HIV-1 seroconversion promotes rapid changes in cervical human papillomavirus (HPV) prevalence and HPV-16 antibodies in female sex workers.

The extent to which human immunodeficiency virus (HIV-1) infection impacts on the ability to mount an effective immune response to HPV is unknown, but is relevant in planning HPV vaccine strategies for HIV-1 infected individuals. This longitudinal study investigated changes shortly after HIV-1 seroconversion on cervical HPV types and HPV-16 antibody responses in serum and at the cervix of female sex workers. Typing of HPV DNA from cervical cells was done prior to HIV-1 seroconversion and within 1 year and greater than 2 years after HIV-1 seroconversion. Antibody determinations on serum and cervico-vaginal rinse samples were by HPV-16 virus-like particle-based, enzyme-linked immunosorbent assay. Of 104 women tested, 40 (38.4%) became HIV-1 seropositive (HIV-positive) during the course of the study. Shortly after HIV-1 seroconversion a significant increase in multiple (>1) HPV infection (OR 4.0, 95% CI 1.3-11.9) was observed compared with HIV-1 seronegative (HIV-negative) women and certain changes in HPV type infection. HIV-1 seroconversion resulted in a reduced prevalence of serum HPV-16 IgA and cervico-vaginal IgA and IgG but an increased prevalence of serum HPV-16 IgG. All HIV-positive women had been exposed to HPV-16 as all displayed serum HPV-16 IgG. Serum HPV-16 responses were maintained at a high magnitude in the presence of HPV-16 infection irrespective of HIV infection, but decreased in the absence of HPV-16 infection. In conclusion, HIV-1 seroconversion in sex workers rapidly increased cervical HPV infection and caused a reduced ability to produce cervical HPV-16 antibodies but a continued ability to generate serum IgG antibodies.

The impact of the use of COL-1492, a nonoxynol-9 vaginal gel, on the presence of cervical human papillomavirus in female sex workers.

This study investigated the effectiveness of a nonoxynol-9 (52.5mg, 3.5%), vaginal gel (Advantage S), in the prevention of human papillomavirus (HPV) infection in female sex workers. We showed by HPV DNA determination in cervico-vaginal rinses a significant increase in multiple (>1) HPV infection in HIV-1 seropositive women compared with HIV-1 seronegative women (OR 4.0, 95% CI 1.3-11.9). We also demonstrated a significant increase in multiple HPV infections in HIV-1 seronegative women using nonoxynol-9 compared with HIV-1 seronegative women using placebo (OR 3.5 95% CI 1.0-11.8). We conclude that the use of nonoxynol-9 did not prevent genital HPV infection and could increase the virus' ability to infect or persist.

Detection of HPV 16 and HPV 18 DNA in the blood of patients with cervical cancer.

Persistent infection of the uterine cervix with high-risk human papillomaviruses (HPV) is causally associated with cancer of the cervix. A few studies have reported the presence of HPV DNA in the blood of women with cervical neoplasia. The aim of this study was to determine if HPV DNA could be detected in whole blood of women with a range of cervical pathologies and with HPV 16 or 18 cervical infections and if there is a correlation between cervical lesion grade and the appearance of HPV DNA in the circulatory system. Forty-five women with histologically graded cervical cancer were confirmed to have cervical HPV 16 or 18 infections. Eleven (24.4%) of these women had detectable HPV 16 or 18 in their blood. The HPV types detected in the blood matched those detected at the cervix. No HPV 16 or 18 DNA was detected in the blood of 32 women with pre-cursor cervical lesions or normal cervical pathology but who had cervical HPV 16 or 18 infections. One of 77 women with normal cervical pathology and no cervical HPV infection was positive for HPV 16 DNA in her blood. The results indicate that HPV DNA can be detected in the blood of women with more advanced cervical carcinomas but not in the blood of women with pre-cursor cervical lesions. The results of our study indicate that the role of HPV DNA in the circulatory system appears not be of diagnostic significance and HPV DNA is only detectable in women with more advanced cervical cancers.

High prevalence of HPV 16 in South African women with cancer of the cervix and cervical intraepithelial neoplasia.

Despite the high prevalence of cervical cancer and cervical neoplasias in South Africa, few studies have been performed in this region to establish which human papillomavirus (HPV) types are associated with the development of high-grade cervical intraepithelial neoplasia lesions and cervical cancer. To investigate these prevalence rates, punch biopsies were obtained from 56 women with cervical cancer and 141 women with histologically diagnosed cervical intraepithelial neoplasia 2 or 3 lesions. Nested polymerase chain reaction (PCR) using consensus degenerate PCR primers was performed for the detection of HPV DNA and HPV typing was done by restriction fragment length polymorphism. Forty-seven (94%) of the cervical cancer and 114 (88%) of the cervical intraepithelial neoplasia 2/3 biopsies were positive for HPV DNA. The prevalence rates of the HPV types detected in the cervical cancer biopsies were HPV 16 (82%), HPV 18, (10%), HPV 33 (10%), HPV 31 (2%), HPV 58 (2%), HPV 35 (2%), and HPV 59 (2%). The cervical intraepithelial neoplasia lesions contained HPV 16 (56.6%), HPV 33 (14%), HPV 31 (10.9%), HPV X (7%), HPV 52 (3.9), HPV 58 (3.1%), HPV 35 (2.3%), HPV 18 (1.6%), HPV 11 (0.8%). Five of the nine fragments that were not typed by the RFLP, designated HPV-X, were sequenced to give HPV6 (1/5), HPV 26 (2/5), HPV 68 (1/5), and candHPV 87 (1/5). HPV 58 was detected in one cervical cancer biopsy and four biopsies from cervical intraepithelial neoplasia grade 3 lesions and was shown to be a previously described variant [Williamson and Rybicki (1991) J. Med. Virol. 33:165-171]. In addition, a cervical intraepithelial neoplasia grade 2 lesion was shown to harbour HPV type HAN2294 (cand HPV 87). The results of this study indicate that cervical cancer and cervical intraepithelial neoplasia 2/3 are largely associated with HPV 16 infection in this group of South African women and, therefore, an effective HPV 16 based vaccine should prevent the development of cervical cancer in a large proportion of women from this region of South Africa.

Typing of human papillomavirus in Zimbabwean patients with invasive cancer of the uterine cervix.

BACKGROUND: Cervical cancer affects 1 in 2000 Zimbabwean women. We investigated the type-specific distribution of human papillomavirus (HPV) infection in Zimbabwean women with invasive cervical cancer. METHODS: We conducted a descriptive study on 98 women with invasive cervical cancer. The methods used were a nested polymerase chain reaction (PCR) for amplification of HPV-DNA and restriction fragment length polymorphism (RFLP) to characterize the HPV types. RESULTS: HPV-DNA was identified in 97% of the cases. HPV types 16, 33, 18 and 31 were identified in 61%, 39%, 18% and 4% of the patients, respectively. We typed one case each of HPV types 35 and 58. Multiple HPV infections were present in 24%. All patients (n = 3) with adenocarcinoma of the cervix were infected with the HPV. Patients infected with HPV-16 alone presented at a median age of 46 years while those infected with HPV-33 alone presented at 43 years. However, patients coinfected with both HPV-16 and HPV-33 were between 10 and 13 years older (median age of 56 years) than patients with either HPV-16 or HPV-33 as single infections. These differences were marginally significant (p = 0.08) or significant (p = 0.02), respectively. CONCLUSION: We present the first prevalence data on HPV types in patients with cervical cancer in Zimbabwe and show that, provided appropriate techniques are employed, HPV infection can be identified in a majority of the patients. The distribution of HPV types should be taken into consideration in tailoring locally relevant vaccines against HPV.

Detection of human papillomavirus in urine and cervical swabs from patients with invasive cervical cancer.

Despite the high prevalence of both human papillomavirus (HPV) infections and cervical cancer among Zimbabwean women, the ability to test for HPV infection of the uterine cervix is limited by a lack of an easy sample collection method that does not require gynecological examination. The presence of HPVs in urine and cervical swab samples collected from 43 women who presented with invasive cervical cancer was investigated. HPV detection was done by means of degenerate primers in a nested polymerase chain reaction (PCR). Typing of HPVs was done using restriction fragment length polymorphism (RFLP) analysis. HPV was identified and typed in 98% (42/43) of cervical swabs and 72% (31/43) of paired urine samples. HPV type 16 was the most common (25/42, 59%), followed by types: 33 (13/42, 31%), 18 (6/42, 14%), and 31 (1/42, 2%). Type-specific concordance between cervical and urine samples was high (22/28, 79%). Therefore, the HPV types identified in urine samples in most cases represent the same HPV type infecting the cervical epithelium. The results suggest that urine may be a practical sample for testing of HPV urogenital infection. Further research is required before the detection of HPV in urine can be applied in the routine cervical screening programs.

The use of nested polymerase chain reaction and restriction fragment length polymorphism for the detection and typing of mucosal human papillomaviruses in samples containing low copy numbers of viral DNA.

Mucosal human papillomaviruses (HPVs) that infect the genital area have also been shown to infect the oral cavity. In this study a restriction fragment length polymorphism (RFLP) method was developed on a nested polymerase chain reaction (PCR) product to identify ten high risk HPV types 16, 18, 31, 33, 35, 45, 51, 52, 58 and 59 as well as the low risk HPV 11. HPV DNA was detected in 23/31 (74%) of buccal specimens using a sensitive nested PCR employing degenerate consensus primers (Williamson and Rybicki, 1991). Consensus PCR using the PGMy09/11 primers. was able to detect HPV in only 29% of the specimens that had tested positive using the nested HPV PCR primers. HPV 11 type specific primers detected HPV 11 DNA in only 66% of the specimens showing HPV 11 DNA by means of nested PCR and RFLP. A Genbank search revealed that the PCR primers could detect a wide range of mucosal HPV types including types HPV 70, 72 and 73 which have all been isolated from immunocompromised patients. Of the 23 buccal specimens that were positive for HPV DNA, 13 were single infections, five were dual infections and three were triple infections. The HPV types identified by RFLP were: HPV 11 (18/23), HPV 18 (8/23), HPV 16 (3/23), and HPV 33 (1/23). HPV 13 (2/23) was identified by direct sequencing of the inner amplicon of the PCR product.

Single-cell cytokine analysis allows detection of cervical T-cell responses against human papillomavirus type 16 L1 in women infected with genital HPV.

Specific types of human papillomavirus (HPV) are known to play a causal role in the development of cervical cancer, with human papillomavirus type 16 (HPV-16) identified as the predominant type. Despite this, little is known about cervical immune responses to this pathogen. The aim of this study was to assess the feasibility of cervical cytobrush sampling and single-cell cytokine staining to investigate cervical lymphocyte-specific cytokine responses to HPV-16 antigens. Of eighteen women recruited into the study, five were HPV DNA positive at the cervix (current exposure) and a further five had circulating antibodies to HPV-16 (previous exposure). Cervical lymphocytes, isolated from the five HPV DNA-positive women, two HPV DNA-negative controls, and one woman with circulating HPV-16 antibodies were assessed for HPV-specific responses using intracellular staining for interferon-gamma (IFN-gamma) and interleukin-4 (IL-4). We demonstrate that both CD4(+) and CD8(+) cervical T lymphocytes, harvested from noninfected and infected subjects, produce these cytokines in response to nonspecific stimulation. However, antigen-specific (HPV-16 L1) IFN-gamma production by CD4(+) and CD8(+) cervical T lymphocytes is only detectable in women exposed currently or previously to HPV-16. This is the first time that antigen-specific cytokine responses of mucosal lymphocytes, obtained from a site of HPV infection, have been demonstrated. This finding clearly illustrates the use of intracellular cytokine staining for investigation of low precursor frequency single-cell antigen-specific responses in lymphocytes harvested from mucosal sites with HPV infection.