人乳头瘤病毒与皮肤Bowen病的相关性

目的 探讨人乳头瘤病毒(HPV)与皮肤Bowen病发病的相关性.方法 41例皮肤Bowen病患者皮损以及48例健康对照皮肤,采用多对引物,应用巢式PCR进行HPV DNA检测,同时应用半定量PCR进行病毒定量,对HPV DNA阳性标本进一步采用原位杂交方法分析组织内病毒的分布状况.结果 41例皮肤Bowen病患者皮损HPV DNA阳性检出率为12%(5例),其中黏膜型3例(2例HPV16,1例HPV33),病毒定量相当于101～103拷贝,原位杂交显示在多数肿瘤细胞核中有广泛阳性表达,而肿瘤邻近的正常组织无信号表达;皮肤型2例(HPV27和HPV76各1例),原位杂交均无阳性表达.此外,对照组中有1例检出HPV DNA,属于疣状表皮发育不良相关型HPV23,检出率为2.1%,与皮肤Bowen病皮损中HPV检出率比较,差异无统计学意义.2例患者皮肤型病毒定量较低,与正常对照组中检出的1例HPV23相似,均相当于10-2～10-3拷贝.结论 某些皮肤Bowen病的发病与黏膜型HPV密切相关.     

Detection of human papillomavirus type 58 in polydactylous Bowen's disease on the fingers and toes of a woman - concurrent occurrence of invasive vulval and cervical carcinomas

The group related to human papillomavirus (HPV) type 16 (HPV-16, -31, -33, -35, -52, -58 and -67) is dominantly identified in cervical intraepithelial neoplasia and cervical carcinomas. HPV-16 has also been frequently detected in Bowen's disease on the hands and feet. We describe herein a case of polydactylous Bowen's disease on the fingers and toes of a woman who had had radical vulvectomy and hysterectomy for concomitant invasive vulval and cervical carcinomas. All the lesions, except for the lesions on the periungual side of her left index, middle and ring fingers, harbored HPV-58 DNA with more than 100 entire viral genome copies per cell detected by Southern blot hybridization. The histological localization of the viral DNA was confirmed in all the lesions by in situ hybridization. We could also retrospectively demonstrate HPV-58 DNA in her invasive vulval and cervical carcinoma tissues.     

Bowen''s carcinoma of the scrotal skin associated with human papillomavirus type 82

We have previously cloned human papillomavirus type 82 (HPV-82) from a vaginal intraepithelial neoplasia, but it is not known whether HPV-82 can induce a cutaneous lesion. A large erosive nodule developed on the scrotum of a 50-year-old Japanese patient. Histopathologically, the lesion was composed of two distinct parts; one part showing changes characteristic of Bowen's disease in the epidermis, and the other showing elongated rete ridges and proliferation of atypical basaloid cells in the dermis. These parts were partially connected, giving the diagnosis of Bowen's carcinoma. Immunohistochemically, HPV capsid antigen was detected only in the nuclei of a few cells on the upper part of the epidermis. HPV-82 was identified in the lesion by blot hybridization and viral DNA was demonstrated in the lesion by in situ hybridization. HPV-82 has tropism for both the skin and the genital regions.     

Human papillomaviruses of the mucosal type are present in some cases of extragenital Bowen's disease

BACKGROUND: Bowen's disease in the genital area is generally considered to be caused by mucosal high-risk human papillomaviruses (HPVs). However, the detection rate and spectrum of HPVs in extragenital Bowen's disease are various and it is not clear to what extent HPV is involved in its pathogenesis. OBJECTIVES: To assess the degree of association of HPV in extragenital cases by examining detection rates, types, quantities and localization of HPV. METHODS: A polymerase chain reaction (PCR) approach that we had previously established, which can give sensitive detection of a broad range of HPVs from cutaneous [including epidermodysplasia verruciformis-related HPVs (EV-HPVs)] to mucosal HPVs, was applied to samples from 41 patients with extragenital Bowen's disease and normal skin samples from 48 individuals. Semiquantitative L1-PCR and tyramide-based in situ hybridization (ISH) were also employed for positive cases. RESULTS: HPVs belonging to the mucosal high-risk group were detected in three patients with Bowen's disease (7%; two HPV 16 and one HPV 33), with 10(1)-10(3) copy equivalents per diploid amount of cellular DNA. They were distributed among most nuclei of tumour cells but in none of the cells of adjacent normal skin. HPVs belonging to the cutaneous group were detected in two patients (5%; HPV 27 and HPV 76) at 10(-2)-10(-3) copy equivalents, the same level as in a normal skin specimen positive for type 23 EV-HPV. No positive signals were observed by ISH. CONCLUSIONS: HPVs belonging to the mucosal high-risk group may participate in the development of extragenital Bowen's disease.     

Vulval intraepithelial neoplasia and periungual Bowen's disease concordant for mucosal (HPV-34) and epidermodysplasia verruciformis (HPV-21) human papillomavirus types

Human papillomavirus (HPV) infection is associated with genital malignancy and specific cutaneous malignancies. We report a case of an HPV-associated concurrent vulval intraepithelial neoplasia and periungual Bowen's disease in a young immunocompetent Afro-Caribbean woman with no known risk factors for either disease. HPV genotyping studies detected multiple alpha and beta papillomaviruses with concordance for HPV-34 [a high-risk (HR) mucosal type], and HPV-21 [an epidermodyslasia verruciformis (EV) type] in both vulval and finger tissue. Although the HR-mucosal viruses detected are likely to have a pathogenic role in vulval intraepithelial neoplasia, this is the first report of concordance for EV HPV types in both genital and nongenital skin premalignancies. This case, in the context of accumulating epidemiological and experimental data in cutaneous SCC, raises the question of whether EV HPV may contribute to vulval malignancy, and further study is merited.     

Detection and sequences of human papillomavirus DNA in nongenital seborrhoeic keratosis of immunopotent individuals

BACKGROUND: The etiology of seborrhoeic keratosis (SK) is unknown. Its clinical and histopathological similarities to verrucae vulgaris and condyloma acuminatum prompted us to examine whether human papillomavirus (HPV) is present in SK lesions. In the present study, HPVs were frequently detected from genital lesions or hair follicle in immunocompromised host. OBJECTIVE: We analyzed 104 nongenital SK specimens diagnosed by clinical and histopathological examinations for HPV DNA in immunopotent individuals. METHOD: We analyzed SK specimens for HPV DNA using in situ hybridization (ISH), polymerase chain reaction (PCR), Southern blot hybridization, and sequencing of viral DNA of PCR-amplified fragments. And we also examined virion, which is the capsid protein of HPV in ISH-positive specimens by immunochemical examination. We identified eight mucosal and two cutaneous type HPVs. RESULT: ISH revealed that 30 of 104 (28.8%) SK samples contained HPV DNA. All ISH-positive specimens were demonstrated virion in the nuclei of the epidermal keratinocytes. PCR analysis showed that 87 (83.7%) samples contained HPV-18, 81 (77.9%) HPV-6, and 73 (70.2%) contained both HPV-18 and -6. The incidence of HPV-1 (7.7%) and HPV-2 (14.4%) was relatively low. All 20 normal controls were negative for HPV DNA by ISH but seven were positive by PCR sequencing. CONCLUSION: Our results suggest that HPV, possibly coinfection with HPV-6 and -18 and unknown type(s) of HPV, plays an important role in the pathogenesis of SK.     

Oncogenic human papillomaviruses in extra-genital Bowen disease revealed by in situ hybridization]

BACKGROUND: The association between mucosal oncogenic human papillomaviruses (HPV) and bowenoid papulosis or genital Bowen's disease is well documented. In contrast this association with extra-genital Bowen's disease is poorly studied. The aim of this study was to detect oncogenic (16/18, 31/33/51) and non oncogenic (8/11) mucosal HPV using a in situ hybridization method in 28 skin biopsy specimens of extra-genital Bowen's disease.PATIENTS AND METHODS: Twenty-eight cases of extra-genital Bowen's disease seen in the period 1990-96 in the Dermatology department were included: 19 women and 9 men (mean age: 72 years). Bowen's disease locations were: hands and feet (8 cases), limbs (11 cases), face (8 cases), trunk (1 case). Blinded histopathologic examination confirmed the diagnosis of Bowen's disease and signs of HPV infection (koilocytosis). In situ hybridization was performed using three biotinylated probes detecting HPV types 6/11, 16/18, 31/33/51.RESULTS: Oncogenic HPV genoma was detected in 8 skin samples (28.6 p. 100). In all these cases, 16/18 probe was positive and in two cases, both 16/18 and 31/33/51 probes were positive; 4/8 Bowen's diseases of the extremities were positive for HPV. Koilocytes were found in 6/8 of skin samples with positive HPV detection.DISCUSSION: Mucosal oncogenic HPV are detected by in situ hybridization in 28.6 p. 100 of extra-genital Bowen's disease. In situ hybridization is an easier technique than Southern-Blot hybridization which is the gold standard. Five studies reported similar results and three studies reported different results that we discuss. A precise understanding of oncogenic HPV implication in the development of extra-genital Bowen's disease could lead to the development of new therapeutic strategies (topical cidofovir or imiquimod).     

Premalignant lesions and cancers of the skin in the general population: evaluation of the role of human papillomaviruses

To evaluate the role of human papillomaviruses (HPV) in the development of premalignant lesions and cancers of the skin in the general population, 314 biopsies obtained from 227 patients with benign neoplasms, premalignant lesions, and cancers of the skin and from 25 patients with squamous cell carcinoma of the lip were analyzed by Southern blot hybridization. DNA probes specific for various cutaneous and genital HPV types were used in hybridizations conducted under nonstringent or stringent conditions. HPV DNA sequences were only detected in eight specimens obtained from six patients: HPV 34 in one case of periungual Bowen's disease, HPV 36 and an as yet uncharacterized HPV in two cases of actinic keratosis, HPV 20 in one case of basal cell carcinoma, an as yet unrecognized HPV in one case of squamous cell carcinoma, and HPV 16 in one case of squamous cell carcinoma of the lip. None of the specimens of cutaneous horn and keratoacanthoma contained detectable HPV DNA. In contrast, HPV DNA sequences, mostly HPV 16, were detected in 13 of 23 cases of anogenital Bowen's disease and invasive Bowen's carcinoma. HPV DNA sequences were not detected in 90 cutaneous samples further analyzed by the polymerase chain-reaction technique, using amplification primers that contain conserved sequences among the genomes of HPV. These results strongly suggest that the known HPV types play only a minor role, if any, in skin carcinogenesis in the general population.     

Human papillomavirus-16 DNA in a cutaneous invasive cancer

A middle-aged man with a 20-year history of a warty lesion on his finger was diagnosed histologically as having an invasive carcinoma at that location. Molecular analysis of the lesion showed that human papillomavirus (HPV) DNA was present in this tumor. Under stringent hybridization conditions, only HPV-16 DNA, which is commonly associated with genital neoplasia or cutaneous bowenoid lesions, was detected. Two-dimensional gel electrophoresis revealed that various sections of the tumor contained mostly episomal viral DNA; but in one case, low levels of integrated HPV-16 DNA were detected as well.     

Papillomavirus infection of the anogenital region: correlation between histology, clinical picture, and virus type. Proposal of a new nomenclature

The clinical and histologic picture of 84 anogenital condylomatous and condyloma-like lesions of both sexes were analyzed in an effort to establish a correlation to the different papillomavirus (PV) types. The presence of human papillomavirus (HPV)-specific DNA sequences was confirmed through molecular hybridization and the presence of PV structure antigens was verified in thin sections by means of a group-specific anti-PV-antiserum using the peroxidase-antiperoxidase (PAP) technique. Three distinct clinical forms harboring distinct HPV types were distinguished: (1) Condylomata acuminata in which HPV-6 DNA was present in 37 of 59 samples and HPV-11 DNA in only 13 of 59 samples. HPV-16 DNA was not detected at all and 9 condylomatous lesions remained unclassified. (2) Flat condyloma-like lesions, where HPV-6 and HPV-11 were associated with lesions of low epidermal atypia in 8 and in 2 of 18 cases, respectively, and where HPV-16 was associated exclusively with 6 of 18 such lesions with severe atypia, called bowenoid papulosis. (3) Pigmented papules where HPV-16 was detected twice in lesions of bowenoid papulosis and HPV-11 in 2 of the benign pigmented lesions. The fourth clinical manifestation of genital papillomavirus infections--the so-called condylomata plana--was not available for virologic analysis. Histologically 5 different koilocytotic features were determined which could not be correlated either with one of the clinical pictures or with a specific PV type. HPV-16, however, was found frequently in non-koilocytotic lesions exhibiting the features of severe epithelial atypia known in bowenoid papulosis. The existence of PV structure antigens in these lesions could not be verified using the indirect immunoperoxidase--PAP-technique--in contrast to the koilocytotic lesions where clear evidence of the presence of HPV was proved in 36 of 56 (64.3%) of the cases.     

Characterization and analysis of human papillomaviruses of skin warts

We analysed human papillomavirus (HPV) infections in 61 tissue specimens of skin warts of Taiwanese patients by DNA hybridization. The prevalence of HPV infection was 69% by Southern blot hybridization. The typing of HPVs was performed by dot blot hybridization under highly stringent conditions with each probe separately. The prevalence of HPV-1, 2/3, 4, 5, 8, 11, 16 and 18 in skin warts was 13, 7, 16, 2, 0, 5, 2 and 8%, respectively. Chi-squared analysis revealed that there was a correlation between HPV type and copy number. Most HPV-4-induced warts were verruca vulgaris. HPV-1 DNA was detected in verruca plantaris and verruca vulgaris. No specific histopathological features were found to be indicative of the presence or absence of HPV, or of the various types of HPV infection.     

Epidermodysplasia verruciformis associated with Bowen's carcinoma, B lymphocytopenia and decreased immune functions

A case of epidermodysplasia verruciformis associated with Bowen's carcinoma, persistent B lymphocytopenia and decreased immune functions is reported. Human papilloma virus 5 (HPV-5) DNA was shown to be associated with the DNA from the tumor tissue by Southern blot hybridization using P-labeled HPV DNA sequences cloned on plasmid vectors. The associated Bowen's carcinoma in our case may be caused by multiple-factor relationships which include (a) an oncogenic potential of infected virus; (b) an inherited abnormality in immune function, and (c) the decreased immune function resulting from the viral infection itself. A marked B lymphocytopenia appears to be associated with persistent viral infection.     

Apoptosis in Bowen's disease. An ultrastructural study

We performed ultrastructural studies of apoptosis (previously referred to as "malignant dyskeratosis") in a case of genital Bowen's carcinoma in which human papillomavirus (HPV) type 33 genome was identified and in two cases of cutaneous Bowen's disease with no detectable viral DNA; herein we present the sequential stages in the development of apoptotic bodies. The apoptotic process in the HPV-containing genital Bowen's disease was similar to that in the cutaneous lesions with no detectable HPV. The presence of a large number of apoptotic bodies in Bowen's disease may be responsible for the slow progression and noninvasive growth of this carcinoma in situ.     

Human papillomavirus-32-positive extragenital Bowenoid papulosis (BP) in a HIV patient with typical genital BP localization

OBJECTIVES: Bowenoid papulosis (BP) is characterized by multiple maculopapular lesions of the genitalia; extragenital localization is rarely found. Human papillomavirus (HPV) type 16 has been found in most cases of BP, but the other HPV genotypes associated with BP had been poorly characterized. GOAL: We describe an extragenital BP with concomitant genital involvement in an HIV-positive woman. STUDY: Several HPV gene-specific amplifications and sequencing were performed on DNA extracted from biopsy samples. RESULTS: The presence of HPV-16 DNA in anogenital and HPV-32 DNA in lip lesions was demonstrated in the absence of any other HPV type and with no coinfection. Semiquantitative polymerase chain reaction analysis revealed that HPV-16 and -32 DNA sequences are in the episomal state. CONCLUSIONS: This is the first report of BP associated with HPV-32 (the causative agent of a benign hyperplasia) and a rare case of BP associated with 2 HPV types in the same patient. The immune depression could have modified the normal benign progression of the extragenital lesions.     

Genital tract papillomavirus infection in children

Genital tract papillomas in five children were examined for the presence of human papillomavirus (HPV) DNA by molecular hybridization. Papillomavirus DNA was detected in each sample and was identified as HPV-6 (three cases), HPV-6 or HPV-11 (one case), or HPV-16 (one case). These viruses are the same as are responsible for genital papillomas (condylomata) of adults. The transmission of adult genital tract viruses to children occurs primarily by a venereal route but may occur by a nonvenereal route.     

Detection of human papillomavirus in nongenital Bowen's disease by in situ DNA hybridization

Genital Bowen's disease has been strongly linked in recent studies to human papillomavirus (HPV). Nongenital Bowen's disease has been less well investigated, although isolated reports, all of which involved detection of HPV after extraction of DNA from fresh-frozen tissue, have been made. We investigated 25 cases of nongenital Bowen's disease in 5 black and 20 white patients for the presence of HPV types 1, 6, 11, 16, and 18 using paraffin-embedded tissues. Human papillomavirus was present in six specimens from 3 of the 5 black patients (one previously reported to be positive on Southern blot) and 3 of the 20 white patients; HPV 16 was detected in all 6 cases on low-stringency testing, but only 4 remained positive on high-stringency testing. This suggests an HPV closely related to but not entirely homologous with HPV 16 in the 2 remaining cases. Five of the 6 positive specimens were lesions from the hands and feet and 1 was from the volar aspect of the arm. Clinical factors associated with the presence of HPV included black race, location on the palmar surface and the feet, young age, and verrucous or hyperkeratotic clinical appearance. Of the 6 positive cases, all 5 of the patients available for examination also had evidence of HPV-associated genital lesions. No specific histopathologic features were found to be indicative of the presence or absence of HPV.     

Isolated extragenital HPV-thirties-group-positive bowenoid papulosis in an AIDS patient

We report a case of extragenital bowenoid papulosis in a man with AIDS. The lesions occurred on the anterolateral aspects of the neck, and were not associated with clinical genital or periungual involvement. In situ hybridization demonstrated abundant HPV DNA within the thirties group (31/33/35), in the absence of HPV-16 or -18.     

尖锐湿疣HPV导流杂交快速基因分型及临床意义

目的：探讨外生殖器及子宫颈尖锐湿疣患者皮损脱落细胞或组织中人乳头瘤病毒(Human PapillomaVirus，HPV)9种常见的基因亚型感染分布状况及临床意义。方法：采集患者生殖器或子宫颈尖锐湿疣组织或脱落细胞样本，分离DNA，PCR扩增后与9种常见HPV亚型探针进行导流快速反斑点印迹杂交检测HPVDNA。结果：经PCR扩增并通过通用探针杂交分析后，108例患者标本中，101例标本呈HPVDNA阳性，7例HPV阴性。HPVPCR阳性标本中，98例成功分型。5种高危型HPV16，18，3l，33，58的检出率分别为31．5％(34／108)、16．7％(18／108)、6．5％(7／108)，4．6％(5／108)，13．0％(14／108)。三种低危型HPV6，11，53型的检出率分别为41．7％(45／108)，56．5％(61／108)和6．5％(7／108)。危险度待定HPV亚型CP8304检出率为4．6％(5／108)。结论：结论HPV11、6、16型感染是尖锐湿疣的主要型别。     

原位杂交检测鲍温样丘疹病及Bowen病中HPV16 DNA

采用生物素标记的核酸探针原位杂交技术,检测了30例鲍温样丘疹病、15例Bowen病中16型人乳头瘤病毒(HPV16)感染的组织定位及染色模式。结果显示,30例鲍温样丘疹病中,HPV16阳性13例;15例Bowen病中,HPV16阳性6例。鲍温样丘疹病、Bowen病中HPV16感染累及棘细胞全层;主要为核内团块状着色。Bowen病中,HPV16感染尚累及基底层细胞且存在HPV的点状染色模式。提示鲍温样丘疹病与Bowen病比较,HPV16感染的组织定位及染色模式均有所不同。