T lymphocytes (CD3) may participate in the recurrence of cervical intraepithelial neoplasia grade III

Objective  Data from the literature demonstrate that the local and systemic immune responses seem to play an important role in the progression of cervical intraepithelial neoplasia (CIN). Our aim was to investigate whether recurrences among CIN III patients might be related to the presence of local lymphocytes, macrophage and enzyme iNOS. Methods  We analyzed 35 patients with CIN III who underwent conization and followed up for a minimum of 4 years. Using immunohistochemistry, the presence of T lymphocytes (CD3, CD8 and CD45RO), B lymphocytes (CD20), macrophages (CD68) and the expression of the enzyme iNOS were investigated. The quantity of marked cells is graded as: 0, absence of cells; 1, rare cells; 2, moderate number of cells; 3, many cells. For statistical purposes, we took the scores 0 and 1 to indicate weak marking and the scores 2 and 3 to indicate strong marking. Results  We found strong positive expression of CD3-positive T lymphocytes among CIN III patients with recurrence following conization (100 vs. 50% without recurrence, p = 0.02). We did not find any statistical differences in the expression of CD20, CD68, CD45RO, CD8 or iNOS. Conclusions  It is concluded that strong positive findings of CD3 T lymphocytes were related to recurrence following conization due to CIN III.     

子宫颈上皮内瘤变患者CD4＋、CD8＋T细胞表达与高危型HPV感染的关系

目的探讨不同级别宫颈上皮内瘤变（CIN）组织中CD4＋、CD8＋T细胞表达及与高危型人乳头瘤病毒（HR-HPV）的关系。方法对2005年1月至2008年8月在珠海市人民医院阴道镜下宫颈活检诊断为CIN并行宫颈锥切治疗的72例患者的资料进行分析,采用杂交捕获第二代方法（HC-Ⅱ）检测高危型HPVDNA含量;采用免疫组化检测30例CIN1、43例CIN2/3及10例正常宫颈组织中CD4＋、CD8＋T细胞的表达。结果 CIN2/3组的HR-HPV感染率显著高于CIN1组（P〈0.05）,CIN组HR-HPV感染率高于慢性宫颈炎组（P〈0.01）。慢性宫颈炎组、CIN1组和CIN2/3组CD4＋T细胞表达率分别为60.0%、43.3%和33.3%,CD8＋T细胞的表达率分别为70.0%、46.7%和31.0%;CD4＋、CD8＋T细胞在CIN1组及CIN2/3组的表达显著低于慢性宫颈炎组（P〈0.05）。结论 CD4＋、CD8＋T细胞表达下调可能促进宫颈HR-HPV感染的发生,并对宫颈癌前病变的发生、发展起一定的作用。     

MMP-1 and MMP-2 in the cervix uteri in different steps of malignant transformation--an immunohistochemical study.

OBJECTIVES: Enzymatic degradation of the extracellular matrix (ECM) represents a key element in the multistage process of tumor invasion and metastasis. This process requires extensive degradation of ECM components such as basement membrane collagen (type IV) and interstitial collagen (type I, II, III). Matrix metalloproteinase-2 (MMP-2) specifically cleaves collagen type IV, the major collagen of the basement membrane. MMP-1 digests interstitial collagen type I and III, the main collagen types of the stromal extracellular matrix. We investigated protein levels of MMP-1 and MMP-2 in different stages of malignant transformation. METHODS: Using the APAAP method we analyzed 10 normal cervical tissues, 11 cervical intraepithelial neoplasia 1 (CIN 1), 8 CIN 2 and 10 CIN 3 lesions, and 15 invasive squamous cell carcinomas. These data were compared with the HPV DNA status tested by hybrid capture II. RESULTS: Only a few isolated epithelial cells stained positively for MMP-1 and MMP-2 in normal cervical tissue and CIN 1 lesions. The CIN 2 and CIN 3 group displayed a heterogeneous distribution of MMP expression. 3 CIN 2 and 8 CIN 3 lesions showed strong MMP-2 and weak MMP-1 expression in the dysplastic epithelial cells. 5 CIN 2 and 2 CIN 3 lesions stained negatively. Invasive carcinomas showed a coexpression for MMP-1 and MMP-2 in malignant epithelial cells and peritumoral stroma cells. All MMP-2-positive cases tested positive for the HPV high-risk group. CONCLUSIONS: The expression of MMP-2 protein in preinvasive lesions of the cervix uteri and a consecutive coexpression of MMP-1 and MMP-2 in invasive cancer suggest a gradually increasing invasive potential. MMP-2 expression, when focally observed in high-grade squamous intraepithelial lesions of the cervix, may indicate tumor areas with an increased risk for invasive growth.     

Maternal lymphocyte subpopulations (CD45RA+ and CD45RO+) in preeclampsia

OBJECTIVE: The maternal syndrome of preeclampsia has been attributed to a systemic intravascular inflammatory response and endothelial cell dysfunction. The stimulus responsible for intravascular inflammation in preeclampsia has not been determined. The expression of CD45 isoforms on the surface of human T cells has been used to classify CD4(+) T lymphocytes into na?ve cells (CD45RA+) and memory T cells (CD45RO+). An increased percentage of CD45RO+ cells has been interpreted as consistent with previous exposure to microbial products or other antigens. The purpose of this study was to determine whether preeclampsia is associated with a change in the proportion of CD45RA+ and CD45RO+. STUDY DESIGN: A prospective study was conducted in patients with preeclampsia (n = 24) and normal pregnancy (n = 75). The percentage of CD45RA+ and CD45RO+ on CD4(+) T lymphocytes in peripheral blood was determined using flow cytometry and monoclonal antibodies. Results were reported as a percentage of CD4(+) lymphocytes. Parametric statistics were used for analysis. A probability value of <.05 was considered statistically significant. RESULTS: Patients with preeclampsia had a significantly higher percentage of CD45RO+ than normal pregnant women (P <.01). A significantly lower percentage of CD45RA+ was found in patients with preeclampsia than in normal pregnant women (P <.01). CONCLUSION: Preeclampsia is associated with an increase in the percentage of CD45RO+ and a decrease in the CD45RA+ lymphocyte subpopulation. Therefore, patients with preeclampsia have evidence of previous antigenic exposure, the nature of which remains to be established.     

CD8+CD45RA+CD27-CD28-T-cell subset in PBL of cervical cancer patients representing CD8+T-cells being able to recognize cervical cancer associated antigens provided by HPV 16 E7

OBJECTIVE: In response to antigenic stimulation, naive MHC-class I restricted and antigen-specific CD8+CD45RA+CD28+T-cells undergo clonal expansion and differentiate into CD8+CD45RO+ memory T-cells. Upon re- encounter with the nominal antigen, CD45RO+ T-cells are able to convert to CD8+CD45RA+CD28-T-cells displaying potent immune effector functions, including TNF-alpha production. This T-cell subpopulation constitutes a minor population in healthy individuals. In the present study we are currently evaluating whether this particular T-cell subset in PBL represents CD8+T-cells which may be able to recognize cervical cancer associated antigens provided by HPV 16 E7. MATERIAL AND METHODS: Flow-cytometric cell sorted CD8+CD45RA+CD28- and CD8+CD45RA+CD28-T-cells were obtained from patients with cervical cancer and tested for recognition of HLA-A2 restricted peptides derived from the human papillomavirus (HPV)16-E7 gene product using ELISA. HPV DNA in tumor tissue was detected by PCR. RESULTS: We show that the effector CD8+CD45RA+CD28-T-cell subset is expanded in peripheral blood lymphocytes (PBL) from patients with cervical cancer, but also in PBL from patients with an acute mycobacterial infection. CD8+T-cells from 3/6 cancer patients showed a peptide-specific immune response which could be segregated in peptide epitopes which elicited either a strong TNF-alpha production, or GM-CSF and IL-2 secretion. Peptide-reactivity could exclusively be detected in the ex vivo freshly isolated CD8+CD45RA+CD28-T-cell population. A similar situation was found to be true for HLA-A2 presented peptide epitopes derived from M. tuberculosis-associated antigens presented to T-cells obtained from patients with tuberculosis. CONCLUSIONS: The sorting of CD8+CD45RA+CD28-T-cells enables to determine the fine specificity of CD8+ effector T-cells without the need for in vitro manipulation and aids to define the most appropriate target epitopes for novel vaccine designs.     

Local immune response in squamous cell carcinoma of the uterine cervix

The role of Langerhans cells as antigen-presenting cells was examined in cervical carcinomas. Frozen samples were obtained from 34 women with stage Ib and II cervical carcinomas. Langerhans cells (CD1), T lymphocytes (CD4 and CD8), B lymphocytes (CD22), and natural killer (CD57, NK) cells were all quantitatively assessed in cervical carcinomas using immunohistochemical methods. These results were related to the MHC class I and II expression on the tumor cells. The majority of Langerhans cells were distributed among cancer cells and they were positively correlated with CD4+, NK and B cells in cervical carcinomas. This is suggestive of the presence of local immune response. The numbers of Langerhans, CD4+, CD8+ and NK cells did not significantly correlate with age at operation, lymph node metastases or depth of cervical wall invasion. The downregulation of MHC class I expression found in 8 (24%) carcinomas was not associated with the decrease in the number of immunologic cells. The upregulation of MHC class II expression found in 26 (76%) carcinomas was significantly associated with the increase in the number of Langerhans cells (p < 0.007). However, the association between the upregulation of MHC-II expression and CD4+ cells did not reach statistical significance (p < 0.07). This is probably due to a small case in this study. MHC-II-restricted immunity may partly contribute to the local immune response in stages Ib and II squamous cell carcinoma of the uterine cervix.     

Lymphocyte subset distribution and cytokine secretion in third trimester decidua in normal pregnancy and preeclampsia

BACKGROUND: Excessive Th1 activity in peripheral blood plays a probable role in the pathogenesis of preeclampsia. The aim of the study was to investigate whether disturbed local immune reactions are also present in decidua. METHODS: Flow cytometric analysis of CD3, CD19, CD56/CD16, CD4, CD8, CD4/CD29, CD4/CD45RA, CD4/CD45RO, CD8/CD28, CD3/CD69 lymphocyte subsets isolated from third trimester decidua of pregnants with preeclampsia (n=21) and pregnant controls (n=11) subjected to elective caesarean sections. Spontaneous and phytohemaglutynine stimulated "in vitro" secretion of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta by decidual lymphocytes was studied by ELISA. For the statistical significance of differences between the groups the U Mann-Whitney test was performed (confidence interval P<0.05). RESULTS: Preeclamptic patients were characterized with an increased percentage of the CD3-/CD56+CD16+, CD8+/CD28+ and decreased percentage of CD3+, CD19+, CD4+/CD45RA+ lymphocytes. The profile of secreted cytokines shifts in favor of Th1 activity (extremely high IFN-gamma and low IL-6 and IL-10 secretion). Decidual IL-12 secretion in preeclamptic patients is decreased compared to controls. CONCLUSION: Changes in NK and T lymphocyte subsets followed with Th1 cytokine IFN-gamma over-activity, could affect local immunoregulatory mechanisms in third trimester decidua of preeclamptic patients.     

Lymphatic vessel density and epithelial D2-40 immunoreactivity in pre-invasive and invasive lesions of the uterine cervix

OBJECTIVE: We sought to determine the significance of lymphatic vessel density (LVD) in pre-malignant lesions and carcinomas of the uterine cervix and to evaluate the prognostic value of lymphatic invasion and D2-40 positivity in tumor cells in the three histological types of invasive lesions. The correlation of LVD, lymphatic invasion and D2-40 positivity in tumor cells with EGFR and COX-2 expressions was also evaluated. METHODS: We studied 50 cervicitis, 50 low-grade squamous intraepithelial lesions (LSIL) (CIN1), 51 high-grade squamous intraepithelial lesions (HSIL) (CIN2/CIN3), 49 invasive squamous cells carcinomas (SCC), 43 adenocarcinomas (AC) and 30 adenosquamous cells carcinomas (ASC). The immunoreaction assay was performed using the monoclonal antibody D2-40. RESULTS: Significant differences in LVD were found among all categories of pre-invasive and invasive lesions (p=0.001 and p<0.001, respectively). LVD in invasive lesions was significantly greater than in pre-invasive lesions (p<0.001) and no significant association was found between LVD in invasive lesions and both lymph node invasion and/or metastasis. D2-40 positivity in tumor cells was associated with a better prognosis in ASC cases. EGFR and COX-2 expressions in invasive lesions were not associated with LVD; however, they correlated with both lymphatic invasion and D2-40 positivity in tumor cells. CONCLUSIONS: Lymphatic neovascularization begins early in intraepithelial lesions and continues to increase towards malignancy. Both lymphatic invasion and decrease in D2-40 expression in tumor cells appear to have a prognostic value.     

Local immune response in endometrial carcinomas

Objective To determine whether Langerhans cells act as antigen-presenting cells in endometrial carcinomas and their related lesions.
Samples Frozen endometrial samples were obtained from 13 women with normal menstrual cycles, 3 postmenopausal women, 11 women with hyperplasia (simple 4, complex 4 and atypical 3) and 32 women with endometrial carcinomas.
Main outcome measures Langerhans cells (CD1), T lymphocytes (CD4 and CD8), B lymphocytes (CD22), natural killer (NK) cells (CD57) and HLA-DR were all quantitatively assessed in endometrial samples using immunohistochemical method.
Results The numbers of Langerhans, CD4+, CD8+ and B cells were higher in the secretory phase than in the proliferative endometrium. The CD8+ cells appeared to be more plentiful than the CD4+ cells. When compared with the proliferative endometrium, the numbers of Langerhans cells were higher in hyperplasias and carcinomas. Most of Langerhans cells were HLA-DR+, showing a strong correlation with CD4+ cells in carcinomas. This suggests that MHC class II antigen restricted lymphocytes in carcinomas are activated by HLA-DR+ Langerhans cells. However, epithelial expression of HLA-DR in carcinomas did not show on association with high numbers of Langerhans and CD4+ cells. No correlation was observed between Langerhans cells and clinicopathologic features of carcinomas. In contrast, the number of NK cells significantly decreased in noninvasive carcinomas but increased in Grade 3 tumours.
Conclusion Based on the above findings, Langerhans cells are considered to act as antigen-presenting cells in carcinomas, but it was not shown that they were activated by epithelial expression of HLA-DR in carcinomas.     

DC-sign+ CD163+ macrophages expressing hyaluronan receptor LYVE-1 are located within chorion villi of the placenta

The purpose of this study was to investigate with immunohistochemical methods antigen presenting cells and their relationship to blood and lymphatic vessels in human term placenta. Fetal placental antigen presenting cells, historically also known as Hofbauer cells, were located in the chorionic villi below the syncytiotrophoblast and in the vicinity of fetal capillaries. DC-SIGN/CD209 expression was observed on CD163+, CD68+, CD45+, HLA-A,B,C+, DC-LAMP/CD208-, CD86-, Langerin/CD207-, FXIIIa-, CD1a- cells consistent with the macrophage nature of these cells. These fetal DC-SIGN+ cells lack HLA-DR, -DP, -DQ expression. Moreover, we show for the first time that they co-express the hyaluronan receptor LYVE-1. In contrast, no LYVE-1+ vessel structures, i.e. lymphatic vessels, were detected. Human term decidua hosted a variety of CD45+ cells, further phenotyped as CD163+, DC-SIGN+, CD68+, HLA-DR+, HLA-A,B,C+. Mature dendritic cells were never observed in human term placenta. In summary, human term placenta is an immunoprivileged organ without lymphatic drainage and with numerous DC-SIGN+ macrophages within the chorionic villi. We hypothesize that these cells may fulfil a function in innate responses against pathogens as well as be involved in the homeostasis of hyaluronan metabolism in the rapidly differentiating placenta.     

HIV+ patients have increased lymphocyte infiltrates in CIN lesions

OBJECTIVE: The objective of our study was to analyze immunocyte infiltrates in CIN lesions from HIV+ patients to assess whether local immunosuppression, defined as a decrease in T cell infiltrates, could explain the aggressive nature of CIN in HIV-infected patients. MATERIALS AND METHODS: Cervical tissue was obtained from 6 HIV+ CIN patients, 6 HIV- CIN patients, who underwent LLETZ (large loop excision of the transformation zone) for CIN, and 17 normal patients who underwent hysterectomy for benign indications. The following cell surface markers were analyzed: CD20 (B cells), CD4 (T helper cells), and CD8 (T suppressor/cytotoxic cells). Each tissue section was visualized with a Leica microscope at 400x and the image was captured for analysis by Harmony Group image analysis software. RESULTS: A significantly higher number of lymphocytes (both B and T cells) was detected in the stroma of HIV+/CIN tissue compared to either HIV-/CIN or normal tissue. There was also a significant increase in CD8+ cells in the HIV+/CIN group compared to HIV-/CIN or normal tissue. There was a trend toward a decreased CD4+/CD8+ ratio in the HIV+/CIN compared to the other two groups; however, this did not reach statistical significance. CONCLUSIONS: This study indicates that HIV+/CIN cervical tissue has a greater number of tissue lymphocytes recruited to the neoplastic site compared to HIV- individuals. In addition, HIV+ patients may have a decreased CD4/CD8 ratio in locally infiltrating immunocytes in CIN lesions. The local immunomodulatory effects of HIV may be detectable early in infection and therefore may explain the aggressive nature of CIN in the HIV+ patient.     

Expression of matrix metalloproteinase-9 in squamous cell carcinoma of the uterine cervix-clinicopathologic study using immunohistochemistry and mRNA in situ hybridization

OBJECTIVE: Invasion of the extracellular matrix and blood vessels by malignant neoplasms, with subsequent distant dissemination, is a key event in tumor progression. This process appears to be mediated largely through the action of matrix metalloproteinases (MMPs), a family of proteolytic enzymes produced by both stromal and tumor cells. The role of gelatinases (MMP-2 and MMP-9) in basement membrane and matrix degradation was described in various tumors. We studied MMP-9 protein expression in cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma using immunohistochemistry and detected MMP-9 mRNA using in situ hybridization. METHODS: Fifty squamous cell carcinomas, 10 cases of CIN II-III, and 10 normal cervices were stained for MMP-9, using a monoclonal antibody. The presence of MMP-9 mRNA was studied using in situ hybridization. Results were correlated with patient survival during a follow-up period of up to 167 months (average, 41 months). RESULTS: Immunohistochemical staining of tumor cells for MMP-9 was noted in 36/50 (72%) carcinomas and 5/10 (50%) CIN lesions, but was uniformly absent from the nonneoplastic epithelium adjacent to tumors and from control cervices. Peritumoral staining of stromal cells was observed in 27/50 (54%) carcinomas, but only in 3/10 (30%) CIN lesions and 1/10 (10%) control cervices. The presence of MMP-9 mRNA was detected in tumor cells in 39 (78%) carcinomas and 8 (80%) CIN lesions, but only in 4 (40%) control cervices. An intense signal for MMP-9 mRNA was observed most frequently in carcinomas. MMP-9 mRNA was detected in stromal cells in the majority of cases. However, an intense signal was observed only in stromal cells around invasive tumors. In survival analysis, age (P = 0.016), grade (P = 0. 016), and stage (P = 0.001) showed independent correlation with poor survival. Neither MMP-9 protein expression nor an intense signal for MMP-9 mRNA was associated with poor survival, although the latter was observed more frequently in neoplastic cells of lethal tumors (8/14 tumors vs 11/36 nonlethal tumors). CONCLUSIONS: MMP-9 mRNA and protein expression are elevated in tumor and stromal cells of both high-grade CIN and invasive squamous cell carcinoma of the uterine cervix. Thus, MMP-9 is possibly an early marker of tumor progression in squamous lesions of the cervix. An intense stromal signal for MMP-9 mRNA characterizes some invasive carcinomas. Expression of MMP-9 in cervical carcinoma cells is present in both lethal and nonlethal tumors, consistent with the key role of this proteolytic enzyme in invasion, and does not appear to predict disease outcome.     

Effect of IgG therapy on lymphocyte subpopulations in the peripheral blood of Kuwaiti women experiencing recurrent pregnancy loss

Intravenously administered polyspecific IgG is being increasingly used as an immunomodulating therapy with controversial beneficial outcome. The aim of this study was to evaluate the effects of IgG infusion on peripheral T-cell subpopulations in women with recurrent pregnancy loss (RPL). Fifteen women with a history of three previous RPL between 6 and 22 weeks of gestation and positivity for the antiphospholipid antibody syndrome (APS) were randomized to one of two treatment groups: (a) an intravenous immunoglobulin therapy group (RPL-IVIg; 7 patients), 500 mg IVIg/kg/month and (b) a placebo-treated group given multivitamins (8 patients). Control groups comprised either normal pregnant women without APS (10 patients) or non-pregnant women. The T-cell markers were characterized using a monoclonal antibody panel including CD3, CD4, CD8, CD25, CD29, CD38, CD45RA, CD45RO, CD54 and HLA-DR. Analysis was performed with a two-color fluorescent-activated flow cytometer. In the first trimester, the percentage of CD4+CD25+, CD4+CD45RO+, CD8+HLA-DR+, and CD8+CD38+ populations were reduced in the multivitamin group compared to normal pregnant women (p < 0.05) while in the RPL-IVIg group only CD4+CD25+ cells were reduced (p < 0.05). By the second trimester, CD3+CD16+CD56+ was significantly higher in multivitamin- than in IVIg-treated women (p < 0.05). The percentage of CD4+HLA-DR+ was significantly higher in the two RPL groups compared to normal pregnant women (p < 0.05). IVIg therapy in women with RPL was associated with a significant reduction in CD3+CD16+CD56+ and CD4+CD25+. This may contribute to the suppression of immune-mediated processes contributing to premature abortion.     

Influence of paternal lymphocyte immunization on the selected subpopulations of peripheral blood lymphocytes in women with recurrent spontaneous abortions of unknown etiology

The aim of the study was to evaluate the influence of alloimmunization on the composition of PBL subpopulations in women with recurrent spontaneous abortions (RSA) of unknown etiology. MATERIAL AND METHODS: 25 women with history of 3-6 consecutive RSA were selected for this study. Immunization with paternal lymphocytes, isolated from 100 ml of peripheral blood, was performed two times before conception (4-weeks interval). The following parameters were studied: (%) of peripheral T CD3+, T CD4+, T CD8+, T CD4+/CD45-RO+, T CD4+/CD29+, T CD8+/CD45-RO+, BCD19+ and NK CD16+/CD56+ lymphocytes; (%) of T CD3+ cells with markers of early (CD69+) and late activation (HLA-DR+) and presenting with IL-2 receptors (CD25+); the influence of alloimmunization on the expression of CD69+ receptors on cultured T CD3+ lymphocytes activated by PHA (CD3+/CD69+/FCS). RESULTS: The (%) of T CD3+ (61.0% vs. 68.6%), T CD4+ (37.4% vs. 45.3%) cells as well as T CD4+/T CD8+ ratio (1.3 vs. 1.0) significantly increased, whereas the (%) of T CD8+ (41.3% vs. 32.5%) and NK CD16+/CD56+ cells (23.9% vs. 16.2%) decreased after alloimmunization. The procedure didn't influence the (%) of T CD4+/CD45-RO+, T CD4+/CD29+, T CD8+/CD45-RO+ and BCD19+ lymphocytes. We observed significant increase of the (%) of T CD3+/CD69+ (4.4% vs. 7.3%) and T CD3+/CD25+ (5.8% vs. 10.9%) cells after immunotherapy. Moreover we observed the significant increase of the CD69+ expression on T CD3+ cells activated by PHA (CD3+/CD69+/FCS) (42.1 vs. 47.1%). CONCLUSIONS: Our study suggests that alloimmunization of women with RSA of unknown etiology results in increase of T lymphocytes' activity and a shift of the immune balance towards T helper and T supressor lymphocytes.     

Detection of human papillomavirus DNA in human cervical lesions by tissue in situ nucleic acid hybridization

Cervical cancer is one of the most common female cancers in Taiwan. Certain types of human papillomavirus (HPV) are frequently detected in the epithelial precancerous and cancerous lesions of the cervix. By the use of tissue in situ hybridization, we investigated the relationship of various types of HPV (group I, HPV-6 & 11, group II, HPV-16 & 18, group III, HPV-31, 33 & 35) with cervical condyloma, carcinoma as well as precancerous lesions. Group I HPV DNAs were mainly found in cervical condylomatous lesions (2/2) of the cervix and cervical intraepithelial neoplasia I (CIN I) (2/4), but were only occasionally found in CIN II (1/4), CIN III (1/9) or non-keratinized squamous cell carcinoma (1/15). HPV DNAs of groups II and III were mainly detected in lesions of CIN III (5/9) and invasive squamous cell carcinoma (large cell, keratinized type: 4/7; large cell, non-keratinized type: 11/15). HPV DNA sequences were invariably detectable only in the cell nuclei of condyloma or dysplastic epithelium or invasive carcinoma. However, they could not only be detected in the upper layer dysplastic cells and koilocytes but also in the well and poorly differentiated cervical cancer cells. The distribution of HPV DNA positive cells in the carcinomas fell into four different patterns: (1) upper zone and non-invasive regions of the carcinoma (11/22, 50%), (2) basal zone and invasive regions (2/22, 9%), (3) randomly scattered (7/22, 32%), and (4) extensively distributed over the whole tumor lesions (2/22, 9%). Thus, our results are consistent with a strong correlation between the presence of HPV-16, 18, 31, 33 and 35 and malignant conversion of cervical epithelial cells.     

妊娠期糖尿病患者母儿免疫水平变化的临床研究

目的:研究妊娠期糖尿病(GDM)患者母儿免疫球蛋白(Ig)、补体(C)、T淋巴细胞亚群以及NK细胞水平的变化,探讨GDM对母儿体液免疫和细胞免疫的影响。方法:选择58例不同糖耐量水平的GDM患者为研究对象根据是否需要胰岛素治疗进一步分为GDM1组和GDM2组,以30例健康孕妇做对照。免疫比浊法测定外周静脉血与脐血的免疫球蛋白、补体水平;流式细胞技术测定外周静脉血与脐血的T细胞亚群及NK细胞水平。结果:GDM组外周血CD4+、CD4+/CD8+、CD16+CD56+、IgG含量均下降,CD8+、C3、C4、IgE含量升高,差异有统计意义(P<0.05);CD3+、IgA、IgM含量下降,差异无统计学意义(P>0.05)。GDM组脐血中CD3+、CD4+、CD4+/CD8+、CD16+CD56+含量均下降,差异有统计学意义(P<0.05);IgM、IgG含量下降,CD8+、IgE含量升高,差异无统计学意义(P>0.05)。与GDM1组相比,GDM2的指标呈现更明显的变化趋势。结论:妊娠期糖尿病存在母儿体液免疫和细胞免疫的失衡,且病情越重,这种改变越显著。     

Squamous carcinoma of the uterine cervix with CIN 3-like growth pattern: An under-diagnosed lesion

Abstract. Al-Nafussi AI, Monaghan H. Squamous carcinoma of the uterine cervix with CIN 3-like growth pattern: An under-diagnosed lesion.
Invasive squamous carcinomas of the cervix have traditionally been classified into keratinizing, non-keratinizing, verrucous, warty (condylomatous), papillary transitional (squamo-transitional), and lymphoepithelioma-like carcinomas. The majority of these tumors are easily recognized. We present for the first time the pathological appearances of six cases of invasive squamous carcinoma with growth pattern simulating tangentially cut CIN 3 involving endocervical glandular crypts/clefts. In all cases initial diagnosis on biopsy and/or loop excision was thought to be CIN 3, perhaps with suspicion of early invasion. On further excision and/or on clinical grounds the tumors were frankly invasive. We propose the use of the term squamous carcinoma with "CIN 3-like growth pattern" for such lesions. This is in order to avoid misinterpretation as CIN 3 with subsequent inappropriate management of patients with this type of tumor.     

Alterations in the expression of selected MHC antigens in premalignant lesions and squamous carcinomas of the uterine cervix

BACKGROUND: The objective of the study was to determine whether there is a correlation between expression of selected major histocompatibility complex (MHC) antigens and the presence of carcinoma and premalignant lesions of the cervix. We also attempted to determine whether there is a correlation between expression of the selected MHC antigens in each grade of cervical intraepithelial neoplasia (CIN) and invasive carcinoma of the cervix. METHODS: Quantitative comparative analysis of MHC class I and class II expression between specimens of the uterine cervix was carried out. For detecting human leukocyte antigen (HLA) class I molecules, two primary antibodies were used, recognizing either epitopes of all free heavy chain class I molecules or epitopes of the HLA-Bw4 antigen. HLA class II antigens were detected by antibodies reacting with the alpha-chain of the HLA-DR molecule. An indirect immunoperoxidase technique was carried out using paraffin-embedded sections. The following groups of sections were formed: squamous metaplasia, CIN of different grades, and invasive carcinoma. The specimens were taken from women in whom routine histopathologic diagnosis of cervical lesions had been performed. RESULTS: A significant decrease in the mean number of stained cells showing expression of MHC class I antigens (HC 10 and Bw4) was found in the invasive cancer group of specimens. A decrease in Bw4 expression was also found in the CIN III group. The highest expression of the HLA-DR antigens was detected in the metaplastic epithelium compared to the other groups. In the dysplastic epithelium the expression of the HLA-DR antigens was increased in the highest grade of dysplasia (CIN III). Upregulation of HLA-DR expression was detected in invasive cancer specimens. CONCLUSIONS: Alterations in MHC expression are found in premalignant lesions and squamous carcinomas of the uterine cervix. Quantitative assessment with the use of an image analysis system is valuable in helping to discriminate staining patterns of HLA expression. Further studies are indicated to evaluate the prognostic value of the expression of selected MHC antigens.     

CD1a和E-cadherin在宫颈上皮内瘤变中的表达

目的:探讨CD1a和E-cadherin与宫颈癌发生发展的关系及作为早期癌变生物学指标的可能性。方法:采用免疫组化SP法检测CD1a和E-cadherin在同期56例宫颈上皮内瘤变、56例宫颈鳞癌和15例正常宫颈组织中的分布及表达。结果:(1)CD1a+朗格汉斯细胞在正常宫颈、CIN和宫颈癌各组中数量逐渐减少,两两比较有显著差异(P0.01);CIN中该细胞数量随病变严重程度减少,CINⅠ和CINⅡ、CINⅠ和CINⅢ组之间两两比较有显著差异(P0.05),而CINⅡ与CINⅢ组间无显著差异(P0.05);(2)E-cadher-in在正常宫颈、CIN和宫颈癌各组中的阳性表达率及强度逐渐下降,两两比较有显著差异(P0.05);在CIN中阳性表达率及表达强度随病变严重程度呈下降趋势,CINⅠ和CINⅡ、CINⅠ和CINⅢ之间两两比较有显著差异(P0.05),而CINⅡ和CINⅢ之间无显著差异(P0.05);(3)宫颈组织中CD1a+朗格汉斯细胞的细胞数与E-cadherin的阳性表达率及强度呈正相关(r=0.912,P0.05)。结论:CD1a+朗格汉斯细胞与E-cadherin可能在宫颈癌的发生、发展过程中起重要作用。     

Decidual lymphocyte subsets in pregnant women

OBJECTIVES: Materno-foetal immunological reactions in decidua are probably one of the most important elements in pathogenesis of preeclampsia. DESIGN: To compare lymphocyte subsets isolated from decidua of preeclamptic pregnant women with lymphocyte subsets isolated from healthy pregnant women. MATERIALS AND METHODS: Preeclampsia (PE) was defined according to USA National Health Institute criteria. The study group consisted of 21 women with PE and 11 women with physiological pregnancy. All pregnancies were finished with elective cesarean sections. Exclusion criteria were: uterine contractions, infection, chorinamnionitis, diabetes mellitus and therapy with steroids less than 7 days before blood sampling. Decidual tissue was obtained by curettage of the uterine cavity. The fragments of decidua were separated from clotted blood and placed in sterile tubes with 5 ml of isotonic solution (PBS). Then the decidual tissue was mechanically fragmented, homogenized and rinsed in PBS. Routine immunofluorescent marking techniques with monoclonal antibodies were performed. Analysis was done with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The following lymphocyte subsets were estimated: CD3, CD19, CD4, CD8, CD4/CD29, CD8/CD28, CD4/CD45RA, CD4/CD45RO, CD56/CD16, CD69. The results were described as percentage of lymphocytes positive for above surface molecules. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118 15 grant. RESULTS: Decidua of pregnant PE women contains significantly increased percentage of CD3-/ CD56 + 16+, CD8+/CD28+ cells and decreased percentage of CD3+, CD19+, CD4+/CD29+ and CD4+/CD45RA+ compared to decidua of healthy pregnant controls. CONCLUSIONS: These changes suggest that deficiency of suppressor activity as well as aberrant immunoregulation exists in decidual tissue of PE women.