies to IFN-alpha were not observed.

OBJECTIVE: Overexpression of p16 INK4a has been proposed as a biomarker helpful for the identification of dysplastic cervical epithelial cells on histologic slides as well as in cervical smears. Since a few nontransformed cells in the genital tract in some instances may also express p16 INK4a , we evaluated whether applying established morphologic criteria for cervical dysplasia allows a distinction of dysplastic from nondysplastic p16 INK4a -stained cells in cytologic samples. STUDY DESIGN: Liquid-based cytology samples were obtained from a screening population (n = 50), and from patients attending a dysplasia clinic (n = 40). Slides prepared from these samples were stained with the conventional Papanicolaou stain procedure. From each specimen, a second slide was prepared in parallel and immunostained for p16 INK4a . Cytologic diagnoses for most patients attending the dysplasia clinic could be compared to the reported histologic diagnoses on punch biopsy samples taken from the patients at the time of colposcopy. This allowed a comparison of the cytology and p16 INK4a immunostaining results with subsequent hematoxylin and eosin- based histologic diagnoses. RESULTS: Overall, in 10% of slides obtained from patients with nonsuspicious smears, few p16 INK4a -positive cells were found. Using established morphologic criteria and applying these criteria on cells showing any p16 INK4a immunoreactivity, p16 INK4a -positive normal or metaplastic cells could be discriminated from p16 INK4a -expressing dysplastic cells. In 21 of 22 cases (95%) of high grade lesions (cervical intraepithelial neoplasia 2 or higher in follow-up histology), easily recognizable p16 INK4a -positive dysplastic cells could be detected, with the remaining case lacking dysplastic cells in the thin-layer slide used for p16 INK4a immunostaining. CONCLUSION: Established morphologic criteria for cervical dysplasia can be readily applied to p16 INK4a -immunostatined cytologic specimens. Thus, p16 INK4a immunostaining may help to avoid ambiguities in the interpretation of cervical cytology samples and facilitate more rapid diagnosis and possibly even automated screening of cytologic slides.

Die Sensitivitat einer HPV-Testung fur hohergradige Praneoplasien und Neoplasien der Zervix ist extrem hoch. Daher kann sie, optimal in Verbindung mit der Dunnschichtzytologie, zur Abklarung grenzwertiger, schwer beurteilbarer und auffalliger zytologischer Befunde eingesetzt werden. Bei Pap IIW, erstmaligem Pap IIID und Pap III ohne Verdacht auf glandulare Atypie ist eine primare HPV-Testung sinnvoll. Bei HPV-Negativitat kann bei Pap IIW sofort, bei Pap IIID und Pap III ohne Verdacht auf glandulare Atypie nach 2 unauffalligen Befunden eine Ruckkehr zum normalen Vorsorgeintervall erfolgen. Bei wiederholtem Pap IIID, Pap III mit Verdacht auf glandulare Atypie und Pap IVa sollte primar eine Differenzialkolposkopie mit Biopsie eventueller Herdbefunde durchgefuhrt werden. Bei fehlendem Nachweis solcher Herdbefunde muss bei Pap III mit Verdacht auf glandulare Atypie und bei Pap IVa (nach Uberprufung der zytologischen und kolposkopischen Befunde) eine Konisation (moglichst LLETZ) vorgenommen werden. Bei einem HR-HPV-positiven Pap IIID kann unter bestimmten Umstanden in 3-monatigen Abstanden kontrolliert werden.

Humane Papillomviren (HPV) sind der wesentliche Faktor bei der Entstehung des Zervixkarzinoms und seiner Vorstufen. Bei 99,7% aller invasiven Zervixkarzinome aus 22 verschiedenen Landern konnte HPV-DNA nachgewiesen werden [26]. Zahlreiche In-vitro- und In-vivo-Studien zeigen das transformierende Potential dieser Viren. Die Onkogene E6 und E7 der HPV-Typen 16 und 18 wurden bereits 1995 von der IARC als Karzinogene eingestuft. Dies gilt nach neuesten Erkenntnissen auch fur die die E6- und E7-Gene weiterer sog. Hochrisiko-HPV-Typen [28]. Eine persistierende Infektion mit Hochrisiko-HPV ist notwendige Voraussetzung fur die Entwicklung einer zervikalen intraepithelialen Neoplasie (CIN) III [4, 15]. Jedoch sind Kofaktoren fur dieses Geschehen unerlasslich. Diese sind noch unzureichend bekannt. Sicher ist jedoch, dass einer Immunsuppression, dem Rauchen und weiteren genitalen Infektionen (am besten definiert fur Chlamydien) Bedeutung zukommt. Mittlerweile gilt der Wert einer HPV-Testung sowohl im Screening als auch in der Beurteilung von Patientinnen mit auffalligen zytologischen Abstrichen als gesichert [15].

Humane Papillomviren (HPV) sind der wesentliche Faktor bei der Entstehung des Zervixkarzinoms und seiner Vorstufen. Bei 99,7% aller invasiven Zervixkarzinome aus 22 verschiedenen Landern konnte HPV-DNA nachgewiesen werden [26]. Zahlreiche In-vitro- und In-vivo-Studien zeigen das transformierende Potential dieser Viren. Die Onkogene E6 und E7 der HPV-Typen 16 und 18 wurden bereits 1995 von der IARC als Karzinogene eingestuft. Dies gilt nach neuesten Erkenntnissen auch fur die die E6- und E7-Gene weiterer sog. Hochrisiko-HPV-Typen [28]. Eine persistierende Infektion mit Hochrisiko-HPV ist notwendige Voraussetzung fur die Entwicklung einer zervikalen intraepithelialen Neoplasie (CIN) III [4, 15]. Jedoch sind Kofaktoren fur dieses Geschehen unerlasslich. Diese sind noch unzureichend bekannt. Sicher ist jedoch, dass einer Immunsuppression, dem Rauchen und weiteren genitalen Infektionen (am besten definiert fur Chlamydien) Bedeutung zukommt. Mittlerweile gilt der Wert einer HPV-Testung sowohl im Screening als auch in der Beurteilung von Patientinnen mit auffalligen zytologischen Abstrichen als gesichert [15].

Neoplasias associated with anogenital human papilloma viruses (HPV) are characterised by high patient morbidity and mortality and by appreciable limitations in the patientʼs quality of life. Each year 530,000 women worldwide and 4800 women in Germany develop cervical cancer 1, 2. Biomolecular and epidemiological studies carried out in the past 25 years have demonstrated causal link between persisting infections with HPV 16 and HPV 18 and at least 11 other so-called high-risk HPVs (HR-HPVs) and the development of cervical cancer and its precursor lesions (so-called dysplasias or, respectively, cervical intraepithelial neoplasias – CIN). HPV 16, HPV 18 and other HR-HPVs are also the causes of other cancers and their precursors, for example, vulvar, vaginal, penile and anal cancers as well as tonsillar and throat cancers and certain forms of skin cancer. So-called low-risk HPVs (LR-HPVs) such as HPV 6 and HPV 11 are responsible for over 90 % of anogenital condylomata acuminata (anogenital warts). Condylomata acuminata are the most common viral sexually transmitted disease (STD) worldwide 3. It is estimated that around 1 % of European and German populations (aged 15–49 years) have these benign but often very unpleasant tumours. The development of a prophylactic quadrivalent vaccine (HPV 6, 11, 16, 18) and a bivalent vaccine (HPV 16, 18) has made it possible to prevent infections of the cervical epithelium and other squamous epithelia and the development of precancerous lesions. In the case of the quadrivalent vaccine (HPV 6, 11, 16, 18), the development of condylomata acuminata can also be prevented. The Standing Committee on Immunisation of the Robert Koch Institute (STIKO) has published a recommendation on HPV vaccination. Based on data from studies on the efficacy of HPV vaccines for the prevention of precancerous lesions of the cervix, vagina, and vulva, the STIKO recommends immunisation for girls aged between 12 and 17 years. The current guidelines do not contradict this recommendation but rather provide a more comprehensive supplement. The S3 guidelines focus on prophylactic vaccination against HPV-16 and HPV-18 or, respectively, HPV-6 and HPV-11 infections and thus on the prevention of cervical, vulvar, vaginal, anal and penile cancer and their precursors as well as on the primary prevention of condylomata acuminata and laryngeal papillomas. This S3 guideline thus clearly differs from other guidelines such as the S1 guideline “Condylomata acuminata and other HPV-associated clinical entities of the genitals, anus and urethra” (Guideline of the German STI Society in cooperation with the German Dermatological Society and the Paul-Ehrlich Society) and the S2 guideline of the German Society for Gynaecology and Obstetrics “Prevention, diagnostics and therapy for HPV infections and HPV-associated pre-invasive lesions in gynaecology and obstetrics”. After the debate on HPV immunisation has been concluded, the S3 guideline for the prevention of cervical cancer issued by the German Society for Gynaecology and Obstetrics will follow the recommendations in the current guideline, which will be updated. The development process and most important contents of the guideline are briefly described below. The long version of the guideline gives a more detailed account (www.awmf.org).

The case of a 22-year-old man suffering from genital warts is described. The lesions responded completely to recombinant interferon alfa-2a only after discontinuation of cannabis consumption. Cannabis was detected using the enzyme immunoassay/1-trans-tetrahydrocannabinoid method in urine. Southern blotting of frozen genital wart biopsy material revealed papillomavirus type 11 DNA, the amount of which increased significantly during interferon treatment. The final clearing of lesions after discontinuation of cannabis consumption implicates that the drug-induced impairment of cellular immunity was reversible. It is concluded that drug abuse and especially cannabis consumption may play some role in the world-wide increase in genital papillomavirus disease and in the high number of recalcitrant courses of genital warts.

Cervical carcinoma and its precursors are discriminated by their epidemiology from other malignant diseases. Human papillomavirusus (HPV) have been identified as the central etiologic factor in these diseases. Certain HPV gene products are tumor specific antigens in cervical cancer which leads to new perspectives in prevention and therapy. HPV testing is a realistic option in screening, triage and follow-up of cervical cancer and its precursors which has shown significant advantages over traditional methods in numerous large studies and proven cost effectiveness in recent model simulations.

A differential polymerase chain reaction (PCR), which takes account of both the human papillomavirus (HPV) type and the amount of viral DNA, was applied to 100 cervical intraepithelial neoplasias (CIN) in order to improve the characterisation of the lesions. One hundred women with cytological smears suggestive of CIN I-III and a colposcopically guided punch biopsy were studied. DNA was phenol-extracted and a consensus PCR with L1 primers and a type-specific PCR for HPV-16 and-18 E6 were performed in parallel. The different sensitivity of the PCR methods allowed a semiquantitative analysis of viral DNA. HPV analysis was possible in all 100 very small cervical punch biopsies. Of the lesions 79% were HPVpositive. The percentage of HPV-16-positive lesions increased significantly (P0.001) with the degree of severity of CIN (CIN I: 46%, CIN II: 63%, CIN III: 85%). The percentage of CIN with a high viral load increased also with the grade of CIN (CIN I: 8%, CIN II: 33%, CIN III: 44%). Cytologically positive lesions (Pap IIID or IVa) had significantly more frequently (p0.02) a high viral load: 39% vs. 10% of cytologically false negative lesions. In conclusion, type-specific PCR had a very high sensitivity in the detection of HPV-DNA also in cytologically false negative CIN. The addition of the less sensitive consensusPCR allowed a semiquantitative analysis of the viral copy number. The higher amount of viral DNA in cytologically positive lesions, which may correspond to a higher rate of proliferation, reflects a possible role of the viral load in the progression of CIN. In clinical practice, differential HPVPCR could help to improve the management of CIN.

Das gleichzeitige Auftreten von Thrombozytopenie, Anstieg der Transaminasen und Hamolyse bei praeklamptischen Patientinnen wurde erstmals 1954 von Pritchard et al. [1] ausfuhrlich beschrieben. 1982 fuhrte Weinstein [2] den Begriff „HELLP“ fur diesen Symptomenkomplex ein.

A 27-year-old woman suffering from panmyelopathy for six years presented with a cervical low grade squamous intraepithelial lesion (SIL), vulvar high grade SIL and perianal squamous cell carcinoma with an inguinal metastasis. Southern blot hybridization with 32P-labeled human papillomavirus (HPV) DNA revealed HPV 16 DNA in varying copy numbers in material from the four locations. HPV 16 genomes persisting after surgery on the perianal tumor area were no longer detectable after betatron radiotherapy.

Between 1988 and 1991 thirty-nine pregnant women suffering from HELLP-Syndrome were treated in our hospital. A retrospective analysis of clinical course and of the changes in the relevant laboratory values was performed. The incidence of the HELLP-Syndrome at Freiburg University Hospital was 0.85% for all pregnancies or 17% of the patients with toxaemia. 90% of the women were primipara, there was no prevalence of a certain group. The clinical course was characterized by the perseverance of symptoms for more than two weeks in 18% of the cases and a perinatal mortality of 25%, the latter mainly due to prematurity of the infants. This suggests that perinatal mortality due to respiratory distress syndrome can be reduced by conservative and expectative management of these patients. However on the other side a high rate of caesarean sections (77%) and sometimes even induced abortions were necessary to avoid maternal mortality and severe pre-or postpartal complications for these women.

Former examinations showed that 60% of cervical intraepithelial neoplasias are associated with the papillomavirus types 6/11 and 16/18 [2]. Papillomaviruses, particulary type 16/18, are said to have an effect on the development of cervical carcinoma [3]. Thus, a mode of therapy for precursors of cervical carcinoma should also remove a possible carcinogen. The aim of our investigations was to evaluate the therapeutic success of laser treatment on the cervix, with particular regard to HPV findings and the degree of severity of the intraepithelial neoplasia being treated.

Seit etwa 15 Jahren werden humane Papillomviren (HPV) als mogliche Cofakto-ren bei der Entstehung des menschlichen Zervixkarzinoms diskutiert. In bis zu 90% der primaren invasiven Zervixkarzinome kann HPV-DNA verschiedener Typen nachgewiesen werden. Die Ergebnisse zahlreicher zell- und molekularbiologischer Untersuchungen aus den letzten Jahren deuten auf eine kausale Bedeutung dieser Viren bei der Karzinogenese an der Zervix hin (zur Hausen 1989).

The presence of human papillomavirus (HPV) DNA in invasive carcinomas of the vagina, in their lymph node metastases, and in corresponding normal tissue was investigated by Southern blot hybridization with 32P-labeled HPV DNA. Tumor tissue from ten of 18 patients with vaginal carcinoma contained HPV DNA. Three of the 18 patients had a history of cervical neoplasia more than 14 years before the diagnosis of vaginal carcinoma. Five of 15 primary squamous cell carcinomas, one primary adenocarcinoma, and a vulvar recurrence of a vaginal squamous carcinoma contained HPV 16. A primary squamous carcinoma yielded HPV-related sequences. The HPV copy number varied from 0.5 to 50 per cellular genome. Four histologically positive inguinal lymph nodes from three patients contained HPV DNA. In six tumor-free control tissues from four patients, no HPV DNA was detected. No relationship was established between HPV positivity, HPV type, or copy number of the tumor and the grade of differentiation or keratinization or the clinical stage. After a median follow-up of 13 months, five of nine HPV-positive patients were alive without recurrence, whereas all seven HPV-negative patients had died because of disease. The results of this study indicate a possible major role of HPV in the development of vaginal cancer.

In 147 patients with suspected dysplasia, the effect of HPV 16/18 infection on the spontaneous course of the cervical intraepithelial neoplasia (CIN) and on the success of conisation and laser treatment was studied. The patients with PAP II smears had a high incidence of HPV (67%). This was accompanied, however, by a high rate of regression and fluctuation of the various virus types. Of the untreated cases of CIN, the lesions associated with HPV 16/18, particularly after repeat virus DNA identification of this type, had a higher persistence rate than HPV 16/18 negative CIN. After local therapy by laser surgery or conisation of HPV 16/18-related CIN, persistent disease must be expected in 30%. CIN recurrence was significantly more frequent in HPV 16/18 positive cases. According to the results of the investigations, HPV typing appears to be a useful tool in deciding on the appropriate therapy only in cases of persistent CIN and in determining an HPV 16/18 positive at-risk group after local (laser/conisation) therapy.

Cancer of the cervix has the epidemiologic characteristics of a sexually transmitted disease (Rotkin 1973). During the past decade the human papillomaviruses (HPV) have emerged as the most probable venereal agent in the pathogenesis of this disease.

DNA of a new papillomavirus type was cloned from a cervical carcinoma biopsy. Two EcoRI clones of 7.8 and 6.9 kb in length were obtained, the latter contained a 900-bp deletion. The BamHI fragments of both clones were used to characterize the DNA. It represents a distinct type of papillomavirus as determined by its size, its cross-hybridization with DNA of other papillomavirus types under conditions of low stringency only, the co-linear alignment of its genome with HPV 6 and HPV 16 prototypes and its occasional occurrence as oligomeric episomes. We tentatively propose to designate it as HPV 18. DNA hybridizing with HPV 18 under stringent conditions was detected in 9/36 cervical carcinomas from Africa and Brazil, in 2/13 cervical tumors from Germany and 1/10 penile carcinomas. Benign tumors (17 cervical dysplasias, 29 genital warts), eight carcinomata in situ and 15 biopsies of normal cervical tissue were devoid of detectable HPV 18 DNA. HPV 18-related DNA was found, however, in cells of the HeLa, KB and C4-1 lines all derived from cervical cancer. The state of the viral DNA was investigated in four cervical cancer biopsies. The data reveal that the DNA might be integrated into the host cell genome. One tumor provided evidence for head to tail tandem repeats some of which persisted as circular episomes.

Using Southern Blot hybridization 18 invasive malignant tumours of the vulva and 2 intraepithelial vulvar neoplasias were tested for the prevalence of HPV-DNA. In 6 of 11 primary squamous carcinomas, including one carcinoma of the Bartholinic gland, HPV 16 could be identified. Contrarily, in all seven recurrent vulvar tumours examined, including 5 squamous carcinomas, 1 fibrosarcoma and 1 zylindroma, no HPV-DNA was found. No correlation existed between HPV status of the tumours and grade of differentiation or keratinization, or the tumour-stage and lymph-node status. In one tumour-invaded lymph-node HPV 16 was detected as in the corresponding primary tumour. 12 samples from tumour free abdominal skin considered as corresponding normal tissue specimens were negative for HPV-DNA. A correlation between clinical course of the disease and HPV status of the primary tumours was not observed.

Demonstration of human papilloma virus DNA (HPV-DNA) in the lesions of focal epithelial hyperplasia in an 11-year-old Turkish girl provides further proof of its viral cause. In this case characterization of HPV-DNA showed a reaction with that of type 13 and 18. This previously undescribed variant is probably the expression of a new HPV type.

The development of bowenoid papules in a 20-year-old man and a carcinoma in situ of the portio uteri of the 22-year-old female sexual partner is reported. In both lesions HPV-16 DNA could be detected by molecular biological means. This observation led us to the conclusion that HPV 16 had been transmitted sexually. The same seems to be true for HPV-11-induced condylomata acuminata, which appeared on the external genitalia of both patients after recuperation from the HPV-16-induced lesions and conisation treatment. The clinical significance of this observation and its consequences for dermatologists and gynecologists are discussed.

• In biopsy specimens from a patient with tracheal and bronchial papillomatosis, human papillomavirus (HPV) type 11 DNA was identified. Treatment with leukocyte interferon was initiated, and the results of therapy was monitored by molecular hybridization of biopsy specimens with phosphorus 32-labeled HPV type 11 DNA after interferon application. There was no improvement of the clinical course, although two of five biopsy specimens showed a considerable reduction in the amount of HPV type 11. We discuss the reasons for the insufficient effect of therapy and propose molecular hybridization as an additional method for therapy control in viral diseases. ( Arch Otolaryngol 1985;111:96-98)

In one out of 22 cases of histologically diagnosed bowenoid papulosis, we could detect human papilloma virus (HPV) capsid antigen in paraffin sections fixed with formalin by means of the peroxidase-antiperoxidase (PAP) test; there were no positive findings regarding biopsies taken from Bowen's disease. Southern blot hybridization technique revealed HPV 16 DNA in 8 out of 10 cases of bowenoid papulosis; there was one positive identification out of 5 cases of Bowen's disease.

A series of 47 lesions diagnosed cytologically as cervical intraepithelial neoplasia (CIN) III in 22 cases, CIN I/II in 13 cases, and 12 cases showing abnormal smears consistent with human papillomavirus infection were analyzed. Thirty-six cases with negative cytology were used as the control group. Sixty-eight percent of CIN III were positive for a mixture of human papillomavirus 16 and human papillomavirus 18, 18% reacted with human papillomavirus 6 or 11, and 14% were negative. Of the group with CIN I/II or with abnormal Papanicolaou smears, approximately one-third contained human papillomavirus 6 (11) and one-third human papillomavirus 16 and 18. Only 11% of the samples from the control group hybridized with human papillomavirus 6 (11), the others were negative with either probe. The data obtained by the rapid in situ hybridization of cervical cells are in agreement with the presence of human papillomavirus 16 and 18 in a high proportion of cervical carcinoma and carcinoma in situ lesions. Thus, the method can be applied to test the hypothesis that a lesion containing human papillomavirus 16/18 positive cells has a higher risk of progressing to cancer than a lesion harboring human papillomavirus 6 or 11.

We report on a 17-year-old patient suffering from generalized verrucosis. This diagnose had been confirmed by means of light microscopy, electron microscopy, and DNA hybridisation (HPV type 2). The study of immunological parameters revealed reduced numbers of T-lymphocytes, especially of helper-T-cells. Skin tests with recall antigens were normal. Three brothers of our patient showed slight reduction of helper-T-cells. Treatment with topical retinoic acid was not effective. Treatment with alpha-interferon i.m. intralesionally was effective in the beginning, but two weeks after onset of therapy no further improvement could be achieved. Discontinuation of therapy was followed by a relapse. Tigason which was given thereafter led to flattening of the warts but failed to bring about complete regression within the following four months.

Persistent condylomata acuminata in a 21-year-old patient with diabetes mellitus were treated with highly purified interferon-alpha (IFN-alpha) obtained by recombinant DNA technology. Daily dose was 1.5 X 10(6) IU, given subcutaneously. Already during treatment the condylomata regressed. Two weeks after the end of therapy, i.e. after a total dose of 10.5 X 10(6) IU IFN-alpha, all condylomata had completely receded. Blood glucose levels remained constant with concomitant insulin therapy. Toxic side-effects or antibodies to IFN-alpha were not observed.

In 104 woman patients with cervical intraepithelial neoplasia (CIN) I-III who had been treated between 1981 and May 1986 in the dysplasia consulting-room of the Department of Gynecology by means of conisation effected by laser, we checked on therapeutic success via cytology and colposcopy as well as by human papilloma virus (HPV) typing. In 90.3% of the cases 1.25 laser treatments sufficed for healing of the lesion. Relapses occurred mainly in severe intraepithelial neoplasias with identified HPV 16/18. Incidence of HPV was reduced from 56% before therapy to 13.3% after therapy. This shows that laser therapy can be regarded as an effective treatment of the causes of the disease even in CIN associated with HPV. The markedly increased relapse rate in CIN III (24%) makes a reliable follow-up mandatory especially in such cases.

Using Southern Blot hybridization 18 invasive malignant tumours of the vulva and 2 intraepithelial vulvar neoplasias were tested for the prevalence of HPV-DNA. In 6 of 11 primary squamous carcinomas, including one carcinoma of the Bartholinic gland, HPV 16 could be identified. Contrarily, in all seven recurrent vulvar tumours examined, including 5 squamous carcinomas, 1 fibrosarcoma and 1 zylindroma, no HPV-DNA was found. No correlation existed between HPV status of the tumours and grade of differentiation or keratinization, or the tumour-stage and lymph-node status. In one tumour-invaded lymph-node HPV 16 was detected as in the corresponding primary tumour. 12 samples from tumour free abdominal skin considered as corresponding normal tissue specimens were negative for HPV-DNA. A correlation between clinical course of the disease and HPV status of the primary tumours was not observed.

Human papillomavirus (HPV) standardization and DNA cytophotometry were carried out in 29 cases of cervical intraepithelial neoplasia (CIN) grades I-III. A prognostically unfavorable DNA distribution pattern with an aneuploid stemline was found in 14 of the 16 dysplasias with HPV types 16 and 18, while in 11 of 13 dysplasias with HPV types 6 and 11 there was a favorable DNA distribution with a euploid-polyploid stemline. Among 178 colposcopically, cytologically and histologically confirmed cervical lesions, there was a statistically significant incidence of HPV 16/18 infections in severe dysplasias and carcinomas, while HPV 6/11 was found predominantly in mild cervical lesions. It seems that CIN can be divided into high- and low-risk lesions not only by the degree of severity and the DNA distribution pattern but also by HPV typing.

Immunohistochemical staining of epidermal growth factor receptor (EGF-R) with a monoclonal antibody was performed in 43 biopsies of cervical tissue. The distribution of the receptors in normal cervical tissue differs from that of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma of the cervix. Whereas the staining reaction in normal squamous epithelium was confined to the basal and deep parabasal cell layer, in all cervical intraepithelial neoplasias, with or without human papilloma virus association, a homogeneous EGF-R staining reaction could be observed throughout the entire lesion. This means that the dysplasia cells of a CIN I-III, like the tumor cells of a squamous cell carcinoma, have a raised EGF-R content, which in the normal squamous epithelium is usually only found in the basal and deep parabasal cells that are capable of dividing. No EGF-R staining reaction could be detected in the higher, differentiated cell layers of the normal squamous epithelium of the cervix.

Papillomavirus infections in the genitoanal region are detectable by the demonstration of type-specific HPVDNA (DNA-DNA hybridization, in situ hybridization on cell smears) or by a peroxidase-antiperoxidase assay using formalin-fixed paraffin sections for demonstration of common structural antigens of papillomaviruses. Using these methods an epidemiological study on sexual partners with genitoanal papillomavirus infections has been initiated.