Human papillomavirus type influences the extent of chromosomal lag during mitosis in cervical intraepithelial neoplasia grade III.

The level of risk for carcinoma in the uterine cervix depends on the type of human papillomavirus (HPV) present. We examined whether the HPV type influences the proliferation rate and occurrence of mitotic figures with lagging chromosomes in the precursor of cervical carcinoma. The study group comprised 180 women who were referred because of cytologic changes indicating dysplasia and who were subsequently diagnosed with cervical intraepithelial neoplasia grade III. The HPV-16-associated lesions showed a significantly higher number of mitoses per 1,000 nuclei than the lesions without HPV (p < 0.001). The HPV-16-associated lesions also showed a significantly higher proportion of mitotic figures with lagging chromosomes than did the HPV-18- or HPV-31-associated lesions and lesions without HPV (p = 0.01, p = 0.007, and p = 0.002, respectively). Our results indicate that the differences in oncogenic potential among HPVs become apparent in the precursor lesion through the differences in the extent of chromosomal lag during mitosis.

The suitability of DNA cytometry for the prediction of the histological diagnosis in women with abnormal cervical smears.

OBJECTIVE: To analyse the suitability of DNA cytometry for predicting the histological diagnosis in women with cervical dyskaryosis. DESIGN: Survey with the use of diagnostic information to revise disease probability. SETTING: Colposcopy clinic of a university hospital. PARTICIPANTS: One hundred and ten women with two mildly or moderately dyskaryotic cervical smears and 98 women with one severely dyskaryotic smear. INTERVENTIONS: DNA cytometric analysis using cytocentrifuge preparations of single cell suspensions from a cervical scrape. The main DNA cytometric parameter was N5C (i.e. the absolute number of cells with a DNA content of more than 5C on a given surface with a predefined cell density). MAIN OUTCOME MEASURE: The probability of finding CIN II or worse. On arbitrary grounds, a positive test should point to a probability of 85% or higher. RESULTS: In the patients with cervical neoplasia, the value of N5C increased significantly with an increasing CIN grade (P < 0.001). In the patients with one severely dyskaryotic smear and in those with two mildly or moderately dyskaryotic smears, the prior probability of finding CIN II or worse was 94% and 53%, respectively. Therefore, DNA cytometric analysis might be particularly useful in women with mild or moderate dyskaryosis; further analysis was restricted to this group. All of the women in whom the N5C value was higher than 52 were diagnosed as having CIN II or worse. Only 16 (14.5%) of the 110 women had an N5C value of 52 or higher. When the N5C value was 27, the probability of finding CIN II or worse was estimated to be 85%. Only 28 (25%) patients had an N5C value of 27 or higher. CONCLUSIONS: DNA cytometry produced significant diagnostic information, as was shown by the relation between N5C and the histological diagnosis. However, the N5C value could not discriminate sufficiently between women with CIN II or worse and CIN I or better. Therefore, the management of individual patients with cytological abnormalities cannot be based on the results of DNA cytometric analysis.

Epidemiological evidence of cervical intraepithelial neoplasia without the presence of human papillomavirus.

The aim of this paper was to provide epidemiological evidence to support the notion that cervical intraepithelial neoplasia (CIN) without human papillomavirus (HPV) is a true entity. If a diagnosis of HPV-negative cervical neoplasia is erroneous, one would not expect there to be any differences in risk factors between HPV-positive and HPV-negative patients. Patients at a gynaecological outpatient clinic of a university hospital [a total of 265 consecutive women with dyskaryotic cervical smears who were subsequently diagnosed with CIN I (n=37), CIN II (n=48) or CIN III (n=180)] completed a structured questionnaire regarding smoking habits and sexual history. Analysis of an endocervical swab for Chlamydia trachomatis, analysis of a cervical scrape for HPV, and morphological examination of cervical biopsy specimens were also performed. HPV was found in 205 (77.4%) out of the 265 women. Univariate analysis showed that current age (P=0.02), current smoking behaviour (P=0.002) and the number of sexual partners (P=0.02) were significantly associated with the presence of HPV. Age at first sexual intercourse, a past history of venereal disease or genital warts, and current infection with Chlamydia trachomatis were not associated with the presence of HPV. Using multivariate logistic regression analysis, the number of sexual partners and current smoking behaviour showed an independent significant association with HPV. HPV-negative and HPV-positive CIN patients differ with respect to the risk factors for HPV. These findings suggest that HPV-negative CIN is a separate true entity.

Bacterial vaginosis is not important in the etiology of cervical neoplasia: a survey on women with dyskaryotic smears.

BACKGROUND AND OBJECTIVES: It has been suggested that bacterial vaginosis may play a role in the etiology of cervical neoplasia. The authors analyzed the prevalence, risk factors, and impact on histologic changes of bacterial vaginosis in women with cytological abnormalities of the uterine cervix. METHODS: Two-hundred-eighty women with dyskaryotic smears were surveyed. Using a questionnaire, data were obtained on smoking habits and sexual history. Bacterial vaginosis was the diagnosis if the vaginal discharge produced a fishy odor upon alkalinization and if clue cells were seen in the wet smear. Cervical scrapes were analyzed for the presence of human papillomavirus DNA, and cervical tissue specimens were analyzed for the presence and severity of (intraepithelial) neoplasia and the proliferation rate (mitotic index) of the lesion. Chlamydia trachomatis was identified by culture of an endocervical swab. RESULTS: Bacterial vaginosis was found in 56 (20%) out of the 280 women. The presence of bacterial vaginosis was significantly associated with the number of cigarettes smoked per day, age at first sexual intercourse, the lifetime number of sexual partners, and current Chlamydia trachomatis infection. The number of cigarettes currently smoked per day and the lifetime number of sexual partners were independent significant risk factors for the presence of bacterial vaginosis. There was no relation between the presence of bacterial vaginosis and the human papillomavirus infection. Bacterial vaginosis did not influence the severity of the (intraepithelial) neoplasia or the mitotic index. CONCLUSION: In women with dyskaryotic cervical smears, the prevalence of bacterial vaginosis did not seem to be increased, and bacterial vaginosis did not influence the histologic changes. Therefore, bacterial vaginosis is unlikely to be important in the etiology of cervical neoplasia, despite the similarity between its epidemiologic features and those of cervical human papillomavirus infection and cervical neoplasia.

Atypical mitotic figures and the mitotic index in cervical intraepithelial neoplasia.

We surveyed cervical intraepithelial neoplasia (CIN) to quantify the proliferation rate and the presence of normal and atypical mitotic figures. In the cervical tissue specimens of 127 women with CIN, the area with the highest cell proliferation was identified and, at that site, the proliferation rate was assessed by calculating the mitotic index (MI). Lesions with an MI < 2 were not considered further. In the area with the highest proliferation rate, 228 mitoses were classified into one of the following groups: normal mitotic figures (NMFs), lag-type mitoses (OLTMs) comprising three group metaphases (3GMs), two group metaphases (2GMs) and other lag-type mitoses (LTMs), multipolar mitoses (MPMs) comprising tripolar mitoses (3PMs) and quadripolar mitoses (4PMs), and other atypical mitotic figures (OAMFs). The median value of the MI increased significantly from 3 in CIN I through 4 in CIN II to 9 in CIN III (P < 0.001). The occurrence of the different LTMs was mutually correlated. The frequency of LTMs increased significantly with increasing CIN grade (P < 0.001), whereas the frequency of NMFs decreased significantly with increasing CIN grade (P < 0.001). The frequency of OAMFs was not related to CIN grade (P = 0.94). MPMs were present in low numbers in a minority of the lesions. Spearman's rank correlation coefficient (with 95% confidence limits) between the MI and the number of LTMs, OAMFs and NMFs was 0.66 (0.53; 0.75), -0.14 (-0.32; 0.05) and -0.51 (-0.63; -0.35), respectively. Increasing CIN grade is associated with increasing MI, increasing numbers of LTMs, and decreasing numbers of NMFs. MPMs are very rare events in CIN. The abundant presence of OAMFs seems to be independent of CIN grade and MI.

Cigarette smoking and human papillomavirus in patients with reported cervical cytological abnormality.

OBJECTIVE: To assess the relation between two risk factors for cervical neoplasia: smoking and infection with oncogenic human papillomavirus. It has been suggested that smoking causes a local immunological defect, which could facilitate the infection and persistence of human papillomavirus. DESIGN: Cross sectional epidemiological study. Completion of a structured questionnaire by the patients, analysis of cervical scrapes for human papillomavirus, and morphological examination of biopsy specimens. SETTING: Outpatient gynaecological clinic. SUBJECTS: 181 women with a report of cervical cytological abnormality. MAIN OUTCOME MEASURES: Prevalence of infection with oncogenic human papillomavirus and smoking habits. RESULTS: Oncogenic human papillomavirus was found in the cervix of 26 (41%) of the 63 women who did not smoke, 22 (58%) of the 38 who smoked 1-10 cigarettes a day, 28 (61%) of the 46 who smoked 11-20 cigarettes a day, and 26 (76%) of the 34 who smoked > or = 21 cigarettes a day. The prevalence of the virus thus increased in accordance with the number of cigarettes smoked (p = 0.001). This relation remained after adjustment for age at first intercourse and lifetime number of sexual partners. Of the 63 non-smokers, 23 had previously smoked at least 10 cigarettes a day at some time. Of these 23 women, 14 (61%) had oncogenic human papillomavirus in their cervix. Of the 40 women who had never smoked at least 10 cigarettes a day, 12 (30%) had the virus. The prevalence of oncogenic human papillomavirus in non-smokers therefore depended on previous smoking habits (p = 0.03). CONCLUSION: The dose dependent effect of cigarette smoking on the occurrence of oncogenic human papillomavirus favours a causal relation between these risk factors for cervical neoplasia.

Three-group metaphase as a morphologic criterion of progressive cervical intraepithelial neoplasia.

OBJECTIVE: The purpose of our study was to investigate the presence of three-group metaphase in progressive cervical intraepithelial neoplasia. STUDY DESIGN: This was a retrospective histologic study on the conization specimens of 41 women with microinvasive cervical carcinoma, 28 of whom were enrolled in the study. Three-group metaphase was scored in the invasive part of the lesion and in the adjacent cervical intraepithelial neoplasia. RESULTS: Three-group metaphase was found in 93% of cervical intraepithelial neoplasia adjacent to the invasive part of the lesion. However, three-group metaphase was found in 11% of the microinvasive cervical carcinoma cases with an infiltration depth of less than 2.5 mm and in 60% of the microinvasive cervical carcinoma cases with an infiltration depth of between 2.5 and 5.0 mm. CONCLUSION: The chance of finding three-group metaphase seems to be limited by the area of the lesion examined for three-group metaphase on the slide. Given the relation between three-group metaphase and aneuploid cervical intraepithelial neoplasia found in the literature and the occurrence of three-group metaphase in the cervical intraepithelial neoplasia adjacent to the microinvasive cervical carcinoma in this study, three-group metaphase can be considered a morphologic criterion for progressive cervical intraepithelial neoplasia and can be of value for practical use.

The three-group metaphase as a morphologic indicator of high-ploidy cells in cervical intraepithelial neoplasia.

There is a need for additional morphologic criteria to improve the value of histologic classification for the prediction of the biologic behavior of cervical intraepithelial neoplasia (CIN). Representative slides from 72 cone specimens containing CIN were examined to study the correlation between the presence of three group metaphases (TGMs), a morphologically well defined and light microscopically readily recognizable atypical mitotic figure, and the incidence of aneuploid cells with a nuclear DNA content greater than 5C. The numbers of cells greater than 5C (minus the polyploid cells 8C +/- 1C) were counted, using LEYTAS image cytometry on Cytospin preparations from the 72 blocks corresponding to the slides searched for TGMs and used for histologic classification of the lesions in classes CIN 1-3. It appeared that large numbers of aneuploid cells greater than 5C were more closely related to the presence of TGM than to a higher CIN class per se, particularly in women older than 35. Since aneuploid CIN has a higher progression rate than euploid CIN, the presence of TGMs will indicate a biologically unfavorable lesion. Thus, TGM deserves further investigation as an additional morphologic parameter for predicting the biologic behavior of CIN.

Human papillomavirus and the three group metaphase figure as markers of an increased risk for the development of cervical carcinoma.

In this study, the presence of atypical mitotic figures and human papilloma virus (HPV) genomes was related to the degree of cervical intraepithelial neoplasia (CIN) or microinvasive carcinoma (MIC) as found in 94 paraffin-embedded biopsies from cervical lesions. The results showed that the frequency of three group metaphase (TGM) figures, a special kind of atypical mitotic figure, as well as the presence of HPV 16 and 18 genomes increased with the degree of cervical intraepithelial neoplasia. TGM figures were observed in 24% of CIN2, up to 61% in CIN3 lesions, and in 83% of the microinvasive cervical carcinomas. HPV genomes were detected in 15% of CIN1, up to 75% in CIN3 lesions, and in 92% of the invasive carcinomas of the cervix. The combination of these two markers showed even a better association with a higher degree of cervical intraepithelial neoplasia. The results of these studies suggest that detection of particular HPV types, mainly HPV 16 and 18, and the presence of TGM figures can be considered as markers that indicate an increased risk for progression of cervical intraepithelial neoplasia to invasive carcinoma.