Cervicovaginal, oral, and serum IgG and IgA responses to human papillomavirus type 16 in women with cervical intraepithelial neoplasia

Oncogenic human papillomaviruses (HPVs) are obligate mucosal pathogens and typically cause localized infections. The mucosal surface of the genital tract also provides the first line of defense against genital HPV infection. Although local antibody production following HPV-infection has been demonstrated, their role in protection from cervical disease is unclear. This study evaluated oral and cervical HPV infection and the associated linkage between HPV-16 oral, cervical and serum antibody responses in 103 women with varying grades of cervical intraepithelial neoplasia (CIN). We found that HPV-16 was the most prevalent cervical HPV infection (30/103, 29.1%) but was only detected in 1.1% (1/91) of the oral samples. Both the frequency and magnitude of HPV-16-specific cervical IgA was significantly elevated in women with CIN 2/3 compared with women with CIN 1 (P = 0.0073 frequency; P = 0.0045 magnitude). Women with cervical HPV-16 infection had significantly higher magnitude and frequency of cervical HPV-16 IgA responses than women without cervical HPV-16 DNA (P = 0.0002 frequency; P = 0.0052 magnitude). Despite our contention that mucosal HPV-16 antibody responses within distinct mucosal compartments may be linked, the concordance analysis carried out within and between mucosal compartments and serum suggests that no such linkage exists and that these compartments may be functioning independently of one another. An HPV-16 specific antibody response in one mucosal compartment in women with CIN is therefore not predictive of a response at another.     

Comparison of two ELISAs for the determination of Hsp70 in serum

We have compared a previously developed in-house Sandwich-ELISA with a commercial kit for the determination of heat shock protein (Hsp) 70 in serum. Samples from 64 participants were tested and there was a significant correlation between results obtained using the two assays (r = 0.807, p < 0.0001). Additionally, when ranking samples on a categorical scale, the agreement was good (72%). In the commercial test system Hsp70 was detectable in 42 (66%) of the sera, compared with 61 (95%) in the in-house ELISA method. The three samples with undetectable levels of Hsp70 in the in-house ELISA were among the 22 samples with undetectable levels of Hsp70 in the commercial ELISA kit. The apparent serum concentrations detected were different in the two systems. This dissimilarity can be ascribed to differences in the matrix used. We conclude that the in-house ELISA is more economical and performs well when measuring physiologically high, as well as low, concentrations of Hsp70.     

Comparison of mumps-IgM ELISAs in acute infection.

BACKGROUND: In Scotland there has been an outbreak of mumps with over 4000 confirmed cases in the last 2 years. As laboratory diagnosis is usually requested on patients with symptoms, a prompt diagnosis early in the illness is required. OBJECTIVES: To assess the performance of the five different commercial IgM-ELISAs used in Scottish Virus laboratories. STUDY DESIGN: The Specialist Virology Laboratory (SVC) Edinburgh distributed a serum panel to all Scottish laboratories that diagnose mumps by IgM-ELISA. The panel consisted of 45 sera from patients with confirmed mumps (date of onset known for 44/45) and 11 sera from patients with alternative diagnoses. Each laboratory performed their own commercial IgM-ELISAs blindly and reported results to the SVC Edinburgh. RESULTS: Sensitivity ranged from 24% to 51%. Assays performed better on samples taken later than 10 days after onset of symptoms. Specificity was about 82% for most assays. CONCLUSION: The sensitivity of commercial mumps IgM-ELISAs varied greatly. The Microimmune mumps-IgM ELISA had the best overall sensitivity in acute serum specimens. Diagnostic laboratories should be developing the means to perform direct detection of mumps virus for acute presentation and requesting convalescent bloods, if acute blood samples have no detectable IgM.     

人乳头瘤病毒感染检测及其临床意义

目的 探讨宫颈癌及癌前病变、尖锐湿疣与人乳头瘤病毒感染的关系,为早期防治宫颈癌提供临床依据.方法 标本来源于2004年1月至2005年8月期间重庆第三军医大学西南医院皮肤科、妇产科和解放军三○二医院皮肤科就诊的1086例患者,在未接受任何检查及治疗、无任何干预措施的情况下,采用荧光定量PCR技术对病变标本HPV-DNA进行检测;用上述方法无法明确HPV型别的标本,采用基因芯片技术检测.结果 CIN Ⅰ、CINⅡ和宫颈鳞癌的标本中,HPV的检测率均为100%,以HPV16型感染为主,但存在两种以上HPV型别感染.在子宫内膜癌中主要是HPV18,仍然存在HPV多种型别混合感染.636例女阴尖锐湿疣中,HPV阳性率为96.70%,其中以HPV6（44.97%）和HPV11（29.40%）为主,少数患者是HPV16和（或）HPV18感染,也有少数为两种以上甚至四重HPV感染.结论 子宫及宫颈瘤变主要以高危型HPV16和18为主,外阴生殖道的尖锐湿疣以低危型HPV6和HPV11为主,但两者都可合并有多种型别的HPV混合感染,给临床治疗和患者机体病理生理改变造成一定的复杂性.对HPV感染型别进行早期鉴定和监测,对提高广大妇女的生活质量有重大意义.     

Expression and control of the natural autoreactive IgG repertoire in normal human serum

We have investigated the autoreactive repertoire expressed by serum IgG of healthy individuals of various age groups using a large panel of self antigens. Natural IgG autoantibodies against all self antigens of the panel were found in the purified IgG fraction of the serum of all donors that were tested. The mean binding activity to self antigens of IgG of pregnant women was higher than that of IgG purified from the serum of infants, young adults and aged individuals. No increase in IgG autoreactivity was observed with aging neither in the purified IgG fraction of serum nor in whole serum. Whereas autoantibody activity was easily detectable in purified IgG, it was low in serum. No difference was observed, however, between the binding activity of purified IgG and of IgG in serum in the case of foreign antigens nor in the case of anti-thyroglobulin autoantibodies of patients with Hashimoto's thyroiditis. Purified IgM from normal serum bound to F(ab')₂ fragments of autologous IgG in a dose-dependent fashion and inhibited the binding of autologous IgG to self antigens. Our results thus indicate that autologous IgM contributes to regulate expression of the natural IgG autoreactive repertoire through V region-dependent interactions, resulting in low levels of IgG autoreactivity in serum under physiological conditions.     

The antigen-limited nature of microtiter ELISAs requires partial depletion of IgG to permit reliable determination of rabbit serum IgA antibody activity

The antigen-limiting nature of microtiter ELISAs predicts that antibodies of minor classes may be underestimated when the same specimen contains large amounts of IgG antibodies specific for the same antigen. Such competitive inhibition can be diagnosed from ELISA titration plots. A method is described to eliminate the negative effects of this competition on the detection of IgA antibodies in rabbit serum. The detectability of rabbit serum antibodies to ovalbumin and bovine serum albumin is increased 10-fold by prior treatment of 1:100 dilutions of serum with 1% Cowan I S. aureus. High concns of S. aureus, e.g. 10%, completely deplete serum IgG without loss of IgA. However, concns higher than 1% do not lead to additional improvement in the detectability of IgA antibodies in the systems studied. The method is rapid, inexpensive and shows no non-specific depletion of IgA from either serum or bronchoalveolar lavage fluid.     

Prevalence of infection with high-risk human papillomavirus in women in Colombia

The prevalence of human papillomavirus (HPV) infections in 2109 females inhabiting five cities of Colombia was determined. Of the 49.2% with an HPV infection, 59.8% were infected with more than one viral type. Species 7 (of the the genus Alphapapillomavirus ) was associated with multiple infections. Analysis of the socio-demographic data revealed a statistically significant protective effect associated with the status of civil union (civil recognition of cohabitation without marriage), and indigenous ethnicity proved to be a risk factor for HPV infection. This is the first study comparing HPV infection among women from geographical regions of Colombia with different socio-cultural structures.     

Prevalence and genotype distribution of cervical human papillomavirus infection: Comparison between pregnant women and non-pregnant controls

Controversies exist on the effect of pregnancy on human papillomavirus (HPV) infection. A cross-sectional section study was conducted to compare the prevalence and genotype distribution of cervical HPV infection between pregnant and non-pregnant women in Hong Kong. Cervical samples were collected from 308 pregnant women and from the same number of age-matched controls recruited from a cervical cancer screening center located at the same hospital. HPV was detected by the polymerase chain reaction, followed by genotype identification by restriction fragment length polymorphism and direct sequencing analyses. The prevalence of HPV for pregnant women was 10.1%, without significant variation with age, gestation, gravidity and parity. The prevalence of HPV for non-pregnant group was 11.4% and did not show significant difference when compared to the pregnant group either by overall or age-stratified subgroup analyses. When the analysis was stratified according to the risk-type of HPV infection, still no significant difference between pregnant and non-pregnant groups was observed (all types: 10.1 vs. 11.4%, P = 0.602; high-risk types: 5.8 vs. 7.8%, P = 0.338; low-risk types: 1.0 vs. 2.9%, P = 0.080; unknown-risk types: 3.2% vs. 1.3%, P = 0.105). The results of this study show no evidence for an influence of pregnancy on HPV prevalence, and a majority of HPV-infected pregnant women had normal cervical cytology. HPV positive results in pregnant women per se should be managed conservatively.     

Activation of the interleukin-32 pro-inflammatory pathway in response to human papillomavirus infection and over-expression of interleukin-32 controls the expression of the human papillomavirus oncogene

High-risk variants of human papillomavirus (HPV) induce cervical cancer by persistent infection, and are regarded as the principal aetiological factor in this malignancy. The pro-inflammatory cytokine interleukin-32 (IL-32) is present at substantial levels in cervical cancer tissues and in HPV-positive cervical cancer cells. In this study, we identified the mechanism by which the high-risk HPV-16 E7 oncogene induces IL-32 expression in cervical cancer cells. We used antisense transfection, over-expression, or knock-down of IL-32 to assess the effects of the HPV-16 E7 oncogene on IL-32 expression in cervical cancer cells. Cyclo-oxygenase 2 (COX-2) inhibitor treatment was conducted, and the expression levels, as well as the promoter activities, of IL-32 and COX-2 were evaluated in human HPV-positive cervical cancer cell lines. E7 antisense treatment reduced the expression levels and promoter activities of COX-2, which is constitutively expressed in HPV-infected cells. Constitutively expressed IL-32 was also inhibited by E7 antisense treatment. Moreover, IL-32 expression was blocked by the application of the selective COX-2 inhibitor, NS398, whereas COX-2 over-expression resulted in increased IL-32 levels. These results show that the high-risk variant of HPV induces IL-32 expression via E7-mediated COX-2 stimulation. However, E7 and COX-2 were down-regulated in the IL-32γ over-expressing cells and recovered by IL-32 small interfering RNA, indicating that E7 and COX-2 were feedback-inhibited by IL-32γ in cervical cancer cells.     

Comparison of PCR and hybrid capture methods for detection of human papillomavirus in injection drug-using women at high risk of human immunodeficiency virus infection.

We compared Hybrid Capture, a new technique for detection of human papillomaviruses (HPV), with a PCR assay based on L1 consensus primers. By both methods, the HPV prevalence was higher in human immunodeficiency virus (HIV)-positive women than in HIV-negative women. PCR had a higher sensitivity (0.89 versus 0.48) but lower specificity (0.43 versus 0.93) for detection of Pap smear abnormalities, compared to Hybrid Capture. The higher intensity of hybridization signal by PCR was related to higher estimates of viral load by Hybrid Capture.     

Correlation between human papillomavirus infection and histopathological diagnosis of women in Northeast Brazil

Cervical carcinoma is the fourth leading cause of death among women worldwide. Epidemiological studies claim that human papillomavirus (HPV) infection is a necessary condition for cervical cancer development. Knowledge of the geographic distribution of HPV is important in guiding the introduction of prophylactic vaccines. This study analyzed the prevalence of HPV infection in cervical samples obtained from women with abnormal cervical histopathological diagnosis in Northeast Brazil. The study included an analysis of 211 women whose diagnosis was confirmed for cervical intraepithelial neoplasia type 1 (CIN-1), cervical intraepithelial neoplasia type 2 (CIN-2), cervical intraepithelial neoplasia type 3 (CIN-3), and cancer. The identification of the HPV genotypes was based on the polymerase chain reaction–restriction fragment length polymorphism technique. A total of 42.7% of the samples showed a single HPV infection, while 57.3% showed multiple infections. The most common genotypes detected were HPV-16, HPV-18, and HPV-31. HPV-16, HPV-31, HPV-35, and HPV-18 were the most common types in CIN-1 with a single infection. HPV-16 and HPV-18 were the most often found in CIN-2 with a single infection. HPV-16, HPV-18, and HPV-31 were the most detected in CIN-3 with a single infection. HPV-16 and HPV-31 were the most frequent in cancer with a single infection. Multiple infection with HPV-16 shows a 2.7 times greater risk of CIN-3 (P = .04). Multiple infections for HPV with HPV-16 and excluding the HPV18/31 types, were associated with CIN-3 (P = .01). The results allowed the detection and genotyping of HPV types circulating in the population studied. These findings must be taken into account when devising vaccination strategies against HPV.     

Relationship between prevalent oral and cervical human papillomavirus infections in human immunodeficiency virus-positive and -negative women

Human papillomavirus (HPV) is an etiologic agent for both oropharyngeal and cervical cancers, yet little is known about the interrelationship between oral and cervical HPV infections. Therefore, we compared the prevalences and type distributions of oral and cervical HPV infections and evaluated infection concordance in a cross-sectional study within the Women's Interagency HIV Study cohort. Oral rinse and cervical-vaginal lavage samples were concurrently collected from a convenience sample of 172 human immunodeficiency virus (HIV)-positive and 86 HIV-negative women. HPV genomic DNA was detected by PGMY09/11 L1 consensus primer PCR and type specified by reverse line blot hybridization for 37 HPV types and beta-globin. Only 26 of the 35 HPV types found to infect the cervix were also found within the oral cavity, and the type distribution for oral HPV infections appeared distinct from that for cervical infections (P<0.001). Oral HPV infections were less common than cervical infections for both HIV-positive (25.2% versus 76.9%, P<0.001) and HIV-negative (9.0% versus 44.9%, P<0.001) women. Oral HPV infections were more common among women with a cervical HPV infection than those without a cervical HPV infection (25.5% versus 7.9%, P=0.002). The majority of women (207; 93.7%) did not have simultaneous oral and cervical infections by the same HPV type; however, the number of women who did (14; 6.3%) was significantly greater than would be expected by chance (P=0.0002). Therefore, the oral and cervical reservoirs for HPV infection are likely not entirely independent of one another.     

Population-based type-specific prevalence of high-risk human papillomavirus infection in middle-aged Swedish women

Human papillomavirus (HPV) DNA testing can be used to identify women at risk of the development of cervical cancer. The cost-effectiveness of HPV screening is dependent on the type-specific HPV prevalence in the general population. The present study describes the prevalence and spectrum of high-risk HPV types found in a large real-life population-based HPV screening trial undertaken entirely within the cervical screening program offered to middle-aged Swedish women. Cervical brush samples from 6,123 women aged 32-38 years were analyzed using a general HPV primer (GP5+/6+) polymerase chain reaction-enzyme immunoassay (PCR-EIA) combined with reverse dot-blot hybridization for confirmation and HPV typing by a single assay. In this study, 6.8% (95% CI 6.2-7.5) (417/6,123) were confirmed as high-risk HPV positive. Infections with 13 different high-risk HPV types were detected, of which HPV 16 was the most prevalent type (2.1%; 128/6,123), followed by HPV 31 (1.1%; 67/6,123). Any one of the HPV types 18, 33, 35, 39, 45, 51, 52, 56, 58, 59, or 66 was detected in 3.6% (223/6,123) of the women. Infection with two, three, and five types simultaneously was identified in 32, 5, and 1 women, respectively. The combination of PCR-EIA as a screening test and reverse dot-blot hybridization as a confirmatory test, was found to be readily applicable to a real-life population-based cervical screening. The type-specific HPV prevalence found support in previous modeling studies suggesting that HPV screening may be a favorable cervical screening strategy.     

Oral contraceptive use and human papillomavirus infection in women without abnormal cytological results

Both experimental and epidemiological data support the idea that oral contraceptive (OC) use may have a stimulating effect to a certain point on cervical carcinogenesis. The current investigation tries to answer the question whether OC use might have an influence on early human papillomavirus (HPV) infections. A total of 425 women without abnormal cytological results were examined colposcopically, and filter in situ hybridisation (FISH) was used to determine the presence of human papillomavirus (HPV) types 6, 11, 16 and 18. Eighty-one cervical specimens (19.1%) were found to be positive for one or more of the HPV types in FISH. HPV positivity was found to correlate with age and parity, being the highest among women under 25 and with less than two births. The use of OCs was inversely correlated with the presence of ectopy or dysplasia in this group of women. On the other hand, HPV positivity was not significantly higher among OC users than among non-users in any colposcopic group. Neither the type of pill used, nor the duration of use had any significant effect on HPV positivity. Further investigations are needed to evaluate the effects of OC use on more severe HPV- induced cervical lesions.     

Evaluation of virological procedures to detect fetal human cytomegalovirus infection: Avidity of IgG antibodies,virus detection in amniotic fluid and maternal serum

Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection and fetal damage largely due to maternal primary infection. Virological procedures which are able to detect HCMV fetal infection were evaluated. HCMV IgG antibodies were detected in 62.5% of the pregnant women and 1.47% had a primary infection. From March, 1992 to August, 1995, 29 seroconversions were observed, and in 64 other cases. HCMV IgM antibodies were detected in the first serological test. The mean IgG antibody avidity test (AI) was 31% for the 11 seroconversions tested and 74% in 32 cases where IgG and IgM HCMV antibodies were detected in the first serum. In the 29 HCMV seroconversions, 19 amniocentesis were carried out and 12 fetuses (41.4%) were infected in utero. In four amniotic fluids positive in culture and PCR, the fetus or newborns were infected and in one out of the two cordocentesis undertaken, hepatitis, anemia, and thrombocytopenia were noted. In four other cases, investigations seeking HCMV in amniotic fluid were negative whereas infants were infected at birth. Among the 64 cases with positive HCMV IgM and IgG antibodies detected in the first serological test, three fetuses were infected in utero, but no amniotic fluid was available in these cases. Amniotic fluids were studied in 39 cases, and HCMV detection by culture and PCR-hybridization was negative. HCMV DNA was detected in the maternal sera of five out of 21 pairs of seroconversions and in two cases on the first negative serum. The assay was also carried out on 50 of the 64 HCMV IgM positive sera. Two had detectable HCMV DNA. © 1996 Wiley-Liss, Inc.