[Show abstract][Hide abstract] ABSTRACT: To delineate the potential role of p21-activated kinases (PAKs) in the pathogenesis of gestational trophoblastic diseases (GTD) by defining the expression pattern of PAK-1, -4 and -6 and their potential implication in estrogen receptor (ER) regulation of normal placental tissue and GTD.
We evaluated immunohistochemically 10 normal first-trimester placentas (NP), 10 partial moles (PM), 15 complete moles (CM) and 3 choriocarcinomas (CCA) for PAK-1, PAK-4, PAK-6 and ER expression intensity and localization. Staining outcomes were assessed utilizing non-parametric Kruskal Wallis one-way analysis of variance test followed by pairwise Wilcoxon Rank Sum tests. Statistical significance was determined by two-sided p-value of<0.05.
In NP, PAK-6 immunoreactivity was predominantly cytoplasmic. Compared to NP, PM and CM demonstrated significant increase of cytoplasmic PAK-6 in cytotrophoblast (p=0.012, p=0.033 respectively), accompanied by significantly increased nuclear immunoreactivity in cytotrophoblast (p=0.008, p=0.045 respectively) and intermediate trophoblast (p=0.003, p=0.015 respectively). PAK-4 was found significantly upregulated in both cytoplasmic and nuclear compartments of cytotrophoblast and syncytiotrophoblast in PM (p=0.004 and p=0.002 for cytotrophoblast; p=0.018 and p=0.002 for syncytiotrophoblast, respectively) and CM (p=0.001 and p=0.001 for cytotrophoblast; p=0.002 and p=0.001 for syncytiotrophoblast, respectively) when compared to NP, whereas PAK-1 expression was significantly reduced in the syncytiotrophoblast of PM (p=0.025 for cytoplasm and p=0.008 for nucleus). Nuclear expression of ER was undetectable in all stained samples.
Our results reveal PAK-6 upregulation in GTD compared to NP. The absence of nuclear expression of ER might stem in part from the repressive effect of PAK-6 in trophoblastic tissue.
No preview · Article · Sep 2013 · Gynecologic Oncology
[Show abstract][Hide abstract] ABSTRACT: To study the expression of vascular endothelial growth factors (VEGFs), placental growth factor (PLGF) and their receptors (VEGFR-1, -2, -3) and their regulators (IL-6, CD147) in normal placenta and gestational trophoblastic disease (GTD) in order to evaluate their potential role in the biology of GTD.
Paraffin sections of 10 normal, first-trimester placentas, 10 partial moles, 10 complete moles, 5 choriocarcinomas and 5 placental site trophoblastic tumors (PSTTs) were studied immunohistochemically for expression of VEGFR-1, VEGFR-2, VEGFR-3, IL-6, PLGF and CD147. Immunolocalization of VEGF, Angiopoietin-1 and Angiopoietin-2 was performed on 5 choriocarcinomas and 5 PSTTs. The levels of VEGF and VEGFR-2 were determined in supernatants and lysates of normal trophoblast, JEG-3 and JAR choriocarcinoma cells with electrochemiluminescence assays.
The normal placenta had significantly stronger expression of VEGFR-2 than did those of partial and complete mole (p = 0.001, p = 0.003). VEGF, Angiopoietin-1 and Angiopoietin-2 expression in PSTT were significantly higher than those in choriocarcinoma (p = 0.002, p= 0.01, p = 0.038). Choriocarcinoma showed stronger intensity of staining for VEGFR-3 than did normal placenta, partial and complete mole (p = 0.036, p = 0.038, p = 0.05). Choriocarcinoma had significantly stronger staining of CD147 than did partial and complete mole (p<0.01, p<0.01). PSTT exhibited significantly stronger staining for IL-6 than did choriocarcinoma (p = 0.03).
PSTTs exhibited strong staining for VEGF, and choriocarcinoma showed strong staining for VEGFR-3. Agents that inhibit the activity of VEGF and VEGF receptors may prove to be useful in the therapy of gestational trophoblastic neoplasia.
No preview · Article · Jun 2012 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: This study aimed to investigate the expression of recently identified matrix metalloproteinases (MMPs), their inhibitors (TIMPs), and inducer (CD147) in a wide range of gestational trophoblastic diseases (GTD) thereby expanding our understanding of the potential role of MMPs in GTD.
Paraffin sections of 10 normal first-trimester placentas (NP), 10 partial moles (PM), 10 complete moles (CM), 5 choriocarcinomas (CCA) and 5 placental site trophoblastic tumors (PSTT) were studied immunohistochemically for expression of MMP-7, MMP-14, MMP-21, MMP-28, TIMP-3, TIMP-4 and CD147. Immunolocalization of MMP-1, MMP-2, MMP-3, MMP-9, MMP-13 and TIMP-1 was performed on 5 CCA and 5 PSTTs.
CCA showed stronger intensity for MMP-14 and MMP-28 than PSTT (p<0.05, p<0.05). CCA and PSTT had stronger expression of MMP-21 than NP, PM and CM (p<0.05, p<0.05, p<0.01). PSTT (p<0.05, p<0.05), NP (p<0.01, p<0.01) and CM (p<0.01, p<0.05) showed stronger staining for TIMP-3 and TIMP-4 than CCA.
Choriocarcinoma's high expression of MMPs and low expression of MMP inhibitors may contribute to its invasiveness and metastatic potential. Similarly, PSTT's lower expression of MMPs and high expression of MMP inhibitors may partly explain its lower invasiveness. Agents that inhibit MMP may prove useful in treating GTD.
No preview · Article · Jul 2011 · Gynecologic Oncology
[Show abstract][Hide abstract] ABSTRACT: Advanced maternal age may result in a weaker immune response against complete molar pregnancy, therefore increasing the risk of gestational trophoblastic neoplasia due to ineffective elimination of the trophoblastic cells after evacuation. The present study was undertaken to investigate the cellular immune response against complete molar pregnancy at the implantation site in younger and older patients.
Immunolocalization of CD8, granzyme B (GrB), FoxP3 and CD56 was performed on histologic tissue sections prepared from 18 patients aged < or = 40 years and 10 patients aged > 40 years to characterize effector (GrB+CD8+) cytotoxic T cells, GrB positive and negative natural killer cells (CD56) and regulatory T cells (FoxP3+) at the implantation site in complete molar pregnancies.
The number of the different immune cell types did not show significant differences in the implantation sites of complete molar pregnancies between the 2 age groups or between persistent and nonpersistent cases.
Immunosenescence of the natural killer and T cells most likely does not play a role in the increased incidence of gestational trophoblastic neoplasia in older patients with complete moles.
No preview · Article · May 2010 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: To review the clinical experience in the treatment of patients with low-risk gestational trophoblastic neoplasia (GTN) over the past 30 years in a national trophoblastic disease center.
Between January 1, 1977, and December 31, 2007, 302 patients with low-risk GTN were treated. The patients were directed to our institution from all parts of Hungary. The patients were 14 to 53 years of age with an average age of 28.3 years. Methotrexate (MTX)/folinic acid or actinomycin-D (Act-D) primary chemotherapy was selected based upon the patient's stage and prognostic score of GTN.
Among 218 low-risk patients, 210 (96.3%) achieved remission as a result of MTX therapy. In 8 patients (3.7%), MTX-Act-D-cyclophosphamide (MAC) combination chemotherapy was needed to achieve complete remission, in some cases assisted by operation. Among 84 patients, 81 (96.4%) achieved remission as a result of Act-D therapy. In 3 cases (3.6%) complete remission was achieved by MAC combination chemotherapy. We detected metastases in 22.8% (69/302) of our low-risk patients. Chemotherapy, surgical intervention or other supplementary treatments resulted in 100% remission in cases of low-risk nonmetastatic and metastatic disease.
Our data indicate that MTX/folinic acid or Act-D should be the primary treatment in patients with nonmetastatic or metastatic low-risk GTN. Importantly, patients with resistance to single-agent chemotherapy regularly achieve complete remission with MAC combination chemotherapy. Results show that patient care under the direction of experienced clinicians serves to optimize the opportunity for cure and minimize morbidity.
No preview · Article · May 2010 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: In general, gynecologic diseases are diseases involving the female reproductive tract. These diseases include benign and malignant tumors, pregnancy-related diseases, infection, and endocrine diseases. Among them, malignant tumor is the most common cause of death. In recent years, the etiology of some of these diseases has been revealed. For example, Human Papilloma Virus (HPV) infection has been shown to be one of the major etiological factors associated with cervical cancer. Inactivation of tumor suppressor gene BRCA1 has been implicated in hereditary ovarian cancer. In spite of these findings, the molecular bases of most of the diseases remain largely unknown. This chapter focuses on discussing benign and malignant tumors of female reproductive organs as well as pregnancy-related diseases, which have relatively well-understood molecular bases. Infections of the genital mucosa with HPVs represent the most common virus-associated sexually transmitted disease and at the age of 50 years, approximately 80% of all females will have acquired a genital HPV infection sometime during their life. The chapter presents a review of mucosal high-risk HPVs and their contributions to cervical lesions and cancers.
[Show abstract][Hide abstract] ABSTRACT: To evaluate the possible mechanisms influencing the infiltration of CD8 T lymphocytes into the tumor epithelium of advanced-stage serous ovarian cancers.
Immunohistochemical localization of CD8 T lymphocytes was done on a homogeneous population of 184 high-grade, advanced-stage serous ovarian cancer tissue specimens. Microarray analysis was done on microdissected tumor epithelium from 38 specimens to identify genes up-regulated or down-regulated in specimens with differing numbers of tumor-infiltrating CD8 T lymphocytes. Quantitative real-time PCR and immunohistochemistry were used to validate a candidate gene. Univariate and multivariate survival analyses were done combining CD8 T lymphocyte number and HLA-DMB expression with standard prognostic factors.
Marked CD8 T lymphocyte infiltration of the tumor epithelium is associated with a 20-month improvement in median overall survival. Additionally, when combined with cytoreduction status and age, CD8 T lymphocyte status is an independent prognostic factor for survival. Microarray analysis showed HLA-DMB, a component of the MHC II antigen presentation machinery, to be differentially expressed in specimens with an abundance of tumor-infiltrating CD8 T lymphocytes. This relationship was validated at both mRNA and protein levels. As well, high HLA-DMB expression in the tumor epithelium was associated with a significant improvement in median overall survival in both univariate and multivariate analyses.
Tumor cell expression of HLA-DMB is associated with increased numbers of tumor-infiltrating CD8 T lymphocytes and both are associated with improved survival in advanced-stage serous ovarian cancer.
Full-text · Article · Jan 2009 · Clinical Cancer Research
[Show abstract][Hide abstract] ABSTRACT: To determine the level of infiltration and antigen profile of immune cells and explore their relationship in intraplacental and postmolar choriocarcinoma to better understand the immunobiology of choriocarcinoma.
Immunolocalization of CD8, Granzyme B (GrB) and FoxP3 was performed on sections prepared from 5 intraplacental and 7 postmolar choriocarcinomas to characterize effector Tc cells (GrB+, CD8+) and GrB- (GrB-, CD8+) Tc cells, GrB+ NK cells (GrB+, CD8-) and Treg (FoxP3+) cells.
In the case of intraplacental choriocarcinoma, immune cell infiltration was not detected in the surrounding villi or in the tumor. Immune cell infiltration into the implantation site of the placenta with intraplacental choriocarcinoma did not differ from that into the normal pregnancy implantation site. In postmolar choriocarcinoma the immune cell infiltration of the adjacent tissue was vigorous and involved all types of immune cells (Tc, NK, Treg). In 6 of 7 cases of postmolar choriocarcinoma, in spite of the vigorous immune response, immune cells could not be seen in the choriocarcinoma tissue. A sharp infiltration border of immune cells was noted at the edge of the postmolar choriocarcinoma tissue.
Intraplacental choriocarcinoma is not associated with a vigorous immune cell response. In contrast, postmolar choriocarcinoma is associated with a vigorous immune cell response in adjacent tissues but not the choriocarcinoma tissue itself.
No preview · Article · Sep 2008 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: To determine the microvessel density (MVD) at the implantation site of normal placenta (NP) and molar pregnancies and to correlate MVD with clinical data and underlying angiogenic factors.
Immunolocalization of CD31, vascular endothelial growth factor and angiopoietin 1 and 2 were performed on NPs, nonpersistent partial moles, persistent partial moles (PPM), nonpersistent complete moles and persistent complete moles (PCM).
Significant differences were identified in the MVD between NP and complete mole (CM), and PM and CM (p < 0.001 and p < 0.035, respectively). MVD in PPM and PCM was significantly higher (p = 0.036 and p < 0.001, respectively) when compared to NP. MVD > 100 per high-power field was associated with an increased risk of persistence (p < 0.04). MVD showed a strong correlation with immediate postevacuation hCG levels (p < 0.03). Angiopoietin 2 staining was more heterogeneous, with lower overall expression in molar pregnancies as compared to more homogeneous expression in NP (p < 0.05).
MVD is highly correlated with hCG levels, suggesting that hCG may act as an angiogenic factor during implantation of molar pregnancy. MVD at the implantation site may be associated with excessive trophoblastic proliferation or reflect high hCG levels, which places patients at increased risk of persistent neoplasia.
No preview · Article · Aug 2008 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: To investigate the antigenicity of normal placenta and complete molar trophoblastic tissue.
T cell receptor (TCR) variable beta chain (VBC) gene expression was analyzed utilizing fresh frozen tissues. Real-time polymerase chain reactions (RT-PCR) were performed on cDNA samples from 10 normal buffy coats (BC), 7 normal placentas (NP) and 14 complete molar pregnancies (CM) using a TCR beta chain and 25 variable TCR beta chain primers. Relative expressions were calculated for each individual gene.
Significant changes were noted in most of the gene expressions in NP and CM as compared to the buffy coat samples. The relative expression of most genes was significantly decreased in NP and CM, but VBC gene number 4 was increased in both NP and CM; however, a significant difference was noted only between BC and CM (p = 0.023). Comparing NP to CM, 5 other VBC gene expressions were decreased significantly in the CM tissues (p < 0.05).
In normal placenta and CM pregnancies, T cells appear to express certain TCR VBC genes in a different manner than in BC. These genes and the VBC gene profile difference found between NP and CM may play an important role in the immunobiology of CM pregnancy.
No preview · Article · Aug 2008 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: Laminin receptor 1 (LR1), a non-integrin-type laminin receptor, has been described in several tumors and may play a role in tumor invasion. It is well known that LR1 can modify the conformation and degradation of laminin thus enhancing tumor cell invasion. LR1 may play a role in controlling trophoblast invasion in normal and molar pregnancies.
Real-time polymerase chain reaction (RT-PCR) was performed on total cDNA from 17 gestational age-controlled normal placentas, 10 complete moles (CM), and 4 partial moles (PM). Immunolocalization of LR1 was performed on paraffin sections prepared from 17 age-controlled placentas, 17 PM, and 19 CM.
RT-PCR demonstrated a 13-fold increase in LR1 mRNA expression in molar tissues (PM and CM combined) versus normal placentas (p=0.012). Immunohistochemical analysis revealed LR1 localized to the decidual cells. In normal placenta LR1 localized to the decidual cell membrane. However, in PM and CM, LR1 was additionally noted in the cytoplasm of decidual cells. Interestingly, with immunohistochemistry method, we found that PM demonstrated higher protein expression of LR1 than either normal placenta or CM (p=0.001 and p=0.024, respectively).
LR1 is expressed in both the decidual cells of normal placenta and mole (CM and PM). Decidual cells in CM and PM express LR1 significantly greater than the decidual cells in placenta. The underlying mechanism of how molar tissue may be associated with enhanced expression of LR1 in the maternal endometrium is unclear.
No preview · Article · Feb 2008 · Gynecologic Oncology
[Show abstract][Hide abstract] ABSTRACT: Cytotoxic T cells (Tc) and natural killer (NK) cells may play a role in controlling trophoblast invasion. This study was undertaken to determine the level of infiltration and antigen profile of immune cells and explore their relationship in normal placenta (NP) and molar tissues to understand better the biology of gestational trophoblastic diseases. Immunolocalization of CD8, Granzyme B (GrB), and FoxP3 was performed on sections prepared from 11 gestational age-matched NP, 19 partial moles (PM), and 18 complete moles (CM) to characterize effector (GrB+CD8+) and GrB- (GrB-CD8+) Tc cells, GrB+ NK cells (GrB+CD8-), and Treg (FoxP3+) cells. Immune cells infiltrated into the implantation site of normal placenta, PM, and CM with increasing frequency. Effector and GrB- Tc, GrB+ NK and Treg infiltration in the CM were significantly stronger than seen in the normal placenta (p=0.002, p=0.007, p=0.002, respectively). Immune cell infiltration was not detected in the villi or trophoblast of gestational tissues. Treg infiltration at the implantation site was only observed in PM and CM. In CM and PM Tc infiltration positively correlated with Treg infiltration (p=0.035), but Treg infiltration did not correlate with the Tc effector ratio (effector Tc cells / all Tc cells). In CM the cellular immune response at the implantation site was significantly more vigorous than seen in case of the normal placenta. These observations demonstrate that at the implantation site of CM, the number of effector Tc and GrB+ NK cells are increased and Treg cells may negatively regulate T lymphocyte activation.
[Show abstract][Hide abstract] ABSTRACT: Cytotoxic T cells (Tc) and natural killer (NK) cells may play a role in controlling trophoblast invasion. This study was undertaken to determine the level of infiltration and antigen profile of immune cells and explore their relationship in normal placenta (NP) and molar tissues to better understand the biology of gestational trophoblastic diseases.
Immunolocalization of CD8, Granzyme B (GrB), and FoxP3 was performed on sections prepared from 11 gestational age-matched NP, 19 partial moles (PM), and 18 complete moles (CM) to characterize effector (GrB+CD8+) and GrB- (GrB-CD8+) Tc cells, GrB+ NK cells (GrB+CD8-), and Treg (FoxP3+) cells.
Immune cells infiltrated into the implantation site of normal placenta, PM, and CM with increasing frequency. Effector and GrB- Tc, GrB+ NK and Treg infiltration in the CM were significantly stronger than seen in the normal placenta (p=0.002, p=0.007, p=0.002, respectively). Immune cell infiltration was not detected in the villi or trophoblast of gestational tissues. Treg infiltration in the implantation site was only observed in PM and CM. In CM and PM Tc infiltration positively correlated with Treg infiltration (p=0.035), but Treg infiltration did not correlate with the Tc effector ratio (effector Tc cells/all Tc cells).
In CM the cellular immune response in the implantation site was significantly more vigorous than seen in normal placenta. These observations demonstrate that in the implantation site of CM, the number of effector Tc and GrB+ NK cells are increased and Treg cells may negatively regulate T lymphocyte activation.
No preview · Article · Dec 2007 · Gynecologic Oncology
[Show abstract][Hide abstract] ABSTRACT: The authors review their clinical experience with the treatment of gestational trophoblastic neoplasia (GTN) over the past 25 years at the Hungarian National Trophoblastic Disease Center. Between 1st January, 1977 and 31st December, 2001 the authors treated 355 patients with GTN referred from all parts of Hungary. The patients' age varied between 14-53 years, the average being 28.3 years. The patients were selected for primary chemotherapy based upon the anatomical staging and prognostic score factors of the GTD (gestational trophoblastic disease). Out of 173 patients remission was achieved in 162 (93.2%) as a result of methotrexate therapy. In case of 11 patients (6.8%) they could obtain a complete remission by using a combination chemotherapy, in some cases assisted by operation. 5 of these 11 patients have had high-risk disease. In case of another 68 patients remission could be achieved in 63 (92.6%) as a result of Actinomycin-D therapy, whereas in 5 cases (7.4%) by a secondary combination chemotherapy. Two out of these 5 patients had a high risk disease. Chemotherapy, surgical intervention or other supplementary treatments resulted in a 100% successful therapy in cases of non-metastatic and low-risk metastatic disease in patients with GTD 95.9% curring rate (in 116 of 121 patients) was achieved in group of patients with high risk GTD using combined chemotherapy (MAC, EMA-CO, CEB) as primary treatment 98.6% total remission rate (in 350 of 355 patients) was achieved in patients with GTD at authors' center over the past 25 years. Data support the concept that patient care under the guidance of experienced clinicians serves to optimize the opportunity for cure and minimize morbidity. According to the experiences the middle-dose methotrexate or Actinomycin-D therapy should be selected as primary treatment in patients with non-metastatic or low-risk metastatic GTD. It is of importance to note that in cases of patients with a disease resistant to single agent chemotherapy, complete remission can regularly be achieved by using a combined chemotherapy. Necessary to emphasize that combined chemotherapy must be selected for primary treatment of patients with high risk GTD.
[Show abstract][Hide abstract] ABSTRACT: Recurrent miscarriage and IVF implantation failure affect millions of women annually. It is one of the great failings of the medical community that in as many as 75% of these cases cause is never established. Considering that habitual abortions may have several causes including anatomical, morphological, genetic, internal bacteriological as well as immunological abnormalities it is inevitable to pursue these kinds of medical examinations and tests. One part of the immunological background can be explained by auto-immune diseases while the other part of it is associated with immunopathological factors. Here we refer to the currently available practical testing batteries that can directly assess immunological background furthermore, we call attention to their importance. Presently there is no really accepted widely approved treatment for recurrent spontaneous abortion (RSA) or repeated IVF failure. Unlike so many other areas in medicine, the development of new treatments for recurrent miscarriage has not at all kept place with scientific advances in immunology. This will become apparent in the ensuing discussion of some of the current approaches to treating recurrent miscarriage. Here both the so called conventional therapies and the recently developed mainly immunology based treatments will be descripted and evaluated.
[Show abstract][Hide abstract] ABSTRACT: Gestational trophoblastic diseases are interrelated conditions characterized by abnormal growth of chorionic tissues with varying propensities for local invasion and metastases. These diseases are characterized by altered expression of several growth regulatory factors and oncogenes. On the basis of the expression of various oncogenes and growth factors, partial mole appears to be more like normal placenta, while complete mole seems to be more like choriocarcinoma. These results may have both prognostic and therapeutic consequences and provide insight into the relationship between normal placenta and gestational trophoblastic diseases.