尖锐湿疣患者疣体HPV多型别原位杂交检测

目的　探讨原位分子杂交技术多型别检测尖锐湿疣患者疣体HPV DNA的敏感性。方法　应用原位杂交法 ,通过广谱生物素标记的HPV DNA探针 ,对尖锐湿疣患者疣体HPV 6,11,16,18,3 0 ,3 1,3 3 ,3 5 ,45 ,5 1,5 2型DNA进行检测。结果　在 3 5例标本中 ,有 3 3例HPV DNA呈阳性反应 ,阳性率 94.3 %。阳性反应物主要分布于表皮浅层空泡细胞的细胞核内 ,多呈片状或灶状分布。结论　用广谱生物素标记的原位杂交技术 ,可检测HPV多型别 ,具有敏感性高、特异强的优点 ,并能进行病毒的组织学定位     

Nestin expression in Bowen's disease and Bowen's carcinoma associated with human papillomavirus

Human papillomaviruses (HPVs) have been detected in lesions of Bowen's disease (BD) and Bowen's carcinoma (BC); the invasive tumor retains the cytological characteristics of BD. Previous reports suggest that nestin-expressing hair follicle stem cells are undifferentiated and pluripotent, and nestin expression in some tumors indicates poor differentiation and high grade of malignancy. We identified HPV-DNA in BD (n=25) and BC (n=23) by in situ hybridization (ISH) analysis with INFORM(?) HPV III (Ventana Medical Systems. AZ, USA) and determined nestin expression by indirect immunohistochemical staining with anti-nestin polyclonal antibody (IBL, Gunma, Japan). We detected HPV-DNA in 68% of BD and in 87% of BC. In BD, 13 cases demonstrated the punctuate pattern, and four showed nestin expression. In BC, 19 cases showed the punctuate pattern and 16 showed nestin expression. HPV-DNA integrates into the host genome, and this is observed as the punctuate pattern on ISH. The nestin expression was statistically high in group of BC than BD (P<0.01). These results therefore suggest that HPV-DNA integrated in the genome of tumor cells of these diseases and contributed to malignant alteration. From the standpoint of tumorigenesis, BC might represent one type of poorly differentiated, high-grade squamous cell carcinoma.     

Penile cancer among patients with genital lichen sclerosus

BACKGROUND: Genital lichen sclerosus (LS) has sporadically been reported to be associated with penile squamous cell carcinoma (SCC). OBJECTIVE: The purpose of this study was to assess the risk of malignant degeneration in a series of male patients affected by genital LS. METHODS: All cases of histologically proven epithelial malignancy associated with penile LS recorded in our pathology files over a 10-year period (1987-1997) were reviewed. Assessment for presence of human papillomavirus (HPV) was performed from paraffin-embedded tissues using polymerase chain reaction (PCR). RESULTS: Five of 86 white and uncircumcised men with genital LS (mean age at diagnosis, 53 years; range, 22-83 years) showed malignant or premalignant histopathologic features: 3 had SCC, one had erythroplasia of Queyrat (unifocal SCC in situ), and one verrucous carcinoma. The average lag time from onset of LS was 17 years (range, 10-23 years). Histologically, transition from LS to frank neoplastic foci was evident in all cases of SCC. In these SCC cases, areas of epithelial dysplasia were well evident at the tumor periphery. In the remaining cases, the histologic findings were consistent with erythroplasia of Queyrat and verrucous carcinoma. PCR detected HPV 16 infection in 4 of the 5 cases; one SCC patient was negative for HPV. CONCLUSION: Malignant changes were associated with 5.8% of the cases of penile LS in our series. Therefore patients with genital LS are at considerable risk of the development of penile SCC, as well as other epithelial and in situ carcinomas, namely verrucous carcinoma and erythroplasia of Queyrat. HPV infection probably plays a major role because 4 of 5 patients were positive for HPV. Histologically, epithelial dysplasia may represent a precancerous stage before the development of neoplasia in atrophic nonproliferative LS lesions, as its presence at the tumor periphery in our SCC biopsy samples seemed to suggest.     

Incidence of preputial lichen sclerosus in adults: histologic study of circumcision specimens

BACKGROUND: Narrowing of the prepuce in men is poorly documented, and the causes are often unknown, except in the case of clinical infections or skin diseases such as lichen sclerosus (LS). OBJECTIVE: We conducted a histologic study of circumcision specimens with phimosis or paraphimosis. METHODS: This prospective study included 43 men with contraction referred for circumcision. RESULTS: LS was present in 32% of cases, but only 12% of these cases of LS had not been diagnosed before circumcision. In 31% of cases the histologic findings were normal. Subacute nonspecific inflammatory changes were diagnosed in 37% of all cases, and secondary narrowing of the prepuce in 62% of cases. It is probable that this histologic modification of the preputial mucosa is involved in narrowing of the prepuce. CONCLUSION: Phimosis in young adults is usually not associated with LS (only 14%). In contrast, most older patients had secondary phimosis caused by progressive LS (40%) or subacute nonspecific inflammatory changes (40%). Although all cases of phimosis in men should be treated by complete circumcision to prevent penile cancer, paraphimosis associated with preputial dyspareunia, with the exception of cases associated with LS, can be treated by corrective surgery.     

Lichen sclerosus is frequently present in penile squamous cell carcinomas but is not always associated with oncogenic human papillomavirus

BACKGROUND: Penile squamous cell carcinoma (SCC) may occur on pre-existing lesions of lichen sclerosus (LS). However, the prevalence of histological changes of LS in penile SCC is not well established. Moreover, mucosal oncogenic human papillomaviruses (HPVs) are sometimes detected in penile SCC, but have not been systematically sought in LS-associated penile SCC. OBJECTIVES: To establish the prevalence of LS histological changes and of mucosal oncogenic HPV in a series of patients with penile SCC. METHODS: Consecutive cases of histologically proven penile SCC from a single university hospital over a 14-year period were retrospectively selected and reviewed. Histological signs of LS were systematically sought. HPV was detected by polymerase chain reaction (PCR) amplification of DNA from paraffin-embedded skin samples using general primers GP5+/GP6+ (allowing detection of mucosal HPV) and oncogenic type 16-, 18-, 31- and 33-specific primers. RESULTS: Eighteen cases of penile SCC were found. The mean +/- SD age of patients at diagnosis was 67.3 (14.5 years). In eight of 18 (44%) cases, SCC was associated with histological features of LS. Seventeen skin biopsy specimens of SCC (nine without and eight with LS histology) were subjected to PCR amplification for HPV. Mucosal HPV was detected in six of them (35%). Five of nine SCCs without histological features of LS were positive for mucosal HPV: three with HPV type 16 and two with only general primers. In contrast, all eight SCCs associated with LS were negative for oncogenic HPV types, although one was positive with general primers. CONCLUSIONS: Penile SCC seems to be frequently associated with LS histological changes. As with vulval SCC, we found that non-LS-associated penile SCC tended to be frequently associated with oncogenic HPV infection, whereas LS-associated penile SCC was not. Larger series are needed to confirm this association.     

Oncogenic mucosal human papillomaviruses in Bowen's disease of the hands

INTRODUCTION: The association between genital Bowen Disease (BD) and human papillomavirus (HPV) especially HPV-16 infection is well known, but it is more rarely related to extragenital BD. The aim of this study was to detect the presence of oncogenic HPV infection in the BD of the hands considering the histology and the presence of oncogenic HPV detected by in situ hybridization (ISH) and polymerase chain reaction (PCR). MATERIAL AND METHODS: Eleven formalin-fixed and paraffin-embedded samples of BD of the hands were selected. We looked for koilocytosis using standard histological procedure. ISH was performed using genomic HPV DNA probes types 16, 18 and 33 labeled with digoxigenin. Next, with a new amplification system using biotinyled tyramide (Kit Dako Gen Point K620) was used to detect hybrids. Otherwise, Baay's HPV type specific primers (type 16, 18, 31 et 33) were chosen for PCR. RESULTS: Koilocytosis were observed in all cutaneous samples. HPV 16 was detected in 9/11 cases (82 p. 100): 2/10 with ISH and 9/11 with PCR. DISCUSSION: We report here the largest series of BD of the hands, associated with HPV type 16 infection. This high rate (82 p. 100) compared to the other series is linked to the high sensitivity of PCR and is increased by the choice of Baay's oligonucleotide primers. Besides, the low HPV rate (2/10) by ISH, is similar to those obtained in the other series. Although ISH is less sensitive than PCR, the morphological study was useful to argue the pathogenicity of HPV. Finally, the high prevalence of genital oncogenic HPV on the hands, plaid in the scratching hypothesis resulting in autoinoculation from HPV lesions in the genital region.     

尖锐湿疣人乳头瘤病毒颗粒、核酸及抗原检测

42例尖锐湿疣进行了HPV-DNA序列,HPV抗原和电镜检查。36例(85.7%)HPV-DNA序列检测阳性。其中单一型感染9例,混合型感染27例。以HPV_(11)阳性最多,计32例,HPV_(6b)26例,HPV_(16)19例,HPV_(18)6例。HPV抗原阳性28例(66.7%)。15例查见病毒颗拉(35.7%)。总计阳性率为90.5%(38/42例)。文中对三项检查结果作了比较和分析。     

原位杂交和免疫组化技术检测尖锐湿疣的比较

目的比较原位杂交和免疫组化检测HPV的优劣,从而找到一种较确切的病理诊断尖锐湿疣的方法。方法对29例门诊诊断为尖锐湿疣(CA)的蜡块作连续切片,分别作原位杂交(ISH)和免疫组化(IHC)对比染色,在光镜下观察。结果原位杂交检测HPV6B/11阳性25例(86.20%);免疫组化检测HPV6/11阳性16例(55.10%)。两者差异有统计学意义(P<0.01)。在连续切片上,原位杂交检测HPV阳性细胞比免疫组化检测的明显增多而且有效的降低了背景染色。结论 HPV原位杂交技术与免疫组化相比,其特异性好,灵敏度高,为尖锐湿疣的确诊及临床治疗提供更准确、可靠的依据。     

Lymphohistiocytic and granulomatous phlebitis in penile lichen sclerosus

Lichen sclerosus (LS) is a chronic inflammatory disease of unknown etiology that may affect the genital and/or extragenital skin of individuals of either sex at all ages. In boys, the prepuce is the most common site of involvement. The diagnostic criteria of LS include the presence of inflammatory infiltrates mainly composed of T lymphocytes. We report on two cases of LS of the prepuce because of the unusual feature of lymphocytic (CD45RO+ and CD20+), histiocytic (CD68+), and granulomatous phlebitis. This lesion was not present in a group of another 18 cases of childhood penile LS. We have not been able to find any references describing and illustrating inflammatory involvement of the dermal vein walls in LS. Unlike the data reported in the literature, the dermal inflammatory infiltrates of these two cases showed a similar proportion of B and T lymphocytes in addition to frequent CD68+ histiocytes.     

尖锐湿疣组织HPV基因芯片分型及型别分析

目的研究尖锐湿疣(CA)组织中人乳头瘤病毒(HPV)感染的病毒亚型分布,CA患者中高危型HPV感染的情况。方法采用基因芯片检测和分型方法,对武汉市302例CA患者的皮损组织进行HPV检测及分型。结果302份CA组织标本中HPV阳性286例,检出率为94.7%,单一型别阳性率为73.8%,多重型别阳性率为20.7%,低危型别(HPV6,11,42,43,44)阳性率为80.5%,高危型别(HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83,MM4)阳性率为14.2%,对照组阳性率为16.7%。结论本地区尖锐湿疣以单一型别及低危型别HPV感染为主,高危型HPV感染阳性率为14.2%。     

Genital carriage of human papilloma virus (HPV) DNA in prepubertal girls with and without vulval disease

Human papilloma virus (HPV) can reach a child's anogenital area by vertical transmission or by close contact, which can be either sexual or nonsexual. Our objective was to compare HPV in prepubertal girls with and without lichen sclerosus (LS). We compared the frequencies and types of HPV in girls with LS with those in children with non-LS vulval disease (vulval swab and urine) and in children with no known vulval disease (urine only). HPV DNA was detected using a nested polymerase chain reaction (PCR) with general and consensus primers amplifying a region of the L1 gene, and PCR amplicons were typed using reverse hybridization with labeled HPV type-specific probes. Specimens untypeable by this method were typed by DNA sequencing. In the cohort of children with LS, we recorded the presence of maternal anogenital warts or a dysplastic cervical smear within 3 years of the affected child's birth. We found that HPV was present in the urine and vulval swabs of 8 of 32 children with LS and in 2 of 31 children with non-LS vulval disease, but also in the urine of 7 of 29 controls. In those with LS, the frequency was not increased significantly, but the types were predominantly those commonly associated with dysplasia of the cervix, penis, vulva, and anus, as opposed to the broader spectrum of types found in the control group, not all dysplasia associated. Two of the 32 mothers reported warts, and 15 of 32 (46.9%) had an abnormal smear. (The national average of abnormal cervical smears is less than 10%.) We concluded that HPV appears to be common in all prepubertal girls, but children with LS carried types associated with dysplasia and their mothers had had a high incidence of dyskaryotic smears.     

Human papillomavirus in cervix carcinoma and condylomata acuminate—identification of HPV-DNA by improved dot-blot-hybridization

Using the benzoylated naphthoylated DEAE cellulose method (BND-method) we have designed a more efficient approach for the detection of human papillomavirus-DNA (HPV-DNA) via dot-blot and hybridization. Biopsy material from anogenital warts (40 patients), invasive carcinoma uteri (12 patients) and normal controls (20 patients) were studied for the presence of HPV-DNA. Phenol extracted DNA from representative lesions was loaded onto a pretreated nitrocellulose filter, was incubated under stringent conditions with 32-P-dCTP labelled HPV-DNA and exposed to a Kodak X-OMAT film. DNA of HPV types 11, 16, 18 were cloned into plasmid vectors. The common, time-consuming caesium-chloride density-gradient centrifugation used for purification of plasmid DNA (20-36 h), was substituted by the BND-method (15 min). Complete HPV genomes were excised using the restriction endonucleases Eco RI and Bam HI. The HPV-DNA fragments obtained were then electroeluted using the 'Bio-Trap' method and subsequently labelled with 32P-dCTP by nick translation. Without resorting to more complex and sensitive technology, such as the polymerase chain reaction, efficiency of specific analysis of large numbers of cervical samples and condylomata was achieved without loss of accuracy or increased costs. The time required for HPV identification from biopsy or sample receipt was shortened considerably (approximately 50%).     

Diagnosis of genital human papillomavirus (HPV) lesions in the male: correlation of peniscopy, histology and in situ hybridisation.

OBJECTIVE--To assess the diagnostic criteria of genital HPV lesions in male sexual partners of HPV infected women. METHODS--Peniscopically directed biopsy specimens (from 693 lesions in 300 men) were examined on light microscopy and in situ hybridisation (ISH) for HPV types 6,11,16,18,31,33 and 42. The predictive value of different histological criteria for ISH positivity was also evaluated using stepwise logistic regression analysis. RESULTS--Flat HPV lesions were most accurately predicted by the punctuation pattern on peniscopy, giving the concordance between peniscopy and histology between peniscopy and histology of 79.5% (66/83) and that between peniscopy and ISH of 56.6% (47/83). Diffuse acetowhite pattern disclosed a typical HPV lesion in only 17.8% (13/73), and HPV DNA was found in 11.0% (8/73) of cases. Of the 114 biopsy specimens from peniscopically healthy areas adjacent (0.5-1 cm) to the lesions, 93.0% (106/114) were normal on light microscopy, and HPV DNA was found in only 2.6%. Penile intraepithelial neoplasia (PIN) lesions were most frequently ISH positive, 81.1% (30/37), 50% showing HPV 16 and/or 18 DNA. Lesions classified as HPV-suspicious or nonspecific on light microscopy were HPV DNA-positive in 16.9% (11/65) and 8.1% (13/160), the frequency of high-risk HPV types being 3.1% and 1.3%, respectively. In logistic regression analysis, koilocytosis was the most powerful predictor of ISH-positivity in the flat lesions (without PIN), the risk ratio being 3.7. CONCLUSION--No conclusive peniscopic criteria for male HPV infections could be established, making histological evaluation mandatory. Care should be exercised in interpreting as HPV lesions the cases devoid of koilocytosis, HPV typing being essential in confirming the diagnosis in doubtful cases.     

Papillomavirus-associated balanoposthitis.

OBJECTIVE--To assess whether there might be an association between genital papillomavirus infection (GPVI) and balanoposthitis. DESIGN--Retrospective HPV DNA examination of biopsy specimens from 23 men suffering from balanoposthitis and exhibiting acetowhite lesions that were penoscopically and histologically concurrent with HPV infection. SETTING--The STD clinics at Karolinska Hospital and South Hospital, Stockholm, Sweden. PARTICIPANTS--Randomly selected men attending with long-lasting and/or recurrent penile symptoms and exhibiting a clinical picture of balanoposthitis, who revealed a penoscopical and histopathological picture of epidermal lesions that were concordant with accepted criteria for typical or conspicuous GPVI. Asymptomatic controls were selected retrospectively on the basis of identical penoscopy and histology criteria. RESULTS--A history of previous condylomata was obtained in eight (35%) of 23 men. At penoscopic evaluation tiny condylomatous lesions were observed in five (22%) patients. The in situ hybridisation (ISH) assay using specific probes for the HPV types 6/11, 16/18, 31/33 and 42 was positive in 13/23 (56%) of the patient samples, but in only 26% of the 19 control samples. In patient biopsies the oncogenic HPV types 16/18 and/or 31/33 were found in 7/13 samples, whereas HPV 6/11 and/or 42 were present in another six cases. PCR performed on the ten ISH negative patient biopsies, were negative in all cases. CONCLUSION--Symptoms included redness, itching, burning, tenderness, dyspareunia, fissuring and in two cases penile oedema and inguinal adenopathy. All patients fulfilled penoscopical and histopathological criteria for HPV infection. We demonstrate some tentative evidence that HPV might be associated with long-lasting balanoposthitis, although our data still are circumstantial for a causative association. The results also elucidate the diversity in clinical presentation of GPVI.     

Known HPV types have no association with keratoacanthomas

Certain HPV types have been linked to the genesis and development of premalignent and malignant skin diseases. There have been several contradictory reports on the role of HPV infections in the development of keratoacanthomas (KAs). To further study the involvement of HPVs in the aetiology of KAs, we investigated paraffin-embedded specimens of 80 biopsies of KAs for the presence of HPV 1, 2, 3, 4, 5, 7, 26, 37, 38, 47 and 59 DNA by in situ hybridization (ISH) with biotinylated probes under high stringency conditions (Tm-10°C). Every fourth biopsy specimens was also examined by polymerase chain reaction (PCR) with consensus primers targeting the HPV E1 and L1 regions. The positive cases were further studied by direct DNA sequencing. All specimens proved to be negative for all HPV DNAs studied by ISH. Three out of 20 cases produced in positive PCR amplifications when consensus primers targeting the L1 region were used. However, the same samples remained negative with general primers targeting the E1 region. The DNA sequence analysis of the PCR-positive products showed a 76% homology with HPV type 17. Our results suggest that the known HPV types are unlikely to have any role in the aetiology of KAs.     

Simultaneously detected aberrant p53 tumor suppressor protein and HPV-DNA localize mostly in separate keratinocytes in anogenital and common warts

Abstract The E6 oncoprotein of human papillomavirus (HPV) is known to inactivate the control function on cell cycle exerted by p53 tumor suppressor protein in vitro by binding to p53 and thus facilitating the degradation of p53. We have applied a simultaneous in situ demonstration method for detecting p53 protein and HPV-DNA on formalin-fixed tissue sections, and investigated the in vivo interrelationship of p53 protein and HPV-DNA. Immunohistochemical staining for p53 protein with polyclonal and monoclonal antibodies, recognizing both wild-type (wt) and mutated p53 protein, was performed first and in situ DNA hybridization (ISH) for HPV types 6/11 or 16/18 with digoxigenin-labelled probes thereafter. 47% (25/53) of 48 histologically confirmed primary or recurrent condylomata acuminata (CA), 2 Bowenoid papulosis (BP) and 3 common wart (CW) biopsies, positive for HPV 6/11 or HPV 16/18 DNA, showed keratinocytes immunopositive for p53 protein. Of these. 11 lesions with abundant numbers of p53-positive cells were further analyzed with the double method. Signals for abnormal p53 protein and HPV-DNA were detected in separate cell nuclei in all biopsies and, additionally, in the same cell nuclei in 3 biopsies (1 BP, 1 CA, 1 CW). Usually the p53 positivity localized more basally in the epidermis than HPV-DNA, although p53- and HPV-positive keratinocytes were always located closely. The findings were similar for HPV-types 6/11 and 16/18. Our finding of both p53 and HPV-6/11 signals in the same cell nuclei may indicate complexing of p53 and low-risk HPV's without degradation of p53. Our results show abnormal p53 expression in HPV-infected skin lesions, and suggest that p53 protein is susceptible to aberrations even in the cells in the vicinity of productive HPV infection. However, it is not yet fully understood how HPV interferes with p53 protein in these cells.     

Association of penile lichen sclerosus and oncogenic human papillomavirus infection

BACKGROUND: Data on the prevalence of human papillomavirus (HPV) infection in patients with penile lichen sclerosus (LS) are scant and controversial. AIM: To investigate the prevalence of HPV infections in patients with penile LS. METHODS: HPV infection was assessed by polymerase chain reaction (PCR) in paraffin-embedded penile biopsies obtained from the glans or inner foreskin of 46 adult patients with penile LS, and in brush cytology smears of penile healthy mucosa from an equal number of randomly selected control males matched for age. Statistical evaluation was performed using conditional logistic regression analysis. RESULTS: PCR disclosed the presence of HPV infection in 17.4% of LS patients (HPV 16, six cases; HPV 18, one case; HPV 45, one case). Amongst the controls, HPV infection occurred in 8.7% of patients (HPV 16, two cases; HPV 53, one case; HPV 70, one case). Statistical regression analysis confirmed that the rate of HPV infection was higher amongst patients with genital LS than amongst healthy controls [odds ratio (OR), 2.55; 95% confidence interval (CI), 0.73-8.89]. CONCLUSIONS: Infection with oncogenic "high-risk" HPV types in patients with genital LS may enhance the risk of penile cancer arising on LS.     

Expression of PCNA is associated with the presence of HPV DNA in skin warts

A series of 90 excised cutaneous warts (verrucae vulgaris) were studied for the presence of HPV (human papillomavirus) DNA using in situ hybridization (ISH) with biotinylated full genomic DNA probes of HPV types 1, 2, 3 and 4. The expression of PCNA (proliferating cell nuclear antigen) was examined using conventional immunohistochemistry. The aim was to test the hypothesis that HPV can reactivate PCNA, including in the host replication machinery. HPV DNA of the above types was detected in 60 of 90 verruca biopsies studied (66.7%): HPV 2 in 56 cases, HPV 1 in 2 cases, and HPV 3 in 2 cases. PCNA was expressed in all samples except two. The signal distribution of HPV DNA markedly differed from that of PCNA expression. ISH revealed strong HPV DNA signals in both the granular and the upper spinous cell layers, the most intense signals being detected in the upper epidermis. On the other hand, nuclear PCNA staining was present in the majority of parabasal and basal cells. Although strong PCNA signals within the wart lesions were found in the areas where HPV DNA was present, the PCNA positivity was almost invariably localized in the differentiated cells of the spinous cell layers, just below the HPV DNA-expressing cells. At the margins of the lesions, PCNA expression was still strong but disappeared abruptly towards the normal epidermis. HPV DNA-positive warts showed more intense expression of PCNA than did the HPV DNA-negative ones in this study. Our results indicate that PCNA induction is associated with the presence of HPV DNA, suggesting that HPV can reactivate PCNA, thus interfering with the host cell DNA replication machinery. Received: 8 May 1996     

Cervical neoplasia and human papilloma virus infection in prostitutes.

OBJECTIVES--To evaluate the prevalence and incidence of PAP smears indicating cervical dysplasia as well as human papillomavirus (HPV) infection in prostitutes. DESIGN--Prevalence and incidence study of cervical dysplasia and HPV infection in prostitutes. For detection and typing of HPV-DNA In Situ Hybridisation (ISH) was performed in tissue samples with CIN gained by colposcopically directed punch biopsies. SETTING--Second Department of Obstetrics and Gynecology, University of Vienna Medical School and STD Clinic of the Public Health Office, Vienna. SUBJECTS--Registered prostitutes attending the STD Clinic of the Public Health Office and a control group. RESULTS--978 prostitutes and 5493 women with unknown cytological anamnesis were compared. Frequency of positive PAP smears was significantly higher in prostitutes (6.13% versus 1.43%). To determine the pick-up rate of cervical dysplasia during one year after negative cytology we compared 722 prostitutes and 3162 controls. Prostitutes showed a significant higher dysplasia pick-up rate (3.05% to 1.07%) compared with controls. HPV detection rate in prostitutes was similar to that in the control group. The distribution of HPV types revealed a higher frequency of "high risk" HPV 16/18 and 31/33 in prostitutes. CONCLUSION--The results demonstrate a higher incidence and prevalence of cervical dysplasia in prostitutes and therefore suggest regular cervical PAP smear screening in registered prostitutes twice a year.     

Assessment of the presence of mucosal human papillomaviruses in malignant melanomas using combined fluorescent in situ hybridization and chemiluminescent immunohistochemistry

BACKGROUND: The vast majority of studies aimed at detecting human papillomavirus (HPV) DNA in skin cancer have used sensitive polymerase chain reaction (PCR) methods but the PCR technique, despite its high sensitivity, is not suitable to ascertain whether (i) the presence of HPV can be related only to few cells harbouring the virus, (ii) the presence of HPV is due to a tumour surface contamination and (iii) the presence of HPV is localized in cancer cells, rather than in normal keratinocytes present in the tumour biopsy. In a recent work we have found mucosal high-risk (HR) HPV genotypes in primary melanoma by PCR. OBJECTIVES: To localize mucosal HR-HPV nucleic acids and tumoural melanocytic marker in the same sections of primary melanoma samples in order to understand the relationship between HPVs and melanoma cells. METHODS: We have developed a very sensitive method that combines an enzyme-amplified fluorescent in situ hybridization (ISH) for the detection of HPV nucleic acids (types 16 and 18) with a chemiluminescent immunohistochemistry (IHC) method for the detection of the tumoural melanocytic marker HMB-45 sequentially in the same section. Digital images of fluorescent ISH and chemiluminescent IHC were separately recorded, assigned different colours and merged using specific software for image analysis. RESULTS: The combined fluorescent ISH and chemiluminescent IHC demonstrated a sharp colocalization (in the range 60-80%) of HPV nucleic acids and melanoma marker inside the same sections of melanoma biopsies, with a strong specificity and sensitivity. CONCLUSIONS: The strong colocalization of mucosal HR-HPV nucleic acids and HMB-45 melanocytic marker emphasized that viral nucleic acids were specifically present in melanoma cells and supported a possible active role of HPV in malignant melanoma.