子宫颈病变组织中低分子量蛋白酶体表达与HPV16感染的关系

目的观察低分子量蛋白酶体(low molecular-weight protein,LMP)在子宫颈病变组织中的mRNA和蛋白表达,探讨其与HPV16感染的关系。方法以152例新疆维吾尔族妇女正常子宫颈上皮、子宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)和子宫颈鳞癌(cervical squamous cell carcinoma,CSCC)患者为研究对象,采用RT-PCR和免疫组化法鉴定LMP2和LMP7mRNA及蛋白表达水平;采用PCR技术检测相应标本HPV16感染情况。结果 (1)LMP2、LMP7随着子宫颈病变的加重其蛋白表达逐渐降低,且mRNA表达水平与蛋白表达趋势相一致。在CIN中LMP2、LMP7蛋白表达下调和缺失率分别为25.0%/15.6%、29.7%/23.4%;在子宫颈癌中LMP2、LMP7蛋白的表达下调和缺失率分别为17.5%/34.9%、23.8%/41.3%。临床病理参数进行分析发现LMP2、LMP7与子宫颈癌分化程度及淋巴结转移密切相关(P<0.05)。(2)PCR结果显示,HPV16的检出率随着子宫颈病变的进展而增加,在慢性子宫颈炎、CIN和子宫颈癌组织中阳性率分别为8%(2/25)、67.2%(43/64)和77.8%(49/63),且随着肿瘤恶性程度的增加其阳性表达率增加,各组间阳性表达差异有统计学意义(P<0.05)。在CIN中LMP7表达下调与HPV16感染有关(P<0.05),子宫颈癌中LMP2和LMP7表达下调与HPV16感染有关(P<0.05)。结论 LMP基因的转录表达下调或蛋白质表达缺失与维吾尔族妇女子宫颈癌病变进程密切相关,其中HPV16感染可能是重要原因之一。     

Papanicolaou's nebular cells are related to HPV infection and HSIL

Metaplastic cells with nebular cytoplasmic changes in the cervical smear are classified in the Dutch coding system for cervical screening as KOPAC O8 cells. Since these nebulated cells are already documented by Papanicolaou, we refer to these cells as Papanicolaou's nebular cells. We examined the simultaneous presence of these characteristic metaplastic cells and high-grade squamous intraepithelial lesion (HSIL) in a population-based data base from January 1991 and December 1996. The odds ratio (OR) of nebular cells concurring with HSIL increases with age. For the age cohort 30 years, the OR was 7.8 with a 95% confidence interval (CI) of 4.4-13.9. For the age cohort 60 years, the OR was 35.3 with a 95% CI of 7.8-159.2. Aiming to determine the nature of these nebular metaplastic cells, we performed Chlamydia and HPV PCR on 587 and 1,483 smears, respectively. With an OR of 0.9 [0.3-2.4] it is unlikely that Chlamydia plays a role in the appearance of these nebular cells in the smear. This study shows that with an OR of 5.9 [1.7-21.3] HPV is not only related to large koilocytosis but also to a nebular change of small metaplastic cells. This study reports that nebular changes of small metaplastic cells are related to cervical cancer and to HPV infection.     

Performance of the Abbott RealTime high‐risk HPV test in women with abnormal cervical cytology smears

HPV DNA testing is known to be much more sensitive than cytology, but less specific. A range of HPV and related tests in 858 women referred for colposcopy because of an abnormal smear were evaluated to compare the performances of these tests. This article compared the Abbott test to other tests which had been previously evaluated. This test was a real true test for 14 high‐risk HPV types. The Abbott test was found to be highly sensitive for cervical intraepithelial neoplasia grade 3 or worse (CIN3+) (98.9%) with a specificity of 31.5%. These numbers were comparable with the Qiagen HC2 test, the Roche Linear Array and Amplicor tests, and the Gen‐Probe APTIMA test. Differences between these tests appeared to be related mostly to the choice of cutoff level. An added feature of the Abbott test was the provision of type specific results for HPV 16 and 18. J. Med. Virol. 82: 1186–1191, 2010. © 2010 Wiley‐Liss, Inc.     

Morphometric evaluation and nonclassical criteria for the diagnosis of HPV infection and cytological atypia in cervical samples

Herein, we evaluated cervical samples from normal tissue or HPV‐infected tissue, to determine if the relative nuclear/cytoplasmic ratio (NA/CA) and the presence of nonclassical cytological criteria are a novel cytological criterion for the diagnosis of HPV. Significantly, larger NA/CA ratios were found for the HPV?ATYPIA+ and HPV+ATYPIA+ groups compared with HPV?ATYPIA? group, regardless of collection method. For the samples collected with a spatula, only three samples from the HPV?ATIPIA? group showed four or more nonclassical parameters (i.e., were positive), while a larger number of the samples in the HPV?ATYPIA+, HPV+ATYPIA?, and HPV+ATYPIA+ groups were positive (13, 4, and 13 samples, respectively). Among those collected with a brush, no sample showed four or more nonclassical criteria in the HPV?ATYPIA? group, while a number of samples were positive in the HPV?ATYPIA+, HPV+ATYPIA?, and HPV+ATYPIA+ groups (4, 3, and 4 samples, respectively). HPV infection was associated with significant morphometrical changes; no increase in the NA/CA ratio was found in the HPV+ATYPIA‐ samples, compared with the HPV?ATIPIA? samples collected with either a spatula or a brush. In conclusion, by including nonclassical cytological criteria into the patient diagnosis, we were able to reduce the number of false negative and false positive HPV diagnoses made using conventional cytology alone. Diagn. Cytopathol. 2013;41:785–792. © 2013 Wiley Periodicals, Inc.     

Dutch solutions for liquid-based cytology: analysis of unsatisfactory slides and HPV testing of equivocal cytology

The liquid-based techniques to obtain microscopy slides for cervical screening have replaced conventional smears almost completely in the USA, but not in all European countries. The decision making process to use liquid-based cytology (LBC) for nationwide screening programs depends on the health system. In a pilot study of over 7,000 screenees, we analyzed the unsatisfactory LBC slides and tested the equivocal cytologies for HPV by using the LiPA test. For comparison over 48,000 conventional screening data were used. Compared to conventional smears, the LBC slides were highly cellular, the state of fixation was much better, and obscuring blood did not exist. The unsatisfactory rate showed an increase from 262/100,000 (conventional smears) to 357/100,000 (LBC slides) due to too thick, undiagnosable epithelial fragments on the LBC slides. HPV testing of the equivocal cytology leads to a better patient management and less unnecessary referrals.     

高危亚型HPV33和HPV52感染与宫颈病变的关系

目的 探讨人乳头瘤病毒(HPV)高危亚型HPV33和HPV52感染与宫颈病变的关系。方法 选取316例于我院进行病理组织活检联合HPVDNA检测的病例,对其进行HPVDNA提取、PCR扩增、导流杂交,其中对194例HPV33和HPV52亚型阳性患者资料进行分析。结果 194例HPV33和HPV52阳性患者中,宫颈良性病变225例,宫颈上皮内瘤变(CIN)为114例和宫颈恶性肿瘤为55例,利用统计学χ~2检验发现在HPV33和HPV52阳性患者中,CIN和宫颈恶性肿瘤的阳性率明显高于宫颈良性病变的阳性率(p<0.05)。结论 HPV33和HPV52阳性患者患宫颈病变的几率增大,高危型HPV33和HPV52是引起宫颈恶性病变的致病基因型。     

Distribution of human papillomavirus among women with abnormal cervical cytology in Kuwait

This study investigates the distribution of human papillomavirus (HPV) in women with abnormal cervical cytology in Kuwait. Two hundred and ninety‐eight (298) abnormal ThinPreps were taken from women seeking routine gynecological care and screened for HPV DNA by real‐time PCR. HPV genotyping was determined by PCR‐based sequencing. HPV DNA was detected in 152 women (51%), and 29 different HPV genotypes were detected, comprising 16 high‐risk (HR) (16, 18, 31, 33, 35, 39, 45, 51, 53, 56, 58, 59, 66, 68, 73, 97), nine low‐risk (LR) (6, 11, 54, 61, 74, 81, 90, 102, 106), and four intermediate‐risk (IR) (62, 67, 84, 87). HPV16 had the highest prevalence (24.3%), followed by HPV11 (13.8%), HPV66 (11.2%), HPV33 (9.9%), HPV53 (9.2%), HPV81 (9.2%), HPV56 (7.9%) and HPV18 (6.6%). HPV prevalence was 86, 67, and 89% in women with invasive cervical carcinoma (ICC), high‐grade squamous intraepithelial lesion (HSIL) and low‐grade squamous intraepithelial lesion (LSIL), respectively. As for age distribution, 69% of all HPVs were found in women aged 20–29 years, and the HPV incidence rate deceased with increasing age. The proportion of single infections decreased as the severity of the cytological diagnosis increased, while the proportion of multiple infections increased. This study is the first of its type in Kuwait and one of few in the Middle East. The findings are consistent with the hypothesis that HPV infection is the primary cause of cervical neoplasia. They support HPV vaccine research to prevent cervical cancer and efforts to develop HPV DNA diagnostic tests. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Prevalence of human papillomavirus among women with normal cervical cytology in Kuwait

This study was undertaken to determine the prevalence and type specific distribution of human papillomavirus (HPV) in women with normal cervical cytology in Kuwait. The study is the first of its type in Kuwait and one of few in the Middle East. The age specific distribution of HPV types was determined in 3,011 ThinPrep samples taken from women seeking routine gynaecological care. ThinPrep samples were screened for HPV DNA by real-time PCR. The type specific distribution of the viruses was determined by PCR-based sequencing. The results showed that HPV DNA was detected in 71 women (2.4%), and 21 different HPV genotypes were detected, comprising eight high-risk (HR) (16, 31, 33, 53, 56, 58, 66, and 73), seven low-risk (LR) (6, 11, 54, 61, 70, 81, and 90), four intermediate-risk (IR) (67, 82, 83, and 84) and HPV 102 and HPV 106. LR HPV types were found in 71.8% of infected samples, HR types in 32.3%, and IR types in 7%. With regard to age, 40.8% of all HPVs were found in women 30-39 years of age, 29.6% in women 40-49 years of age, 19.7% in women over 50 years and 9.9% in women less than 34 years old. The study shows that the prevalence of HPV infection in Kuwait is among the lowest in the world and suggests that HPV vaccine could prevent the development of HPV associated cervical cancer in 1.39% of young females living in Kuwait. However, more extensive population-based studies should be undertaken before implementing HPV vaccination.     

Association between human papillomavirus infection and cytological abnormalities during early follow-up of invasive cervical cancer

Considering the limitations of cytology for detection of residual cervical cancer after radiotherapy, the aim of this study was to evaluate the frequency and viral load of high-risk HPV in cervical and vaginal samples during the early follow-up of patients treated for invasive cervical cancer and correlate the results with cytological examination. Conventional cytology and hybrid capture test were performed on cervical and vaginal samples of 52 women with invasive cervical carcinoma 3 months after therapy. High-risk HPV was detected in 46.1% of the samples and the median and the range of the ratio relative light unit (RLU)/cutoff (CO) (estimated viral load) in positive samples was 1.71 (1-2120.03). No significant difference was observed in viral frequency and in median of the ratio RLU/CO between samples of patients at different stages (I-III) and between cervical samples, from patients treated by exclusive radiotherapy with or without chemotherapy, and vaginal samples, from patients who underwent to hysterectomy and radiotherapy with or without chemotherapy. Cytological abnormalities were more frequent significantly in samples with HPV than in samples without HPV. The viral load was also higher significantly in samples with cytological abnormalities when compared with the samples without cytological abnormalities. In conclusion, HPV detection methods may be useful during the early follow-up as a complement to conventional cytology for the diagnosis of residual cervical cancer after radiotherapy.     

Accuracy comparison between PAPNET diagnoses and conventional diagnoses in an Italian cervical cytology laboratory

In a population-based cervical screening laboratory with a controlled workload, a retrospective PAPNET review of 1,654 Pap smears was performed. The series included a random sample of smears originally and conventionally classified as negative (no. 1,309) and unsatisfactory (no. 43), and 299 consecutive smears originally classified as abnormal. The PAPNET diagnoses and the original diagnoses were compared for accuracy. For smears with discordant classification, a majority diagnosis as obtained on manual review was used as a gold standard. The PAPNET testing showed a greater proportion of negative smears classified as unsatisfactory (3.7% vs. 0.2%; ratio, 16.7; 95% CI 12.4–22.0); a reduced proportion of ASCUS/LGSIL cases correctly identified as positive (i.e., a reduced sensitivity for such lesions) (76.3% vs. 97.9%; ratio, 0.78; 95% CI 0.66–0.91); an equal proportion (96.2%) of AGUS/HGSIL cases correctly identified as positive (i.e., an equal sensitivity for such lesions); and an equal proportion of negative smears correctly identified as such (i.e., an equal specificity). In conclusion, the PAPNET testing did not improve the diagnostic performance of the manual screening. Diagn. Cytopathol. 1998;19:279–283. © 1998 Wiley-Liss, Inc.     

Role of HPV DNA,HPV mRNA and cytology in the follow‐up of women treated for cervical dysplasia

The aim of this study was to assess the role of cytology, human papilloma virus (HPV) DNA and human papilloma virus messenger RNA (HPV mRNA) assays in detecting cervical intraepithelial neoplasia grade 2+ (CNi 2+) (recurrences/persistence) during the follow‐up of women after treatment of cervical intraepithelial lesion. This cross‐sectional study was performed among 43 women treated for cervical intraepithelial neoplasia (CIN) between January 2014 and January 2017 at the Department of Obstetrics and Gynecology of Spedali Civili's Hospital, Brescia, Italy. Pap smear and cervical samples for HPV tests were collected during the follow‐up visit. Furthermore, colposcopy was always performed in order to find out the persistence/recurrence of the disease. A cervical biopsy was collected when necessary. Cervical samples obtained were tested for HPV DNA using the INNO‐LiPa HPV assay and for HPV mRNA using the APTIMA assay. The mean age of enrolled women was 42.5 years. Among the treated patients, more than 50% of women revealed the absence of high risk HPV DNA and HPV mRNA. We found the persistence of the disease cervical intraepithelial neoplasia grade 2 (CIN 2) only in one woman. The sensitivity of cytology, HPV DNA and HPV mRNA in detecting disease was satisfactory (100%), while the specificity was quite different for the three tests: 64.2, 52.4 and 78.9%, respectively. The HPV mRNA test has higher specificity with respect to cytology and HPV DNA, avoiding the referral to unnecessary colposcopy with an improvement of costs/benefits for healthcare system. However, given the small size sample, this study should be considered as a pilot for future larger studies.     

Resolving ASCUS without recourse to HPV testing: manual reprocessing of residual automated liquid-based cytology (ALBC) material using manual liquid-based cytology (MLBC)

We show that residual cell material from ThinPrep PapTest (Cytyc Corporation, Boxborough, MA) atypical squamous-cells of undetermined significance (ASCUS) cases may be manually reprocessed to triage women into actionable diagnostic categories (HSIL, LSIL, and Negative). Material remaining from each of 358 ThinPrep ASCUS cases was manually reprocessed as two slides, labeled "A" and "B." Interobserver agreement between case contributors (CCs) and three sequential reviewers (SRs) was analyzed with 186 cases (Study 1), and diagnostic reproducibility between SRs was examined with an additional 172 cases (Study 2). In Study 1, CCs classified 33% of cases as LSIL or greater, SRs classified 60% as LSIL or greater, and there was 58% diagnostic agreement between CCs and SRs. No "Negative" case assignment by one group afforded an "HSIL" assignment by the complementary group. In Study 2, there was 95% agreement between SRs A slide and B slide diagnoses with 54% of A slides and 55% of B slides classified as LISL or greater. Again, no "Negative" case assignment to one slide afforded an "HSIL" assignment to the complementary slide. Overall, 12.6% of the 358 cases showed HSIL, and all HSILs by one observer group were ASCUS or greater by the complementary observer group. Using manual reprocessing beyond the 21-day specimen outdate for HPV testing by the Hybrid Capture II High Risk HPV test (HR-HCII; Digene Corporation, Beltsville, MD), many ThinPrep ASCUS cases were reclassified as LSIL or HSIL. The 12.6% HSIL proportion of this study approximated the 11.5% CIN 2 or greater proportion of the ALTS ASCUS arm. Similar to ALTS, manual liquid-based cytology (MLBC) would have referred nearly 60% of women to colposcopy for a definitive diagnosis of HSIL or LSIL without resorting to HPV testing. These data demonstrate that many cases of automated liquid-based cytology (ALBC)-diagnosed ASCUS represent unrecognized SIL, which is present in the ALBC specimen vial at the time the ASCUS diagnosis is rendered.