Functional variants in CYP1A1 and GSTM1 are associated with clearance of cervical HPV infection.

OBJECTIVE: We evaluated time to clearance of high risk (HR) HPV infection in relation to functional variants in three genes (CYP1A1, GSTT1, and GSTM1). METHODS: The study group consisted of 450 HR-HPV infected women from the Atypical squamous cells of undetermined significance-low-grade squamous intraepithelial Lesion Triage Study (ALTS) cohort followed up at the clinical center at Birmingham, Alabama. The Cox proportional hazard model with the Wei-Lin-Weisfeld (WLW) approach was used, controlling for relevant covariates. RESULTS: Women who were polymorphic for CYP1A1 experienced an HR-HPV clearance rate that was 20% (HR=0.80, p=0.04) lower than women without the polymorphism for CYP1A1, adjusting for all other cofactors. The GSTM1 null genotype was associated with higher HR-HPV clearance rate (HR=1.39, p=0.006). The polymorphism in GSTT1 was not significantly associated with time to clearance of HR-HPV. CONCLUSIONS: Xenobiotic metabolism genes may influence the natural history of HR-HPV infection and its progression to cervical cancer.

Genomic amplification of the human telomerase RNA gene for differential diagnosis of cervical disorders.

To evaluate genomic amplification of the human telomerase RNA gene (TERC) as a supportive approach to cytopathology or histopathology in diagnosis of low-grade and high-grade uterine cervical lesions, 1,033 Chinese women at three medical centers had liquid-based thin-layer cytopathologic examination and TERC detection by fluorescence in situ hybridization (FISH). Human papillomavirus DNA testing, colposcopy with or without biopsy, and histopathologic examination were conducted as needed. In cytopathologic examination, genomic amplification of TERC was found in 30 of 659 (4.6%) normal or benign cellular changes; in 23 of 170 (13.5%) atypical squamous cells of undetermined significance (ASCUS); in 8 of 28 (28.6%) atypical squamous cells with high-grade squamous intraepithelial lesion possible (ASC-H); and in 26 of 103 (25.2%) low-grade (LSIL) and 64 of 73 (87.7%) high-grade (HSIL) squamous intraepithelial lesions; with pairwise significant difference (P< 0.05) in each, except ASC-H and LSIL (chi(2) = 0.127, P = 0.72). In histopathologic examination, TERC was amplified in 28 of 671 (4.2%) normal, inflammatory, or wart cases; in 17 of 233 (7.3%) cervical intraepithelial neoplasia grade 1 cases (CIN 1); in 27 of 39 (69.2%) CIN 2 cases; in 57 of 67 (85.1%) CIN 3 cases; and in 22 of 23 (95.7%) cervical cancer cases; with pairwise significant difference in each (P < 0.05). The number of cells with abnormal signals increased and the abnormal signal patterns were diversified with increasing severity of cervical dysplasia. FISH detection of TERC amplification may provide an effective, noninvasive approach in conjunction with cytopathologic or histopathologic evaluation for differential diagnosis of low- and high-grade cervical disorders.

Evaluation of two types of sponges used to collect cervical secretions and assessment of antibody extraction protocols for recovery of neutralizing anti-human papillomavirus type 16 antibodies.

Immunogenicity evaluations in human papillomavirus (HPV) vaccine trials have relied on serological samples, yet cervical antibodies are likely to be most relevant for protection against infection. In order to assess functional antibody levels at the cervix, the secreted-alkaline-phosphatase neutralization assay (SEAPNA) was used to measure HPV-neutralizing activity. We assessed the variability of the SEAPNA with serum samples after vaccination with an HPV type 16 (HPV16) L1 virus-like particle vaccine and whether the SEAPNA can be used to monitor neutralizing activity at the cervix. The SEAPNA has an overall coefficient of variation of 29.3%. Recovery from ophthalmic sponges was assessed by spiking V5 (mouse anti-HPV16) antibody onto and extracting it from sterile Merocel and Ultracell sponges and sponges used to collect specimens from participants. V5 recovery from sterile Merocel sponges was complete, yet that from Ultracell sponges was null. The mean V5 recoveries from participant Ultracell and Merocel sponges were 61.2% and 93.5%, respectively, suggesting that Merocel sponges are more appropriate for specimen collection. The SEAPNA can be applied to determine the surrogates of protection and to examine the durability of protection at the cervix.

Telomerase activity and human papillomavirus in malignant, premalignant and benign cervical lesions.

The purpose of this study was to define a correlation between telomerase activity and human papillomavirus (HPV) in normal control tissue and in benign, premalignant and malignant cervical lesions. Telomerase activity was detectable in 33 out of 34 cases of squamous-cell carcinoma, five out of six cases of microinvasive carcinoma, 8 out of 20 cases and two out of six cases of high- and low-grade squamous intraepithelial lesions (SILs) respectively. The higher frequency of positive telomerase in invasive carcinoma compared with SILs was observed in both HPV-associated and non-associated groups. Whereas 92.6% of HPV-positive and 100% of HPV-negative invasive lesions expressed telomerase, only 50% of HPV-positive and 25% of HPV-negative SILs did. Interestingly, telomerase activity was also detectable in 13 out of 28 cases of benign lesions regardless of the presence of HPV. In conclusion, there may be two roles of telomerase in the cervix. The first one would present in benign lesions; the second is associated with cancer development and activated during the late stage of multistep carcinogenesis in both HPV-positive and -negative groups.

Association of cervicovaginal infections with increased vaginal fluid phospholipase A2 activity.

OBJECTIVE: The purpose of this study was to determine if phospholipase A2 was detectable within vaginal fluid and to correlate its presence with the presence of common lower genital tract infection or microbial conditions. STUDY DESIGN: Pregnant women were examined at the first prenatal visit with standard clinical evaluations and microbiologic cultures or tests. Vaginal fluid samples were evaluated for phospholipase A2 activity by means of a standardized enzyme fluorometric assay. Data were stratified to control for coexisting infections. RESULTS: Phospholipase A2 activity was detected among 29.8% of women and was independently associated with the presence of bacterial vaginosis (p < 0.001), Trichomonas. vaginalis (p < 0.04), and Chlamydia trachomatis (p < 0.02). The percentage of women with phospholipase A2 activity and the level of activity was increased in the presence of more than one infection. CONCLUSIONS: Elevated reproductive tract phospholipase A2 concentrations among pregnant women may play roles in the pathogenesis of preterm labor and birth. Identification of pregnant women with increased concentrations in vaginal fluid may allow for development of effective intervention strategies to reduce the risk of preterm birth.

Placental form of glutathione S-transferase in normal and diseased human uterine cervical mucosa.

Expression of the human placental form of glutathione S-transferase (GST-pi), an enzyme proposed as a marker for human and experimental neoplasia, was immunohistochemically evaluated in 51 samples of 'normal' and diseased adult human uterine cervix. Five fetal uteri were also studied. GST-pi positivity was detected in 54, 92, 95 and 83% of the 'normal', non-neoplastic, cervical intra-epithelial neoplasia (CIN) and cancer cases respectively. All five fetal uteri and the positive 'normal' adult cases presented cells immunostained for GST-pi throughout the thickness of the mucosa, including the basal layer. Some non-neoplastic conditions like inflammation, repair and metaplasia and some dysplastic and neoplastic lesions showed areas of positively stained cells within an otherwise negative tissue, indicating a phenotypic heterogeneity regarding the enzyme expression. Our results confirm that GST-pi has a fetal character and indicate that it may appear in the adult cervical squamous epithelia under 'normal' or pathological conditions not necessarily linked to the process of carcinogenesis. Therefore it cannot be used as a marker for cervical epithelial neoplasia.

Histoenzymological study on desquamative cervico-vaginal cells.

Dihydrofolate dehydrogenase activity was histochemically investigated comparatively to the activities of 6-phosphogluconate, glucose-6-phosphate, malate and lactate dehydrogenases, NADH-2-tetrazolium dehydrogenase, esterases, beta-glucuronidase and leucyl aminopeptidase in smears of desquamative cervico-vaginal cells of negative and positive Papanicolaou classes. Dihydrofolate dehydrogenase was found active in a greater percentage of cells and stronger in individual cells of cervical carcinoma than in normal, inflammatory or dysplastic cervix uteri. The other enzymes activities were less specifically increased in dyskaryotic cells.

Histochemical studies of glycogen metabolizing enzymes in normal and abnormal human cervical epithelium.

The stratified squamous epithelium in eleven cervical cone biopsies was studied for localization of glycogen, glycogen synthetase, branching enzyme and amylophosphorylase in areas showing normal, dysplastic epithelium and carcinoma in situ. In the normal cervix glycogen was found to be present in the intermediate and superficial cells, amylophosphorylase activity was present in the basal and parabasal layers and the branching enzyme was found largely in the lower and middle zone of the intermediate layer. As the degree of abnormality increased, a progressive decrease of glycogen and branching enzyme and increasing amylophosphorylase was found, suggesting that amylophosphorylase could be a constitutive enzyme in normal, dysplastic and neoplastic cells. The method for localization of glycogen synthetase activity was not sufficiently reproducible nor sensitive for conclusions to be drawn.