Synthesis of IFN-γ by CD8 T Cells Is Preserved in HIV-Infected Women with HPV-Related Cervical Squamous Intraepithelial Lesions

Objective. The aim of this study was to investigate whether coinfection with HIV affects the synthesis of Th1 and Th2 cytokines by peripheral blood T cells of women infected with human papillomavirus (HPV).Methods. Cervical swabs and peripheral blood were obtained from women referred for colposcopy. HPV DNA by Digene's hybrid capture assay, HIV RNA by Roche's Amplicor assay, and cytokine synthesis of T-cell subsets by flow cytometry were assessed. HPV-associated cervical and HIV-associated immune deficiency diseases were staged using the Bethesda System and the Centers for Disease Control criteria, respectively.Results. Patients with HIV and/or HPV infections had lower percentages of IL-2⁺ and higher percentages of IL-10⁺ T cells than healthy women. Furthermore, women with both virus infections (HIV⁺/HPV⁺) had significantly fewer IL-2⁺ CD4⁺, IFN-γ⁺ CD4⁺, and TNF-α⁺ CD4⁺ T cells than women with HPV infection alone (HPV⁺). Whereas HIV⁺ and healthy women had similar numbers of IFN-γ⁺ CD8⁺ T cells, HPV⁺ women had significantly fewer IFN-γ⁺ CD8⁺ T cells than healthy women.Conclusion. HIV infection adversely affects the synthesis of Th1 cytokines by CD4⁺, but not IFN-γ synthesis by CD8⁺ T cells of women with active HPV infection. The increase in IFNγ⁺ CD8⁺ T cells, a phenotype consistent with cytotoxic T lymphocytes, may account for the stable HIV disease of the women studied. However, the increase in IFN-γ⁺ CD8⁺ T cells is less likely to be HPV-specific as there was a higher incidence of HPV-related cervical SIL in HIV⁺/HPV⁺ women compared with HPV⁺ women.     

Intraperitoneal immune cell status in infertile women with and without endometriosis

Endometriosis is a widespread chronic disease characterized by endometrial tissue located outside the uterine cavity. Clinical signs are chronic pelvic pain and infertility. Emerging evidence indicates that the immune system is profoundly involved in the onset and/or progression of endometriosis. However, mechanistic pathways have not yet been conclusively specified. In this study, women undergoing diagnostic laparoscopy due to infertility were recruited, and classified as early-stage endometriosis (n = 30), advanced-stage endometriosis (n = 8) or no endometriosis (n = 31). The frequency and phenotype of leukocytes were evaluated in peritoneal fluid. While the frequency of lymphocytes was not significantly different, neutrophils were increased in endometriosis. Flow cytometry analysis revealed an increased frequency of CD4⁺ and CD8⁺ cells in peritoneal fluid of endometriosis patients. In addition, the frequency of CD4⁺CD25⁺CD103⁺ cells and lineage⁻HLA-DR⁺CD11c⁺CD123⁺ dendritic cells was decreased in peritoneal fluid in endometriosis, whereas CD57⁺ NK cells and CD8⁺CD28⁻ T suppressor cells remained largely unaltered. We conclude that therapeutic approaches in endometriosis might focus on peritoneal leukocytes as a target or surveillance marker; however, immune alterations in peritoneal fluid are subtle and their analysis will require highly standardized and harmonized protocols.     

High Serum Levels of Soluble IL-2 Receptor, Cytokines, and C Reactive Protein Correlate with Impairment of T Cell Response in Patients with Advanced Epithelial Ovarian Cancer

The serum levels of interleukin-(IL-)1α, IL-1β, IL-2, IL-6, TNFα, and sIL-2R and the proliferative response of peripheral blood mononuclear cells (PBMC) to phytohemagglutinin (PHA), anti-CD3 monoclonal antibody (mAb), recombinant IL-2 (rIL-2), and the combination of PHA or anti-CD3 mAb with rIL-2 were studied and correlated with serum levels of C-reactive protein (CRP) in women with advanced epithelial ovarian cancer. The expression of CD25 and CD122 subunities of membrane-bound IL-2R on PHA- or anti-CD3 mAb-stimulated PBMC was also studied. In comparisons with the controls, PBMC response to PHA, anti-CD3 mAb, and rIL-2 was significantly lower in the cancer patients. The addition of exogenous rIL-2 to the PBMC cultures increased response in both controls and patients but did not modify the significance of the differences. After stimulation with PHA or anti-CD3 mAb, the percentage of PBMC CD25⁺or CD122⁺was significantly lower in patients. The serum levels of IL-1α, IL-1β, IL-6, TNFα, sIL-2R, and CRP were significantly increased in patients compared to the controls. Instead, no differences were observed for serum levels of IL-2. A strong association was found between high serum levels of the above-mentioned cytokines, sIL-2R, and CRP. The results of our study on advanced stage (IIIb–IV) ovarian cancer patients are consistent with the previously reported hypothesis that high IL-6 and/or CRP serum levels may represent an important and independent prognostic factor of the likely outcome in cancer patients.     

Decreased frequency of peripheral blood CD8+CD25+FoxP3+regulatory T cells correlates with IL-33 levels in pre-eclampsia

Objective: We aimed to investigate the role of CD8⁺CD25⁺Foxp3⁺regulatory T (Treg) cells in pre-eclampsia (PE).

Methods: This was a cross-sectional study of 46 patients with PE and 24 normotensive women within the third trimester of gestation. We analyzed the percentages of CD8⁺CD25⁺Foxp3⁺Treg cells in peripheral blood using flow cytometry and the serum levels of interleukin (IL)-6, IL-17A, IL-10, TGF-β1, IL-1β, and IL-33 by Luminex 200.

Results: We found that patients with PE had lower percentages of CD8⁺CD25⁺Foxp3⁺Treg cells than normotensive pregnant women. In addition, the percentage of CD8⁺CD25⁺Foxp3⁺Treg cells was positively correlated with IL-33 concentration and negatively correlated with IL-17A concentration in patients with PE. We also found that IL-33 treatment can induce proliferation of CD8⁺CD25⁺Foxp3⁺Treg cells in vitro.

Conclusions: These findings suggest that the reduced CD8⁺CD25⁺Foxp3⁺Treg cells may play a role in the pathogenesis of PE.

Abbreviations

PE: pre-eclampsia; PBMCs: peripheral blood mononuclear cells; CTLA-4: cytotoxic T-lymphocyteantigen-4; APCs: antigen presenting cells; TGF-β: transforming growth factor-β; IL: interleukin; Treg: cells regulatory T cells; PBS: phosphate-buffered saline; Foxp3: forkhead Box protein 3; HELLPs: hemolysis, elevated liver enzyme and low platelet syndrome     

Altered distribution of NK and NKT cells in follicular fluid is associated with IVF outcome

The aim of this study was to investigate differences in the relative distributions of subsets of natural killer (NK) cells, including immunoregulatory NK cells (CD56⁺CD16⁻), cytotoxic NK cells (CD56⁺CD16⁺), as well as total NK cells (CD56⁺CD3⁻), and NKT cells (CD56⁺CD3⁺) in peripheral blood and follicular fluid in subjects with successful or unsuccessful IVF treatment. The immunoregulatory NK cell population in follicular fluid of women who failed to achieve pregnancy after IVF treatment was significantly decreased compared to women who became pregnant after IVF. Conversely, the NKT cell population in the follicular fluid of women with unsuccessful treatment was significantly elevated compared with those with successful IVF. Understanding the changes in the distribution of NK and NKT cell populations in follicular fluid might serve as the basis for a more detailed study to determine whether NK cell parameters have prognostic value in guiding the selection of individual ova for use in IVF procedures.     

Erythropoietin Treatment under Polychemotherapy in Patients with Gynecologic Malignancies: A Prospective, Randomized, Double-Blind Placebo-Controlled Multicenter Study

In order to examine the influence of erythropoietin (rHuEPO) on serum hemoglobin levels, transfusion requirements, and quality of life in patients with gynecologic malignancies under polychemotherapy and chronic tumor anemia (hemoglobin <11 g/dl), we performed a prospective, randomized, double-blinded placebo-controlled clinical trial. Between October 1992 and October 1993, 35 patients from 5 gynecologic departments were entered into this trial. Inclusion criteria were hemoglobin level <11 g/dl, ferritin level >29 ng/ml, stool negative for occult blood, and life expectancy for more than 3 months. Patients received either 150 U/kg body wt rHuEPO (Erypo by Cilag-Janssen) sc three times a week for 12 weeks (n= 23) or a placebo (n= 12). If the hemoglobin levels of the 4th, 8th, or 12th week were >2 g/dl above the baseline value and/or >12 g/dl, the patient was classified as a responder. Patients who required blood transfusions (hemoglobin <8 g/dl, erythrocytes <3 × 10⁶/ml, or clinical symptoms of anemia) were classified as nonresponders. A nonvalidated quality of life questionnaire was completed by the patient at the beginning of the treatment and then every fourth week before receiving chemotherapy. In the rHuEPO group 56.6% of the patients responded to the treatment (χ²= 10.79,P= 0.001) and only 5 patients (21.7%) required blood transfusions, whereas 8 of 12 patients in the placebo group (66.6%) had to be transfused (χ²= 6.81,P= 0.009). Quality of life did not differ significantly between the rHuEPO group and the placebo group of patients. Within the rHuEPO group those patients that responded showed a significant increase in physical activity after response in comparison to the preresponsive phase (P= 0.02, pairedttest). We therefore concluded that rHuEPO significantly increases serum hemoglobin levels and decreases transfusions requirements while maintaining quality of life in patients with gynecological malignancies who are undergoing polychemotherapy.     

Histology/Cytology Discrepancies in HIV-Infected Obstetric Patients with Normal Pap Smears

Objective.To estimate the frequency of cervical cytologic/histologic discrepancies in a group of obstetric patients diagnosed as HIV infected by routine prenatal screening. Also, to determine if serum CD4 levels or sexually transmitted diseases (STDs) are associated with the occurrence of preinvasive cervical disease in these women.Method.Thirty-two women who presented for routine prenatal care to Medical Center of Louisiana were diagnosed as HIV infected by ELISA and Western blot testing and had normal Pap smears. These patients then agreed to undergo the following: colposcopy with directed biopsies; chlamydia, gonorrhea, and syphilis screening; and serum CD4 level.Results.No patients had AIDS-defining diagnoses other than CD4 < 200/mm³. Ten of 32 (31%) had cervical intraepithelial neoplasia (CIN) despite normal cytology. Six of 32 (19%) had STDs. One of 10 in the group with CIN had a STD. The mean CD4 level in those patients with CIN was 249/mm²(range 1–524) vs 501/mm²(range 210–979) in those without CIN. (P= 0.0118)Conclusions.Newly diagnosed HIV-infected pregnant women without clinical evidence of AIDS are noted to have CIN at a rate similar to nonpregnant HIV-infected women. The Pap smear appears to have a significant false-negative rate in this group. STDs, while common, were not directly associated with false-negative Pap smears. CIN is associated with immunosuppression, as measured by low CD4 counts.     

Decidual CD4+CD25+CD127dim/- regulatory T cells in patients with unexplained recurrent spontaneous miscarriage

Objectives

To investigate the frequency and function of CD4⁺CD25⁺CD127^dim/− regulatory T (Treg) cells in decidua of patients with unexplained recurrent spontaneous miscarriage (URSM).

Study design

The decidual lymphocytes from patients who experienced URSM and normal pregnant women (controls) were collected by Ficoll density gradient centrifugation. CD4⁺CD25⁺CD127^dim/− Treg cells were isolated by magnetic cell sorting. The proportion of Treg cells and IL-10, TGF-β in Treg cells were determined by flow cytometry. Inhibitory effects of Treg cells on effecter T cells were detected with or without the presentation of anti-IL-10 antibodies and anti-TGF-β antibodies.

Results

The frequency of CD4⁺CD25⁺CD127^dim/− Treg cells was decreased in URSM decidua compared to controls (2.09% ± 0.86% vs. 2.97% ± 1.19%, p = 0.005), and the expression of IL-10 and TGF-β in Treg cells was lower in the URSM group than in the control group (p = 0.04 and p = 0.01, respectively). Furthermore, the suppressive effect of CD4⁺CD25⁺CD127^dim/− Treg cells on the proliferation of effector T cells was decreased in URSM decidua (p < 0.05). Suppression was mediated predominantly through IL-10 and TGF-β in decidua.

Conclusions

Decreased frequency and immunosuppressive capacity of CD4⁺CD25⁺CD127^dim/− Treg cells was found in URSM decidua. Treg cells inhibit proliferation of effector T cells mainly via IL-10 and TGF-β in URSM decidua.     

Viability and functional capacity after thawing of hematopoietic progenitor cells cryopreserved at a cord blood stem cell bank in Colombia

Objective

To evaluate the viability and functional capacity of hematopoietic progenitor cells from cord blood samples cryopreserved at the Banco de Células Stem de Colombia.

Methods

After thawing and centrifugation of 20 samples, viable white blood cells were numbered by the trypan blue method and CD34⁺CD45^+ dim hematopoietic progenitor cells were numbered by flow cytometry. Clonogenic assays also tested the functional capacity of viable CD34⁺CD45^+ dim cells.

Results

The median rates of viable CD34⁺CD45^+ dim cells were 99.6% before freezing and 73.0% after thawing (P < 0.001). The 20 cultures yielded a median of 12 cells with a lineage of red cells, 17.5 cells with a lineage of white cells, and 10 cells with a mixed lineage.

Conclusion

Although the rate of viable CD34⁺CD45^+ dim cells was decreased by 26.6% after thawing by the method we used, the numbers of CD34⁺CD45^+ dim cells that formed colonies were similar to those obtained by other published methods.     

Effect of TLR3 and TLR7 activation in uterine NK cells from non-obese diabetic (NOD) mice

Toll-like receptor (TLR)–TLR cross talk is thought to be important in TLR signaling. Herein, we investigated the effect of specific TLR3 and TLR7 agonists, poly (I:C) and R837, individually and in combination, on uterine immune cell function and their subsequent effects on pregnancy outcome. Allogeneic pregnancies in the non-obese diabetic (NOD) mouse × C57BL/6 and wild-type BALB/c × C57BL/6 model were used. An additive increase in embryo resorption was observed after induction with both poly (I:C) and R837, and was associated with elevated numbers of both TNF-α- and IFN-γ-producing CD45⁺ cells in the uterus. Further examination showed that while cytokine expression was detected in both CD3⁺ cells and CD49b⁺ cells in BALB/c mice, NOD mouse cells behaved differently. In NOD mice, elevated cytokine expression was attributed to CD3⁺ T cells, with no response detected in the CD49b⁺ NK cells. The additive effect of combined agonists was partially inhibited by the Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) inhibitor SP600125 and almost completely abrogated by the extracellular signal-regulated kinase (ERK) MAPK inhibitor PD98059. These results suggest that increased TLR3 and TLR7 signals are transmitted via Th1-type T cells, rather than NK cells, in NOD mice. Furthermore, the ERK MAPK pathway may be critical in TLR3 and TLR7 signaling.     

Cold-Knife Conization versus Loop Excision: Histopathologic and Clinical Results of a Randomized Trial

Ninety patients with cervical intraepithelial neoplasia (CIN) were randomly assigned to loop excision (n = 38) or cold-knife conization (n = 52). All specimens were well evaluable at histology. The average width of the lesions at histology was 10.2 and 9.7 mm, respectively (ns). The average weight of the specimens was 2.6 and 5.6 g (P < 0.01) and the average depth was 9.2 and 15.8 mm (P < 0.01), respectively. The distance between the cervical resection margin and CIN was 14 mm after loop excision and 24 mm after cold-knife conizatiun (P < 0.06). The margins of the specimen were not clear of disease in 8 patients after loop excision and in 12 patients after conization (ns). Two patients after loop excision and in three patients after cold-knife conization had postoperative bleeding. The results suggest that, compared with cold-knife conization, loop excision removes less healthy tissue without reducing the chances for cure.     

Characterization of decidual leukocyte populations in cynomolgus and vervet monkeys

The objective of this study was the phenotypic and functional evaluation of decidual immune cells in the cynomolgus and vervet monkeys. Early pregnancy (days 36–42) deciduas were obtained by fetectomy for histological evaluation and decidual mononuclear leukocyte (MNL) isolation. While peripheral NK (pNK) cells in these species do not express CD56, CD56⁺ NK cells were abundant in decidual samples. The majority of decidual NK (dNK) cells (>80%) had high light-scatter characteristics and were CD56^brightCD16⁺ cells with no or very low levels of natural cytotoxicity receptors (NKp46, NKp30) and NKG2A, while a minor population were small CD56^dimCD16⁻ lymphocytes also expressing less NKp46, NKp30 and NKG2A than pNK cells. All dNK cells were found to be perforin⁺; however, their cytotoxic potential was low and cynomolgus dNK cells showed strongly reduced cytotoxicity against target cells compared with pNK cells. Macrophages and T cells together comprised approximately 25–30% of decidual MNL. Decidual T cells contained a higher proportion of the minor T cell subtypes (γδT cells, CD56⁺ T cells) compared with peripheral blood. A subset of DC-SIGN⁺ macrophages, with a distribution adjacent to areas of placental attachment in contrast to the widespread setting of general CD68⁺ cells, was identified in both species. Together, these results demonstrate that the maternal–fetal interface in both cynomolgus and vervet monkeys is very rich in immune cells that have similar phenotypes to those seen in humans, indicating that both species are excellent models to study the contributions of distinct immune cell populations to pregnancy support.     

High-Dose Platinum versus Standard Dose in Advanced Ovarian Carcinoma: A Randomized Trial from the Gynecologic Cooperative Group of the French Comprehensive Cancer Centers (FNCLCC)

Objective. The aim of this study was to evaluate the impact of platinum dose intensity on pathological response rate and overall survival in patients with advanced ovarian adenocarcinoma.Methods. Between February 1992 and December 1996, 195 previously untreated patients with FIGO stage IIb-c, IIIb-c, or IV with macroscopic residual disease after suboptimal debulking surgery were randomized to receive CCC (100 mg/m² of cisplatin, 300 mg/m² of cyclophosphamide, 300 mg/m² of carboplatin, n = 96) or CC (100 mg/m² of cisplatin, 600 mg/m² of cyclophosphamide, n = 99) for six courses at 28-day intervals. A second-look laparotomy was planned at the end of chemotherapy.Results. In the CCC arm, the platinum compound received dose intensity and relative total dose were 85 and 76%; in the CC arm, they were 94 and 85%. Grade 3–4 toxicity was more frequent in the CCC arm than in the CC arm for leukopenia (56% vs 26%, P < 0.001), febrile neutropenia (18% vs 4%, P = 0.002), anemia (31% vs 5%, P < 0.001), thrombopenia (55% vs 4%, P < 0.001), and ototoxicity (8% vs 0%, P < 0.001). The pathologic complete response rate was 22 and 14% in the CCC and CC arms, respectively (P = 0.19). With a median follow-up of 53 months, the median time to failure and the 3-year treatment failure-free survival rate were 17.4 months and 22% vs 13 months and 11% in the two arms, respectively (P = 0.01). The median survival time and the 3-year overall survival rate were, respectively, 30 months and 42% vs 25 months and 33% (P < 0.20).Conclusion. The platinum dose intensification (1.6-fold increase) obtained with the CCC association improves the treatment failure-free survival without significant impact on overall survival when compared with the CC regimen in suboptimal debulked ovarian adenocarcinoma. However, because of its high rate of hematologic toxicity and ototoxicity, this association cannot be recommended for routine practice.     

Preoperative Ultrasound Study in Predicting Lymph Node Metastasis for Endometrial Cancer Patients

Objective.The objective of this study was to evaluate the efficacy of preoperative ultrasound (US) findings such as tumor size, status of myometrial invasion, and intratumoral “resistance index” (RI) in predicting lymph node metastasis in endometrial carcinoma patients.Methods.Forty-two patients with endometrial cancer were enrolled. All patients underwent total abdominal hysterectomy, pelvic lymph node dissection or sampling, and para-aortic lymph node sampling. Two-dimensional and color Doppler US were performed before surgery to measure tumor size, depth of myometrial invasion, and intratumoral arterial RI. Formalin-fixed, paraffin-embedded pathologic slides from surgical specimens were reviewed by a senior pathologist to evaluate histologic type and grade, depth of myometrial invasion, cervical involvement, lymph-vascular emboli, and status of lymph node metastasis.Results.There were 12 patients with pelvic and/or para-aortic lymph node metastases and 30 patients without nodal metastases. Patients with tumors larger than 2.5 cm by US (11/12 vs 14/30,P= 0.008), more than half myometrial invasion by US (9/12 vs 5/30,P< 0.001), and intratumoral RI values less than 0.4 by US (12/12 vs 4/30,P< 0.001) had a significantly higher incidence of nodal metastases as compared with patients with tumors smaller than 2.5 cm, no or superficial myometrial invasion, and RI values higher than 0.4, respectively. Multiple regression analysis showed that only intratumoral RI values less than 0.4 were significantly correlated with nodal metastasis (P< 0.001,r²= 0.650). We used the intratumoral RI value as the parameter to evaluate nodal metastasis in endometrial cancer patients. Twelve of sixteen patients with intratumoral RI values <0.4 had a high incidence of nodal metastases. None of the 26 patients with intratumoral RI values >0.4 had nodal metastases.Conclusions.Preoperative ultrasound features can offer important information for predicting lymph node metastasis in endometrial cancer patients. Patients with tumors with intratumoral RI values less than 0.4 should be highly suspected of having lymph node metastases and further management such as pelvic lymph node dissection or postoperative pelvic radiotherapy would be needed for these patients.     

Longitudinal study of CD4+ cell counts in HIV-negative pregnant patients

Objective.?To evaluate the absolute CD4⁺, CD8⁺, and lymphocyte cell counts and percentages from the first trimester through 6–12 weeks post-delivery in normal human immunodeficiency virus (HIV)-negative pregnant patients.

Methods.?A longitudinal laboratory analysis was performed during pregnancy that involved 51 HIV-negative subjects with blood analysis obtained in all trimesters, at delivery, and 6–12 weeks post-delivery. Twenty-five HIV-negative non-pregnant controls were also evaluated. Blood was analysed for absolute CD4⁺, CD8⁺, and lymphocyte cell counts and percentages. Means, standard deviations, trends, and differences were examined.

Results.?The mean white blood cell (WBC) count is elevated above the non-pregnant state and this parameter increases through the pregnancy up to and including parturition. The mean absolute lymphocyte cell count, lymphocyte percentage, and absolute CD4⁺ cell count are significantly lower during pregnancy and the progression through pregnancy appears U-shaped. The mean absolute CD8⁺ cell count is not significantly different. The CD4⁺ and CD8⁺ percentages are higher during pregnancy and this elevation persists into the 6–12 week post-delivery time period. A 3-digit drop in CD4⁺ percentage is common during pregnancy between blood draws; whereas, a 30% decrease or more in absolute CD4⁺ cell count is rare.

Conclusions.?By longitudinal analysis, pregnancy appears to significantly elevate the mean values of the WBC count, CD4⁺ percentage, and CD8⁺ percentage, but significantly decreases the absolute lymphocyte count, lymphocyte percentage, and absolute CD4⁺ cell count when compared to non-pregnant controls. The mean absolute CD8⁺ cell count appears to be unaffected.     

Prognostic Significance of Epithelial–Stromal Vascular Cuffing and Microvessel Density in Squamous Cell Carcinoma of the Uterine Cervix

Objective. Tumor angiogenesis has been shown to play an important role in tumor growth and metastasis. This study examines the prognostic significance of two histological markers of angiogenesis, i.e., vascular cuffing (VC), a bead-like arrangement of microvessels closely surrounding microscopic tumor nests, and microvessel density (MVD), the number of microvessels in a unit area, in cervical squamous cell carcinoma.Methods. One hundred twenty-two specimens from surgically resected uteri with cervical squamous cell carcinoma were histologically reviewed and immunostained for CD34. VC was graded into “none,” “incomplete,” and “complete.” The MVD was determined by counting the microvessels with a light microscope within a ×200 field area where neovascularization occurred most actively. Stromal inflammation was also split into three grades. The relationship of VC or MVD to clinicopathological prognostic factors such as FIGO stage, cervical stromal invasion, lymph–vascular space invasion, pelvic lymph node metastasis, and parametrial invasion was evaluated using univariate and multivariate analyses.Results. The patients with a complete VC pattern showed a significantly worse prognosis compared to those with a pattern graded as either none or incomplete (P < 0.011 and P < 0.0001, respectively). The Cox regression analysis revealed the complete VC pattern, together with parametrial invasion, to be an independent prognostic indicator for overall survival. MVD and the grading of stromal inflammation showed no significant relationship with VC or overall survival.Conclusions. The complete VC pattern may therefore be a useful prognostic indicator in cervical squamous cell carcinoma.     

Maternal PD-1 regulates accumulation of fetal antigen-specific CD8 T cells in pregnancy

The failure to reject the semi-allogeneic fetus suggests that maternal T lymphocytes are regulated by potent mechanisms in pregnancy. The T cell immunoinhibitory receptor, Programmed Death-1 (PD-1), and its ligand, B7-H1, maintain peripheral tolerance by inhibiting activation of self-reactive lymphocytes. Here, we investigated the role of the PD-1/B7-H1 pathway in maternal tolerance of the fetus. Antigen-specific maternal T cells both proliferate and upregulate PD-1 in vivo at mid-gestation in response to paternally inherited fetal antigen. In addition, when these cells carry a null deletion of PD-1, they accumulate excessively in the uterus-draining lymph nodes (P < 0.001) without a concomitant increase in proliferation. In vitro assays showed that apoptosis of antigen-specific CD8⁺ PD-1^−/− cells was reduced following peptide stimulation, suggesting that the accumulation of these cells in maternal lymph nodes is due to decreased cell death. However, the absence of neither maternal PD-1 nor B7-H1 had detectable effects on gestation length, litter size, or pup weight at birth in either syngeneic or allogeneic pregnancies. These results suggest that PD-1 plays a previously unrecognized role in maternal–fetal tolerance by inducing apoptosis of paternal antigen-specific T cells during pregnancy, thereby controlling their abundance.     

Immunohistochemical expression of Annexin A2 and S100A proteins in patients with bulky stage IB-IIA cervical cancer treated with neoadjuvant chemotherapy

Objective

Abnormal expression of Annexin A2 and S100A proteins has been reported to induce sensitivity/resistance to chemotherapy in a variety of cancers. The aim of this study was to evaluate the significance of Annexin A2 and S100A protein expression to predict response to neoadjuvant chemotherapy and prognostic significance of these protein expressions in bulky stage IB-IIA cervical cancer patients.

Methods

Paired tumor samples (pre- and post-chemotherapy) were obtained from 68 patients who were treated with cisplatin-based neoadjuvant chemotherapy and radical hysterectomy at our hospital from 2006 to 2011. The expression of Annexin A2 and S100A proteins was analyzed by immunohistochemistry.

Results

Thirty-six patients were identified as chemotherapy-response and 32 were non-response. (a). Protein expression in tumor cells: (1). Exposure of tumor cells to chemotherapy results in a change of Annexin A2 and S100A expression (P < 0.05). (2). Annexin A2, S100A8 and S100A9 protein expression correlates with tumor response to chemotherapy (P < 0.05). (b). Protein expression in stromal cells: (1). Expression of Annexin A2, S100A8 and S100A9 was increased, but S100A2 and S100A4 was decreased after exposure to chemotherapy (P < 0.05). (2). Only S100A4 expression was associated with response to chemotherapy (P < 0.05). Multivariate analysis revealed that tumor size (P = 0.022), differentiation (P = 0.000), Annexin A2 expression in stromal cells (P = 0.009), and S100A8 expression in tumor cells (P = 0.008) were independent prognostic factors for progression-free survival of cervical cancer patients.

Conclusions

Expression of some of the measured proteins in tumor and stromal cells correlates with chemotherapy exposure, response to therapy, and progression-free survival.     

Tumor Response to Neoadjuvant Chemotherapy Correlates with the Expression of P-Glycoprotein and PCNA but Not GST-π in the Tumor Cells of Cervical Carcinoma

Objective.To identify the clinicopathological and chemoresistant factors predicting the response to neoadjuvant chemotherapy and the patient prognosis in high-risk cervical carcinomas.Methods.We retrospectively reviewed 47 patients with locally advanced or bulky cervical carcinoma treated with two courses of intraarterial infusion of cisplatin, doxorubicin, mitomycin C, and 5-fluorouracil (5-FU), followed by radical hysterectomy at our hospital between 1988 and 1995. Expressions of the chemoresistance-related proteins, such as P-glycoprotein, glutathioneS-transferase π (GST-π), and proliferating cell nuclear antigen (PCNA) in the tumor cells, were examined by immunohistochemistry using pretreatment biopsy specimens. These results were compared with the chemotherapeutic response, which was evaluated by magnetic resonance imaging (MRI) and histopathology. Outcome of the patients was also studied.Results.Chemotherapeutic effect of either complete (CR) or partial (PR) response on MRI was obtained in 36 of the 47 (86%) patients. Poor response to chemotherapy was significantly correlated with P-glycoprotein expression (P< 0.005) and low PCNA labeling (P< 0.05), but not GST-π expression in the tumor cells. Independent prognostic factors for patient survival were parametrial involvement and lymph node metastasis. Neither the expression of GST-π nor PCNA was correlated with the patient survival.Conclusion.Assessment of the expression of P-glycoprotein and PCNA is potentially useful for the prediction of tumor response to neoadjuvant chemotherapy for cervical carcinomas.