Human Papillomaviruses and genital co-infections in gynaecological outpatients.

BACKGROUND: High grade HPV infections and persistence are the strongest risk factors for cervical cancer. Nevertheless other genital microorganisms may be involved in the progression of HPV associated lesions. METHODS: Cervical samples were collected to search for human Papillomavirus (HPV), bacteria and yeast infections in gynaecologic outpatients. HPV typing was carried out by PCR and sequencing on cervical brush specimens. Chlamydia trachomatis was identified by strand displacement amplification (SDA) and the other microorganisms were detected by conventional methods. RESULTS: In this cross-sectional study on 857 enrolled outpatients, statistical analyses revealed a significant association of HPV with C. trachomatis and Ureaplasma urealyticum (at high density) detection, whereas no correlation was found between HPV infection and bacterial vaginosis, Streptococcus agalactiae, yeasts, Trichomonas vaginalis and U. urealyticum. Mycoplasma hominis was isolated only in a few cases both in HPV positive and negative women and no patient was infected with Neisseria gonorrhoeae. CONCLUSION: Although bacterial vaginosis was not significantly associated with HPV, it was more common among the HPV positive women. A significant association between HPV and C. trachomatis was found and interestingly also with U. urealyticum but only at a high colonization rate. These data suggest that it may be important to screen for the simultaneous presence of different microorganisms which may have synergistic pathological effects.

Detection of human papillomavirus and adenovirus in benign and malignant lesions of the larynx.

OBJECTIVE: To investigate the role of human papillomavirus (HPV) and adenovirus (AdV) infections in the oncogenesis of squamous cell carcinomas of the larynx and of laryngeal dysplasia. STUDY DESIGN: Cross-sectional study with planned data collection. SETTING: Department of Otorhinolaryngology G. Ferreri and Department of Experimental Medicine and Pathology-Section of Virology, Sapienza University of Rome. SUBJECTS AND METHODS: Biopsy samples were taken from 68 patients with benign and malignant lesions of the larynx. All tissue samples were analyzed by means of polymerase chain reaction with two groups of primers for HPV and with a pair of primers for AdV. RESULTS: All cases of carcinomas and dysplasia as well as all control cases were negative for both viruses. Four of the five cases of laryngeal papillomas were positive for only HPV, confirming the role of these viral types in the origin of papillomas. CONCLUSION: The absence of viral genomes in laryngeal carcinomas as in the other cases studied suggests the existence of other factors that play a more important role than viral infection in the carcinogenesis of these lesions.

Intra-anal condyloma: surgical or topical treatment?

Human Papillomavirus infections are the strongest risk factors for genital cancer and are the causative agents of anogenital warts. Although the viral types associated with condylomata usually do not cause carcinoma, in women with a history of these lesions there is an increased risk of intraepithelial neoplasia and cancer. Generally the lesions are not life-threatening, but they provoke significant morbidity, are difficult to treat, and are a source of psychosocial stress. Thus, condylomata represent not only a health problem for the patient but also an economic burden for the society. Considering the individual episodes of care, men experience a longer duration of the lesions and incur greater costs than women. We report a case of a male patient with external and intra-anal condyloma resistant to laser therapy. Initially, surgical intervention appeared required because of florid and intra-anal growth. HPV DNA testing and sequencing revealed the presence of HPV 6. After initial discomfort, the lesions were successfully cleared with topical imiquimod 5 percent cream therapy.

Direct sequencing of HPV DNA detected in gynaecologic outpatients in Rome, Italy.

In order to assess the frequency of different human papillomavirus (HPV) types in Rome and the association between HPV and behavioural characteristics, we tested cervical scrapes of a population of sexually active women referring to university clinics for routine gynaecologic care. The presence of HPV DNA was revealed by polymerase chain reaction on two genome regions (L1 and E6/E7) followed by sequencing. Thirty different HPV types were identified; HPV 16 was the most prevalent (14.18%), followed by HPV 53 (9.21%), HPV 58 (7.80%), HPV 6 and 66 (both 5.67%) whereas all the other genotypes ranged below 5%. In univariate analysis the characteristics significantly associated with HPV DNA detection were the youngest age (P<0.01), the high number of lifetime partners (P<0.001) and the smoking habit (P<0.01). In multiple logistic regression analyses, the characteristics significantly associated with HPV DNA detection remained the younger age and the higher number of lifetime sexual partners. This study may be interesting in order to evaluate the circulation of HPV genotypes in Italy and to add a contribution to anti-cancer vaccine development.

Lactoferrin inhibits early steps of human BK polyomavirus infection.

Lactoferrin, a member of the transferrin family, is a bi-globular iron binding glycoprotein, found in milk, exocrine secretions of mammals, and in secondary granules of polymorphonuclear neutrophiles that plays an important role in the defence against various pathogenic microorganisms. Previous studies in different virus-cell systems showed that lactoferrin is a potent inhibitor of different enveloped and naked virus infection. In this research we studied the effect of lactoferrin on BK polyomavirus, a human naked double-stranded DNA virus responsible for productive, persistent, and latent infections of the urinary tract. Results obtained demonstrate that lactoferrin treatment prevents early steps of BK virus infection in Vero cells, at the level of the adsorption phase, probably through the interaction with capsidic structures, although a lactoferrin-BK virus competition for cell plasma-membrane receptors cannot be ruled out.

Detection of human herpesviruses and polyomaviruses DNA in a group of patients with relapsing-remitting multiple sclerosis.

Multiple sclerosis (MS) is an inflammatory disease of the central nervous system whose pathological features consist of white matter plaques of primary demyelinization and loss of oligodendrocytes. Various risk factors have been associated with MS susceptibility. We have focused this study on different viruses. In particular in the present study we used PCR to search for the genomic DNA of HHV-1, HHV-2, HHV-8, BKV and JCV in urine and peripheral blood mononuclear cells (PBMC) samples from 44 relapsing-remitting MS (RRMS) patients. No viral DNA was found in any urine sample, whereas 29.5% of RRMS PBMC samples were positive. It is suggestive that Human herpesviruses (HHV-1 and HHV-8) were constantly present in all positive samples, indicating that viral agents could contribute to create the demyelination plaques and cause MS.

Role of Human Papillomavirus in the Pathogenesis of Laryngeal Dysplasia.

OBJECTIVE: A synergistic effect between smoking and alcohol intake is the major cause of premalignant and malignant lesions of the larynx, but the risk factors and pathogenesis of the neoplastic transformation in nonsmokers remain poorly defined. The aim of this retrospective study is to establish the relationship between smoking habits and human papillomavirus (HPV) infection in laryngeal dysplasia. STUDY DESIGN: Cross-sectional study. SETTING: Academic university hospital. SUBJECTS AND METHODS: HPV DNA was amplified from 30 paraffin-embedded laryngeal dysplasia tissue specimens by the polymerase chain reaction using 2 groups of different consensus primers (MYO9/MY11 and LCRF1-4, E7R1-4). Fifteen samples were taken from smokers and 15 from nonsmokers. RESULTS: The present investigation failed to demonstrate the HPV genome in all samples of laryngeal precancerous lesions, whereas HPV was detected in 4 laryngeal papilloma samples used as control to confirm the reliability of our method on paraffin-embedded samples. CONCLUSIONS: Although the small number of cases in our series limits the power of our statistical analysis, the absence of viral genomes in the specimens analyzed in this study suggests the lack of a relationship between HPV infection and laryngeal dysplasia in smokers as well as in nonsmokers.

Nasal polyposis: from pathogenesis to treatment, an update.

Nasal polyps (NP) are common benign degeneration of nasal sinus mucosa with a prevalence around 4% in the adult population. The causes are still uncertain but there is a strong association with allergy, infection, asthma and aspirin sensitivity. Histologically, the presence of a large quantity of extracellular fluid, mast cell degranulation and eosinophilia has been demonstrated. Typically the patients show nasal obstruction, anosmia and rhinorrhoea. Nasal endoscopic examination and CT imaging allow evaluation of the disease extension. A combined medical and surgical treatment is recommended for symptoms control in preventing symptomatic NP recurrence. We will review the current knowledge in the pathogenesis and treatment of this complex disease entity.

Diagnosis of anal human papillomavirus infection: polymerase chain reaction or cytology?

OBJECTIVES: To investigate the relationship between human immunodeficiency virus (HIV)-positive and HIV-negative patients engaging in promiscuous behaviors and anal human papillomavirus (HPV) infection diagnosed by polymerase chain reaction (PCR) and cytology. METHODS: Fifty-six HIV-positive patients and 49 HIV-negative patients who engaged in sexually promiscuous behavior were enrolled in the study. We performed cytological exams using the Pap smear and PCR for HPV-DNA detection, with identification of oncogenic strains. The 2001 Bethesda System terminology was used for the cytological exams. We also evaluated the immunologic status of the HIV-infected patients. RESULTS: PCR positivity for HPV-DNA was higher in the group of HIV-positive patients than in the group of HIV-negative patients with a statistically significant difference. In contrast we did not find any statistically significant difference by cytological exam. Oncogenic strains were equally distributed in the two groups. CONCLUSIONS: Our results indicate the importance of the cytological exam for anal HPV screening in the population at high risk of sexually transmitted disease and that HPV-DNA PCR can be used only as adjunct test.

Trichomonas vaginalis infection: risk indicators among women attending for routine gynecologic examination.

OBJECTIVES: Trichomonas vaginalis is one of the most common agents of sexually transmitted disease, but trichomoniasis is still considered as an infection of minor importance. The aim of this study was to investigate the risk indicators for trichomoniasis in women attending for routine gynecologic examination. METHODS: The study was conducted comparing demographic and behavioral characteristics in two groups of 122 women with T. vaginalis infection and 853 uninfected women. RESULTS: Univariate analysis showed a significant increase of T. vaginalis infection among women with older age and multiple sexual partners. In multiple logistic regression analysis, the higher number of lifetime partners and older age remained significantly associated with T. vaginalis infection. CONCLUSIONS: This study suggests that T. vaginalis infection might be mainly related to lifestyle risk factors. Therefore, in our opinion, the diagnosis and treatment of this disease should be encouraged on routine gynecologic examination.

Type-specific human papillomavirus-DNA load in anal infection in HIV-positive men.

OBJECTIVE: To characterize anal human papillomavirus (HPV) infections in terms of genotype prevalence and type-specific DNA load in HIV-positive men. DESIGN: HIV-positive men attending the colo-proctological clinic of a University Hospital in Rome were recruited prospectively from November 2004 to July 2007. HIV-negative outpatients attending the same clinic over the same period were used as a control group. METHODS: Anal brushings were tested for HPV-DNA using polymerase chain reactions and direct sequencing; type-specific HPV-DNA copies were measured in most positive samples. HPV data were correlated with patient HIV status and risk factors. RESULTS: HPV-DNA infection was detected in 81% of HIV-positive men. Almost all homosexual men were HPV-infected. The infection rate in low-risk HPV types was higher than in high-risk types. The spectrum of HPV genotypes was comparable between HIV-positive and HIV-negative men. Numbers of HPV-DNA copies varied greatly between samples but did not differ significantly between HIV-positive and HIV-negative men. In many samples, low-risk (HPV 6, 61, 70, and 74) viral loads were comparable with those of high-risk HPVs. CONCLUSION: Type-specific HPV-DNA copies at baseline appear to be independent of patient immune status and of HPV genotype. HPV genotype risk and viral load should be further evaluated for their potential predictive role in persistence and progression.

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.

Human immunodeficiency virus infection in vitro activates naturally integrated human papillomavirus type 18 and induces synthesis of the L1 capsid protein.

Human papillomavirus (HPV) infections are prevalent in human immunodeficiency virus (HIV)-positive individuals. To highlight the effect of HIV on HPV expression, HPV-18-positive HIV-permissive HeLa-T4 cells were either infected with HIV-1 or treated with Tat or with the cytokines IL-1alpha, IL-1beta, IL-6 and TNF-alpha. The presence of HPV-18 E1 (early) and L1 (late) transcripts was then determined by dot-blot or Northern blot hybridization with E1 and L1 or with genomic HPV-18 DNA probes, respectively. Protein extracts from parallel cultures were challenged by Western blotting with an antiserum raised against an L1-beta-galactosidase hybrid protein. Results indicated that HeLa-T4 cells constitutively express E1 and L1 transcripts. When cells were infected with HIV, the amounts of E1 and L1 RNAs increased with time, followed by the de novo appearance of L1 protein. E1 and L1 transcripts were also increased, in a dose-dependent manner, by treatment of uninfected cultures with Tat or with IL-6, but were not affected by IL-1alpha, IL-1beta and TNF- alpha. Neither Tat nor IL-6 could induce L1 translation. These findings raise the hypothesis that the increase of HPV shedding and of HPV-associated diseases in HIV-infected individuals could be due in part to a direct or cytokine-mediated action of HIV, in addition to the HIV-induced immunodeficiency.

Polyomavirus persistence in lymphocytes: prevalence in lymphocytes from blood donors and healthy personnel of a blood transfusion centre.

BK and JC polyomaviruses (BKV and JCV) are widespread in humans and are thought to persist and reactivate under immune alterations. In addition to the kidney, lymphoid cells have been proposed as a site of latency. However, while this was shown to occur in immunocompromised patients, discordant data were published for healthy humans. To help to solve this issue, an extensive study (231 healthy subjects) was carried out on peripheral blood mononuclear cells (PBMC) from blood donors of two towns and from operators of a blood transfusion centre. To discriminate between past and recent infection, nested PCRs for BKV and JCV non-coding control region (NCCR) and VP1 DNA sequences were carried out. Twenty-two per cent of subjects had BKV NCCR, but only 7% also had BKV VP1, as detected by PCR assays of similar sensitivities; the latter positivity was found to decrease with age. In both towns, the BKV WW archetypal DDP strain, subtype I, was found. Only 0.9% of subjects contained JCV DNA, for both NCCR and VP1. Blood operators presented a statistically significant increased prevalence of BKV NCCR (3. 0-fold) and BKV VP1 (9.4-fold) sequences with respect to blood donors of comparable ages, suggesting the possibility of occupational risk of BKV (re)infection or reactivation. Since the possibility of amplifying BKV VP1 sequences from PBMC of healthy humans is lost with age, this suggests that PBMC are not a site of polyomavirus persistence in healthy individuals and that detection of BKV VP1 DNA in PBMC is probably indicative of recent infection or reactivation.