Synthesis of IFN-gamma 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-gamma(+) CD4(+), and TNF-alpha(+) CD4(+) T cells than women with HPV infection alone (HPV(+)). Whereas HIV(+) and healthy women had similar numbers of IFN-gamma(+) CD8(+) T cells, HPV(+) women had significantly fewer IFN-gamma(+) CD8(+) T cells than healthy women. CONCLUSION: HIV infection adversely affects the synthesis of Th1 cytokines by CD4(+), but not IFN-gamma synthesis by CD8(+) T cells of women with active HPV infection. The increase in IFNgamma(+) 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-gamma(+) 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.     

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     

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.     

Expression and significance of dendritic cells and Th17/Treg in serum and placental tissues of patients with intrahepatic cholestasis of pregnancy

Purpose: Dendritic cells (DCs) are involved in immune system, which can also regulate the differentiation of T helper 17 (Th17) and regulatory T cells (Treg). DCs and Th17/Treg participate in preeclampsia and recurrent spontaneous abortion (RSA), but there is still lack of research in intrahepatic cholestasis of pregnancy (ICP). The aim was to evaluate the expression and significance of CD83⁺DCs, CD1a⁺DCs, interleukin-17 (IL-17) and IL-35 in serum and placental tissues of patients with ICP.

Methods: Thirty cases of mild ICP, 25 cases of severe ICP were selected, and 30 cases of normal pregnant women were selected as control group. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) were used to detect the expression of CD83⁺DCs, CD1a⁺DCs, IL-17 and IL-35 in serum and placenta tissues, respectively.

Results: There were more CD83⁺DCs, IL-17 expressed in placenta from women with ICP than in normal pregnancies, while the number of decidual CD1a⁺DCs, IL-35 was significantly lower in ICP than in normal pregnant women. The comparison within three groups had statistical difference (p?+DCs and CD1a⁺DCs levels had no significance. IL-17 was higher in ICP, while IL-35 was lower.

Conclusions: DCs are involved in damaging the maternal–fetal immune tolerance by changing the phenotype and mature state, which may affect the differentiation of Th17/Treg to cause ICP.     

NK Cell Activity in the Presence of IL-12 Is a Prognostic Assay to Neoadjuvant Chemotherapy in Cervical Cancer

Objective. Little is known about the impact of neoadjuvant chemotherapy on cell-mediated immunity in patients with advanced cervical cancers.Patients and methods. We have studied 24 patients with advanced cervical cancer submitted to neoadjuvant chemotherapy (CT) using cis-platinum (100 mg/m²/cycle) and bleomycin (30 mg/cycle). The cell-mediated immunity parameters available before and after CT were NK cells, CD4⁺/CD28 and CD8⁺/CD28 T-lymphocyte numbers, PBMC cytotoxicity, and modification of this parameter with “in vitro” addition of IL-12.Results. The number of NK cells was higher before CT (P < 0.008) in 13 patients who presented a good clinical response to treatment, compared to 11 patients with a poor clinical response. In addition, PBMC cytotoxicity (P < 0.001), CD4⁺ and CD8⁺ T-lymphocyte values (P < 0.0047), and CD8⁺/CD28⁺ cells were also higher in the group with a good response compared with the group with a poor response. Addition of IL-12 to the medium increased the lytic capacity of PBMC after CT only in the group with a good clinical response (P < 0.05).Conclusions. NK cell numbers, CD8⁺ T-cell levels, and CD8⁺/CD28⁺ cell levels can be used as prognostic factors before CT. Our results suggest that patients with a poor response have lower lytic activity per NK cell and are refractory to IL-12 stimulation, probably as a result of the reduced expression of IL-12 receptors or of an intracellular defect in the mechanism of transduction. These observations also provide support for human clinical trials of IL-12 and neoadjuvant CT in patients with cervical cancer.     

HIV/HPV co-infection during pregnancy in southeastern Brazil: Prevalence,HPV types,cytological abnormalities and risk factors

ObjectiveHIV⁺ pregnant women are at a higher risk of HPV infection and development of cervical cancer. Our objectives were to assess the prevalence and HPV types in HIV⁺ pregnant women and to identify risk factors for HPV infection and cytological abnormalities.MethodsCervicovaginal smears were collected during pregnancy from 140 women. Partial HPV L1 gene and the exon 4 of the human TP53 gene (containing codon 72) were PCR-amplified and sequenced. Amplified products indicating multiple HPV infection were further cloned and sequenced. The association of demographic, obstetric and HIV-related clinical variables with HPV infection and cervical lesions was tested by univariate analyses, and significant factors were subsequently tested by logistic regression multivariate analysis.ResultsHPV DNA tested positive for 118 patients and HPV types were identified in 104 samples. Twenty-eight different types were found, HPV-16 and HPV-58 being the most prevalent. High-risk types were present in 79.8% of samples and multiple infections in 16.3%. Abnormal cervical smears were found in 44 patients (31.4%). Absolute CD4⁺ T-cell counts below 350 were associated with HPV infection. Younger age was associated with cervical abnormalities and higher CD4⁺ T-cell count was an apparent protective factor.ConclusionsWe found a high prevalence of HPV infection and high-risk types in this cohort. Our results highlighted the relevance of immune system integrity rather than TP53 variants for protecting this highly vulnerable population to HPV infection and carcinogenesis.     

Vulvar Intraepithelial Neoplasia in Women Infected with Human Immunodeficiency Virus-1

The purpose of this study was to determine the response of vulvar intraepithelial neoplasia (VIN) lesions to standard treatment methods in women infected with human immunodeficiency virus (HIV). We reviewed all cases of VIN over a 4-year period at an inner-city hospital. We reviewed the clinical records of these women to abstract demographic information as well as information about tobacco use, injection drug use, results of HIV testing, T cell count, stage of HIV infection, colposcopic and cytologic findings, treatment of lesions, and follow-up. Eight of the 28 women (29%) with VIN were infected with HIV. The relative risk for recurrence or persistence of VIN after treatment was 3.3 (95% confidence interval, 1.4–7.4;P= 0.01) in the HIV⁺compared with the HIV⁻group. The high rate of HIV infection among women with VIN supports recommendation of HIV testing for women with VIN. Women known to have HIV infection should be carefully examined for vulvar lesions. Further study is needed to determine the optimum treatment for VIN in women infected with HIV.     

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.     

Investigation of uterine arterial chemoembolization and uterine arterial infusion chemotherapy for advanced cervical cancer before radical radiotherapy: a long-term follow-up study

Purpose

To evaluate the immunotherapeutic potentials for human dendritic cells (DCs) loaded with different HPV16-associated antigens, including HPV16E7 (E) protein, HPV16E7 polypeptide (P), as well as CpG-oligodeoxynucleotide (ODN) 2006 as a promising immune adjuvant for vaccination against cervical carcinoma.

Methods

DCs derived from human peripheral blood and cord blood were isolated and loaded with HPV-derived protein or peptides, in combination with CpG-ODN2006 as a potential adjuvant. The IL-12 level, the allogeneic T cell-stimulatory capacity and the cytotoxicity of cytotoxic T lymphocytes (CTLs) were evaluated in vitro. Furthermore, an immune reconstitution model of human cervical carcinoma in SCID mice was used to assess the anti-tumor effects in vivo. The tumor sizes, the expression of IgG and IFN-γ, and the presence of the human CD3⁺, CD4⁺ and CD8⁺ T cells were measured in the mice inoculated with different DCs.

Results

The antigen-loaded DCs displayed obvious anti-tumor activities in vitro and in vivo, and showed no toxicity to normal cells. The level of IL-12, an important cytokine for immune response, was up-regulated in all mice inoculated with antigen-loaded DCs. Stimulation and activity of CTLs were increased after treatment with antigen-loaded DCs. Significantly, DCs loaded with HPV16E7 polypeptide (P) showed the most distinguished immunotherapeutic activities, and such effect was further enhanced when HPV16E7 polypeptide (P) was used in combination with CpG-ODN2006. Interestingly, the same results were obtained in vivo: the tumor size was decreased, and IgG and IFN-γ levels were increased after the SCID mice were inoculated with the loaded DCs.

Conclusions

HPV16E7 polypeptide combined with the immune adjuvant CpG-ODN2006 could be a suitable HPV16-associated tumor antigen. The research provides a new strategy for generating DCs vaccines for immunotherapy of cervical cancer.     

Phenotypic characterization of regulatory T cells populations in maternal blood,cord blood and placenta from diabetic mothers

Purpose: Changes in regulatory T cells (Treg) in peripheral blood are associated with a number of pathologies, including diabetes. However, the immunological responses of pregnant diabetic women remain scarcely known, and the effects of Treg cells in these patients have yet to be investigated. The present study characterized the expression of regulatory T cells in the maternal blood, cord blood and placenta of diabetic pregnant women.

Materials and methods: The women were divided according to glycemic status into a non-diabetic (ND; N?=?20) or type 2 diabetic (T2DM; N?=?20) group. Cell subsets were determined by flow cytometry.

Results: Compared to ND, T2DM blood cells exhibited a higher expression of CD25⁺, Foxp3⁺, CD4⁺CD25⁺, CD4⁺Foxp3⁺ and CD25⁺Foxp3⁺; and cord blood cells showed a lower expression of CD25⁺, CD4⁺Foxp3⁺ and CD25⁺Foxp3⁺. In the placenta of T2DM, the villous layer of the proportion, CD3⁺ and CD25 was lower than that of CD4⁺Foxp3⁺ and CD25⁺Foxp3⁺, and the extravillous placenta layer contained the lowest levels of CD4⁺ and CD25⁺ and highest proportions of CD4⁺Foxp3⁺. In maternal blood from T2DM, the frequency of CD3⁺CD95⁺ and CD3CD4⁺ T cells expressing CD95⁺ was lower. In cord blood from T2DM, the rate of CD3⁺CD95⁺ was lower. The placenta villous layer of T2DM showed a lower count of CD3⁺CD95⁺ and of CD3CD4⁺ T cells expressing CD95⁺, whereas the number of cells expressing CD3⁺CD45RO⁺ decreased in both placental layers.

Conclusion: The data obtained suggest that hyperglycemia changes the phenotypes of regulatory T cells and Fas expression in memory T cells.     

Interferon-γ and Tumor Necrosis Factor-α Induce Synergistic Cytolytic Effects in Ovarian Cancer Cell Lines—Roles of the TR60 and TR80 Tumor Necrosis Factor Receptors

Objective.Utilizing ovarian cancer cell lines, we examined the effect of IFN-γ on each type of TNF receptor. Additionally, we sought to determine the effect of receptor modulation on TNF-α-mediated cytolysis.Methods.Ovarian cancer cell lines Caov-3, A2780, and SK-OV-3 were employed. The number of TNF receptors was determined by a TNF-α binding assay utilizing¹²⁵I-labeled TNF-α. Monoclonal antibodies specific for the 55- to 60-kDa (TR60) and the 75- to 80-kDa (TR80) TNF receptors were used to determine the relative density of each receptor type. Northern blot analyses were performed employing cDNA probes for the TR60 and TR80 mRNAs. To elucidate which receptor(s) was responsible for mediating the signal for cytolysis, 24-h MTT cytolytic assays were performed in the presence of receptor-specific monoclonal antibodies.Results.IFN-γ treatment resulted in an increase in TNF receptors in the cell lines A2780 and Caov-3 (P< 0.001), but not SK-OV-3. Northern blot analyses suggested distinct regulatory mechanisms for the two receptors. In Caov-3 and SK-OV-3 cells a synergistic increase in TNF-α-mediated cytolysis was seen when cells were pretreated with IFN-γ. In both cell lines, pretreatment with IFN-γ markedly enhanced the ability of the TR60 receptor to mediate cell lysis. Conversely, under similar treatment conditions, the TR80 receptor did not appear capable of generating a cytolytic signal.Conclusions.TNF receptor modulation by IFN-γ appears to be unique to individual cell lines. The TR60 TNF receptor plays a central role in the synergistic cytolytic effects of IFN-γ and TNF-α. Sequential therapy with IFN-γ and TNF-α and specific TNF receptor activation may provide novel translational strategies for the use of cytokines in the treatment of ovarian cancer.     

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.     

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.     

Changes in T-cell phenotype and cytokines profile in maternal blood,cord blood and colostrum of diabetic mothers

Objective: The present study evaluated the cells and cytokine of maternal blood, cord blood and colostrum of diabetic mothers.

Methods: The women evaluated were divided according to their body mass index (BMI) and glycemic status into non-diabetic (ND – N?=?15), mild gestational hyperglycemic (MGH – N?=?15), diabetes mellitus gestational (DMG – N?=?13) and type-2 diabetes mellitus (DM2 – N?=?15) groups. The subsets of cells and cytokine profile were determined by flow cytometry.

Results: Maternal blood from MGH group had increase percentage of CD3⁺T cells, and DM-2 group had decrease percentage of CD4⁺ T cells. The cord blood from hyperglycemic groups showed lower percentage of CD3⁺ T cells expressing CD45RO⁺ and higher of CD4⁺ T cells and CD4⁺ T cells expressing CD45RA⁺. In the colostrum, the CD4⁺ T cells and CD4⁺ T cells expressed CD45RA⁺ increase in hyperglycemic groups. The DM2 group exhibited higher IL17 levels in maternal blood. IFN-γ was lower in cord blood from MGH and DMG groups with overweight/obese. Irrespective of the glycemic status, IL6 was higher in colostrum.

Conclusion: The results obtained suggest that maternal hyperglycemia modifies the phenotypes of T cells and cytokines profile in maternal, cord blood and colostrum.     

Changes of Th17/Tc17 and Th17/Treg cells in endometrial carcinoma

Objectives

T helper 17 (Th17), T cytotoxic 17 (Tc17) and regulatory T (Treg) cells are important factors in the pathogenesis of inflammatory and autoimmune diseases. However, information concerning the roles of these cells in antitumor immunity or endometrial tumorigenesis remains limited. In this study, we aimed to describe the distribution of Th17, Tc17 and Treg cells in endometrial carcinoma patients, and elucidate the probable role of these effector T cells.

Methods

We assessed the expression of interleukin (IL)-17 and Foxp3 in the peripheral blood of endometrial carcinoma patients and healthy controls by flow cytometry to determine the relative numbers of Th17, Tc17 and Treg cells. Th17 cells and Tc17 cells were counted as percentages of the total number of CD3⁺ T cells; Treg cells were counted as a percentage of the total number of CD4⁺ T cells. We also evaluated Th17 and Tc17 cells in tumor tissue by immunohistochemical staining. IL-17 and IL-10, dominant products of these three cell types, were detected by using enzyme-linked immunosorbent assays.

Results

The frequencies of Th17, Tc17 and Treg cells, as well as the serum level of IL-10, were significantly elevated in endometrial carcinoma patients compared to healthy controls. The Th17/Tc17 and Th17/Treg ratios were also observed to change significantly. However, there was no significant difference on the IL-17 levels in the serum. Additionally, immunohistochemistry performed on tumor tissues indicated that the amounts of Th17 and Tc17 increased in the cancer patients.

Conclusions

Our data suggests a probable involvement of Th17, Tc17 and Treg cells in the pathogenesis of endometrial carcinoma. Restoring the balance of these cells may help with the research and development of immunotherapies for endometrial carcinoma.     

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.     

Differential Response of Cervical Intraepithelial and Cervical Carcinoma Cell Lines to Transforming Growth Factor-β1

Transforming growth factor-β1 (TGF-β1) is a potent inhibitor of epithelial cell proliferation. It has been proposed that loss of sensitivity to growth inhibition by TGF-β1 may be an important step in the development of cervical carcinoma, but it remains unclear whether this represents an early or a late event. We compared the sensitivity to TGF-β1 of nontumorigenic human papillomavirus deoxyribonucleic acid (HPV DNA)-positive cell lines derived from cervical intraepithelial neoplasia (CIN), of newly established cervical carcinoma cell lines, of nontumorigenic HPV DNA-transfected cervical cell lines, and of normal ectocervical cells. There is a dose-dependent inhibition of DNA synthesis by TGF-β1 in the CIN cell lines and the HPV DNA-transfected cell lines. The carcinoma cell lines are resistant to the growth inhibitory effects of TGF-β1. The CIN cell lines are significantly more sensitive than the carcinoma cell lines (P < 0.001), but significantly less sensitive than normal cervical cells (P < 0.05). A CIN cell line which contains HPV 31b DNA is more sensitive to TGF-β1 at early passage than at late passage (P < 0.05). There are no differences in the sensitivity to the growth inhibitory effects of TGF-β1 between subclones of this cell line that have different episomal HPV DNA content, population-doubling time, or differentiation characteristics. Both normal and abnormal cervical epithelial cells were able to secrete latent TGF-β1 or TGF-β2. We conclude that resistance to growth inhibition by TGF-β1 is likely to be a late event in the development of cervical carcinoma; it is not the mere consequence of immortalization by HPV genes acquired following transfection in vitro or infection in vivo.     

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.     

Hypertension,inflammation and T lymphocytes are increased in a rat model of HELLP syndrome

Objective: An animal model of hemolysis, elevated liver enzymes, low platelet count (HELLP) was used to determine if T lymphocytes accompany hypertension and increased inflammatory cytokines. Methods: sFlt-1 (4.7?µg/kg/day) and sEndoglin (7?µg/kg/day) were infused into normal pregnant rats (HELLP rats) for 8 days. Results: HELLP was associated with increased mean arterial pressure (p?=?0.0001), hemolysis (p?=?0.044), elevated liver enzymes (p?=?0.027), and reduced platelets (p?=?0.035). HELLP rats had increased plasma levels of TNFα (p?=?0.039), IL-6 (p?=?0.038) and IL-17 (p?=?0.04). CD4⁺ and CD8⁺ T lymphocytes were increased. Conclusion: These data support the hypothesis that T cells are associated with hypertension and inflammation.