Prospective study on the development of antibodies against human papillomavirus type 6 among patients with condyloma acuminata or new asymptomatic infection

The development over time of antibodies to human papillomavirus type 6 (HPV 6) was studied prospectively in a group of 26 condyloma patients receiving multiple treatments, as well as in 17 healthy subjects with a new HPV 6 infection. The condyloma patients provided serum samples during 2–12 visits over a period of 1–20 months, and the levels of IgA, IgG and IgM antibodies to HPV 6, 11 and 16 capsids, and to HPV 6-derived peptide antigens were determined. Among the capsid antibodies, the IgG and IgA responses against HPV 6 were the most prominent, whereas IgM was detected only in a few patients. During follow-up, seroconversion to HPV capsids was found in six patients and antibody peaks disappeared during follow-up in nine patients. There was no association between serology and cure of the wart. One hundred twenty subjects who reported multiple sexual partners, but found negative for HPV DNA at multiple anogenital and oral sites, provided serum samples and swabs for HPV DNA analysis at 2–7 visits during 4–74 months. Seventeen subjects acquired HPV 6 DNA and were included in the study. Serum IgA, IgG and IgM antibodies to HPV 6 capsids were induced concomitantly with the acquisition of viral DNA (P = 0.02, 0.002 and 0.006, respectively), and declined again when the viral DNA was no longer detectable. In conclusion, antibodies to HPV 6 did not predict the clinical course of condylomatous disease, but appear to be useful mainly as epidemiological markers of viral exposure. © 1995 Wiley-Liss, Inc.     

Overexpression of the proto-oncogene C-jun in association with low-risk type specific human papillomavirus infection in condyloma acuminata

Infection with different types of human papillomavirus (HPV) is associated with neoplasia at different anatomic sites. The “low-risk” HPVs (LR-HPV) are responsible for benign genital lesions such as condyloma acuminata. In order to clarify the tumorigenic mechanism of LR-HPV, the HPV infection status was investigated and the expression of the c-jun proto-oncogene in different HPV-related skin and genital lesions analyzed. Of the 17 condyloma specimens analyzed by Western blotting, 13 cases (76.5%) exhibited overexpression of the c-jun gene. All 13 cases harbored high copy numbers of the LR-HPV genome with an average of 926 copies per cell, whereas the other four cases had an average of 12 copies of LR-HPV per cell (P < 0.001). Further typing of HPV by Southern blotting revealed that HPV-6 and HPV-11 infections predominated in c-jun positive cases. The c-jun protein was detected much less frequently in cervical cancers (three of 29, or 10.3%) and skin warts (one of 10), and was not detected in five genital polyps or in five normal cervical tissues. These findings suggest a type 6/11-specific induction of c-jun gene expression in HPV-related neoplastic lesions. © 1996 Wiley-Liss, Inc.     

Structural analysis of human papillomavirus type 6c isolates from condyloma acuminatum and juvenile-onset and adult-onset laryngeal papillomata

The human papillomavirus type 6c (HPV-6c) genome was molecularly cloned from biopsy specimens of a juvenile-onset and an adult-onset respiratory-tract papillomata and a condyloma acuminatum of the cervix. To determine if the genital-tract isolate and respiratory-tract isolates contain divergent sequences that may account for a difference in tissue trophism or for a difference in the age of onset of the disease, fine-structure mapping, heteroduplex analysis by electron microscopy, and nucleotide sequencing were used to examine the sequence relationship among these HPV-6c isolates. No differences were found in the digestion among these HPV-6c isolates. No differences were found in the digestion patterns with 23 restriction enzymes. Heteroduplex analysis among the three genomes demonstrated that they were colinear without apparent deletions or rearrangements and had greater than 90% sequence identity. In heteroduplex analyses with a different subtype (HPV-6e) that was molecularly cloned from a genital wart, the genomes were colinear with greater than 90% sequence identity over 90% of their length. The most divergent region had 75–80% sequence identity and was localized to the part of the genome containing the E5a and E5b open reading frames (ORFs). Comparison of the sequence of 1430 nucleotides in this region for two of the HPV-6c isolates did not identify any differences between them. Comparison with the published sequences of HPV-6b identified deletions/insertions and base changes with approximately 75% sequence identity, and comparison with HPV-11 identified only six base changes. Conservation of sequences in the E4-E5 region and similarity in the restriction enzyme maps demonstrated that HPV-6c and HPV-11 are independent isolates of the same HPV-6 subtype.     

尖锐湿疣防治的免疫学研究进展

关于尖锐湿疣预防和治疗的免疫学研究,近年来多集中在患者的免疫学变化及其与发病和复发的关系、局部治疗与免疫调节剂的使用以及在预防方面着重研制有效疫苗等方面.     

Variation of human papillomavirus type 6 (HPV-6) and HPV-11 genomes sampled throughout the world.

We examined the genomic diversity of human papillomavirus type 6 (HPV-6) and HPV-11 isolates from different parts of the world by comparing the nucleotide sequences of part of the long control region of three reference clones and 62 HPV-6 and 40 HPV-11 isolates from Africa, Europe, Asia, and North and South America. The genomic sequence of the HPV-6b reference type had to be amended by inclusion of a 94-bp segment, which is also present with minor differences in HPV-6a. Aside from two small inserts typical of all variants related to HPV-6a and three inserts found in HPV-11 variants, no major alterations to the size of the long control regions of these viruses were observed. This corrects the previous impression that these two HPV types are highly polymorphic. Altogether, 19 HPV-6 and 10 HPV-11 variant genomes could be distinguished, and most of the differences were due to point substitutions. The variants of either type were continuously connected in phylogenetic trees rather than clustered separately into subtype groups. Thirteen mutations, namely, the two HPV-6a inserts and 11 substitutions in HPV-6 or HPV-11 variants, reduced the dissimilarity between the types, but they bridged only a small fraction of the genetic distance between the two types. Genomes more obviously intermediate between HPV-6 and HPV-11 were not found and probably do not exist any more.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of genital tract papillomaviruses HPV-6 and HPV-16 in warts of the oral cavity

Warty lesions of the oral cavity were examined for etiologic association with genital tract papillomaviruses HPV-6, HPV-11, and HPV-16. DNAs extracted from ten oral biopsies were screened for HPV genomic sequences by Southern transfer hybridization with 32P-labeled viral DNA probes. Nonstringent hybridization with an HPV-6 probe revealed papillomavirus DNA sequences in four of seven tissues with histologic evidence of papillomatosis, in none of two tissues without histologic evidence of papillomatosis, and in one tissue that was not examined by histology. Stringent hybridization tests with HPV-6 and HPV-16 probes identified the genome in one tissue as being HPV-16, in a second tissue as being HPV-6 subtype a, and in a third tissue as HPV-6 (subtype unidentified); papillomavirus DNA sequences in two tissues are as yet not identified. An additional case of HPV-6 or HPV-11 related oral cavity lesion was diagnosed by in situ hybridization of paraffin sections with a 35S-labeled, mixed HPV-6 + HPV-11 probe. The hybridization in the positive section was extensive and confined to epithelial nuclei. The oral lesions associated with genital tract papillomaviruses were asymptomatic, multiple or single, and were located in different parts of the oral cavity, for example, on the gingivae, on the tongue, on the lip, on the tonsillar pillar, and on the floor of the mouth.     

咪喹莫特乳膏对尖锐湿疣免疫调节作用及复发率的影响

目的 探讨采用咪喹莫特乳膏对尖锐湿疣免疫调节作用以及复发率的情况.方法 将尖锐湿疣患者80例随机分为观察组和对照组,各40例.对照组给予激光治疗;观察组在激光治疗后给予咪喹莫特乳膏治疗.结果 观察组治疗总有效率为92.50％,对照组患者治疗总有效率为80.0％,观察组治疗后复发率为12.50％,而对照组复发率为40.00％,差异具有统计学意义(P＜0.05).治疗后,观察组患者IL-4和IL-10水平低于对照组患者IL-4和IL-10水平,且观察组患者IL-12和IFN-γ水平高于对照组患者IL-12和IFN-γ水平,差异具有统计学意义(P＜0.01).治疗后观察组患者CD +4、CD +4/CD +8、NK水平均高于对照组患者CD +4、CD+ 4/CD+8、NK水平,差异具有统计学意义(P＜0.01).观察组患者CD+8水平低于对照组患者CD +8水平,差异具有统计学意义(P＜0.01).治疗6个月后,观察组尖锐湿疣复发患者的CD+3、CD+4、CD+ 4/CD+8以及NK水平低于未复发患者CD+3、CD+4、CD+ 4/CD+8以及NK水平;且复发患者的CD +8水平高于未复发患者CD +8水平,差异具有统计学意义(P＜0.05).结论 咪喹莫特乳膏治疗尖锐湿疣疗效显著,可有效降低复发率,提高患者细胞免疫调节作用是防止尖锐湿疣复发的重要机制.     

Association of E6AP (UBE3A) with human papillomavirus type 11 E6 protein

The cellular E3 ubiquitin ligase E6AP (UBE3A) interacts with the cancer-associated HPV E6 oncoproteins, where together with the viral E6 oncoprotein it binds and targets the degradation of the p53 tumor suppressor. We find that the HPV-11E6 protein also associates with E6AP in vivo, and thereby can target the degradation of an E6-associated protein. Mutation of an E6-binding LXXLL peptide motif on E6AP eliminated the association, revealing a common mode of interaction between high- and low-risk E6 proteins and E6AP. E6AP was required for the in vivo degradation of DLG1 by both HVP-18 E6 and a chimeric HPV-11E6. The common functional interaction of both cancer-associated and non-cancer-associated E6 proteins with E6AP establishes a common mechanism for E6 proteins trophic to mucosal squamous epithelium.     

Genomic characterization of a novel human papillomavirus (HPV-117) with a high viral load in a persisting wart

Warts from immunosuppressed organ transplant recipients (OTR) persist over years and may progress into non-melanoma skin cancer. Human papillomaviruses (HPV) are considered the causal agents for the development of such warts. We isolated the novel type HPV-117 from a persisting wart by rolling circle amplification. One hundred eighteen warts from immunocompetent patients (IC) and 49 warts from OTR were analyzed by HPV-117 E6 type-specific PCR. As inferred from a phylogenetic analysis, the new type HPV-117 belonged to alpha-PV species 2, including the most similar types HPV-10 and HPV-94. The general prevalence of HPV-117 in warts was 2% in IC (2/118), and 12% in OTR (6/49). The high viral load in dysplastic cells of a Verruca vulgaris was shown by in situ hybridization. Our results suggest an active role of the novel type in the development of cutaneous warts of OTR.     

High-grade dysplasia in genital warts from two patients infected with the human immunodeficiency virus

Cancer-associated human papillomavirus (HPV) types are detected in genital warts removed from immunosuppressed individuals more commonly than from those occurring in otherwise healthy individuals. The prognosis of genital warts containing cancer-associated HPV types is not known. Because it is assumed that genital warts are benign lesions, they are usually treated by destructive therapies without prior knowledge of histopathology. The aim of the present study was to determine whether genital warts from individuals with or without human immunodeficiency virus (HIV) contain high-risk HPV types or areas of dysplasia. The study design was a nonrandomized analysis of genital warts removed by excision biopsy from 15 HIV-infected patients and 15 HIV-negative patients. The tissue was analyzed for HPV DNA by hybrid capture, and microscopic sections of each biopsy were examined for areas of dysplasia. Genital warts from HIV-infected patients contained cancer-associated (“high risk”) HPV types in 9 of 15 cases, including 1 that contained only a high-risk type. High-grade dysplastic abnormalities were present in 2 of the 15 lesions from this group, both of which contained high-risk HPV types. Four genital warts removed from HIV-negative patients contained high-risk HPV types, but none contained dysplastic abnormalities. It is concluded that genital warts from HIV-infected patients often contain high-risk HPV types. Such lesions may exhibit dysplastic changes. The frequency of dysplastic changes in genital warts from HIV-infected patients is not known. Biopsy of genital warts may be indicated prior to additional therapy in HIV-infected patients, and surgical removal should be considered as a preferred treatment option in these patients. J. Med. Virol. 54:69–73, 1998. © 1998 Wiley-Liss,Inc.     

Antibody response to human papillomavirus (HPV) type 11 in children with juvenile-onset recurrent respiratory papillomatosis (RRP).

We previously established, using an ELISA, the presence of specific antibodies directed at human papillomavirus (HPV) type 11 virions in the sera of patients with condylomata acuminata, mostly a disease of young adults that, like recurrent respiratory papillomatosis (RRP), is caused by two closely related HPVs, types 6 and 11. The present study was done to investigate if children with RRP can make viral-specific antibodies to an infection that is acquired at birth. Using the same ELISA, we studied the sera of 32 children with biopsy-documented juvenile-onset RRP and compared them to the sera of 31 control children. The median (and interquartile range) of the OD values in the controls and the cases was 0.078 (0.003, 0.101) and 0.230 (0.063, 0.725), respectively, a statistically significant difference (P = 0.001). Among the cases, there was no difference in seroreactivity between children with HPV-11-induced RRP and those with HPV-6-induced RRP (P = 0.31). Since HPV-11 viral particles do bind to the ELISA plate and remain intact and accessible to antibodies, we conclude that children with RRP, like adults with condylomata acuminata, develop antibodies directed at HPV-11 virions.     

Analyses of human papillomavirus genotypes and viral loads in anogenital warts

Condylomata acuminata (genital warts) are the most common sexually transmitted viral diseases. These lesions are caused by infection with mucosal human papillomaviruses (HPVs). However, there is limited information on HPV strain distribution involved in the molecular pathogenesis of these lesions. To address this, the strain prevalence and the frequency of multiple HPV infections were determined in wart tissue obtained from 31 patients attending a wart clinic. These lesions were bisected and subjected to parallel DNA and mRNA extractions. HPV-type prevalence and incidence of multiple infections were determined by the Roche Linear Array assay. qPCR compared HPV 6, 11, 16, and 18 viral loads and RT-qPCR measured HPV 6 and 11 E6 genomic expression levels. Seventy-one percent of these samples were infected with multiple HPVs. Only one sample was negative for HPV 6 or 11 DNA. Forty-eight percent of samples were positive for a high risk (oncogenic) HPV. The results show that multiple infections in tissue are frequent and the subsequent analysis of HPV 6 and 11 E6 DNA viral loads suggested that other HPVs could be causing lesions. Further analysis of HPV 6/11 E6 mRNA levels showed that there was no discernable relationship between HPV 6 E6 DNA viral load and relative HPV 6 or 11 E6 mRNA levels thereby questioning the relevance of viral load to lesion causality.     

The seroprevalence of IgG antibodies to human papillomavirus (HPV) types HPV-16, HPV-18, and HPV-11 capsid-antigens in mothers and their children

Human papillomavirus (HPV) types causing anogenital lesions and cancer are accepted as being sexually transmitted. The methods whereby children acquire these anogenital type HPV infections are unclear. The present study determined the prevalence of anti-HPV-16, HPV-11 and HPV-18 IgG antibodies in mothers and their children in an attempt to identify evidence of HPV transmission from mother to child. HPV virus-like particles (VLP) VLP-16, VLP-11 and VLP-18 were used in enzyme-linked immunosorbent assay to identify IgG antibodies in serum from 100 mothers and their 111 children. Antibodies to VLP-16, VLP-11 and VLP-18 were found in serum from 17%, 21% and 16% of mothers, respectively and seroprevalences were 9%, 11.7% and 9.9%, respectively amongst the children. Of the 111 children, 23 (20.7%) showed antibodies to one or more of the three HPV types tested. Seven of these (30.4%) HPV IgG positive children had the same antibodies to one or more HPV types as their mothers. The prevalence of HPV-11 was similar in children of seropositive compared with seronegative mothers (14% and 11%, respectively). The prevalence of HPV-16 and HPV-18 was higher in children of seropositive mothers compared with seronegative mothers (for HPV-16, 18% and 7%, respectively, P = 0.1, for HPV-18, 19% and 8%, respectively, P = 0.2). None of these differences were statistically significant indicating a lack of correlation between antibodies in mothers and children and no evidence to support vertical or horizontal mother to child transmission of HPV infection. Indications were of multiple sources of HPV infection in the children.     

Relationship between antigenic type of virus and antibody response in female patients with herpes genitalis

The antigenic types of virus recovered from genital sites in cases of adult female genital herpes and antibody response in these patients were investigated. Twelve strains were isolated from 23 clinical specimens, and half the number of the isolates was classified as type 1 virus and the remaining half as type 2 virus. The results of serological typing corresponded well to biological differentiation by plaque-forming ability on chick embryo cultures. The patients with type 1 virus infection had only the complement requiring neutralizing antibody, thus indicating a fresh infection. On the other hand, the noncomplement requiring neutralizing antibody was found in the patients with type 2 virus infection, indicating a later phase of infection.     

Demonstration of URR-duplication variants of human papillomavirus type 6 in paraffin-embedded tissue sections of one condyloma acuminatum and one buschke-loewenstein tumour

Human papillomavirus type 6 (HPV 6) induces condylomata acuminata and laryngeal papillomas. Occasionally, HPV 6 may also be found in low-grade verrucous carcinomas. In some tumours, genetic analysis revealed the presence of HPV 6 variants with rearrangements, mostly DNA duplications, within the upstream regulatory region (URR). In this study, we analysed 98 formalin-fixed, paraffin-embedded condylomata acuminata obtained from 54 patients for the presence of URR-duplication variants of HPV 6. HPV 6 DNA could be amplified by polymerase chain reaction (PCR) from 40 samples. One condyloma acuminatum contained a HPV 6 genome with rearranged URR. Analysis using restriction enzyme cleavage suggested a DNA duplication within the URR of approximately 200 bp, spanning the Hpa II site at nt 7863 and the Dde I site at nt 7843 but not involving the Rsa I site at nt 7633. In addition, we analysed the distribution of the already characterized URR-duplication variant HPV 6_ACIB within different paraffin-embedded tissue sections of a Buschke-Loewenstein tumour. No correlation could be demonstrated between the presence of the rearranged genome and malignant histological changes. This result and the demonstration of an URR-duplication variant in a typical condyloma acuminatum suggest that duplications within the URR of HPV 6 are not directly related to malignant progression of HPV 6-induced tumours.     

Patterns of antigenaemia and antibody response in patients infected with human immunodeficiency virus (HIV) according to clinical state.

Five hundred and fifty six subjects, known to be homosexuals or intravenous drug abusers and seropositive for HIV antibody, were selected on the basis of their clinical state--symptom free, lymphadenopathy syndrome (LAS), AIDS related complex (ARC), and AIDS. The presence of antigenaemia and the humoral response to viral polypeptides was investigated. The prevalence of patients positive for p31 antibody was significantly increased in those with AIDS and detectable antigenaemia.     

Mapping of human serum-reactive epitopes in virus-like particles of human papillomavirus types 16 and 11

Most human antibodies against HPV16 can be blocked by the monoclonal antibody H16.V5. To investigate whether H16.V5 and human sera recognize similar epitopes, hybrid capsids containing different parts of HPV16 and HPV11 were evaluated for reactivity with human sera. The antibody responses among HPV 16-/HPV11+sera to HPV11 and to hybrid capsids containing the HPV11 C-terminus were strongly correlated. The antibody responses among HPV 16+/HPV11-sera to HPV16 and to a hybrid containing the HPV16 C-terminus were correlated and there was also reactivity with a hybrid containing the H16.V5 epitope in the HPV11 backbone. Several HPV16-/11- children's sera were reactive with hybrid capsids, implying that a native capsid structure is essential for serological specificity. For both HPV16 and HPV11, the major serologic reactivity was directed toward the C-terminal part of the protein and the H16.V5 binding site appeared to be a major serologically reactive epitope of HPV16.