Changes in the immunophenotype of endometrium during implantation window receptivity formation in healthy fertile women.

Introduction: Endometrial receptivity is largely determined by the immunophenotype of endometrium, especially uterine NK-cells (uNK). Immune component is directly involved in the formation of favourable microenvironment for the blastocyst implantation and placenta formation, but the way it changes during the maturation of endometrial tissue in healthy fertile women is still underexplored.

Methods: The endometrium was collected from 47 healthy oocyte donors after controlled ovarian stimulation: 23 women on the day of oocytes retrieval (OR) and 24 women on the term of implantation window (IW).

A Blinded Investigation: Accentuated NK Lymphocyte CD335 (NKp46) Expression Predicts Pregnancy Failures.

Aim: NKp46 is an NK cell receptor uniquely expressed by NK cells and a small subset of innate lymphoid cells. In our previous studies, we suggested a tight connection between the activity of NK cells and the expression of NKp46 and supported the clinical significance of NKp46 expression in NK cells in women with reproductive failures. In this study, we investigated the expression of NKp46 in NK cells in the peripheral blood of women in early pregnancy and analyzed its association with pregnancy loss.

A blinded multicenter investigation: Accentuated NK lymphocyte CD335 (NKp46) expression predicts reproductive failures after IVF.

The peripheral blood NK cell diversity is highly complex. Recent studies have described more than a thousand phenotypes sharing NK cell receptors (NKRs), across the leukocyte lineages. Previously, we have found that accentuated NK p46 phenotype has prognostic value for NK cytotoxicity status, and is characteristic for patients with recurrent implantation failure (RIF).

Accentuated Peripheral Blood NK Cytotoxicity Forms an Unfavorable Background for Embryo Implantation and Gestation.

Problem Many studies have demonstrated the negative impact of high rates of NK cytotoxicity (NKc) on reproductive processes, but there is no agreement as to the appropriateness and feasibility of using the NKc for routine diagnostic in IVF patients. This study conducted a retrospective analysis of embryo transfer (ET) success rates and live birth rates (LBR) in patients with different NKc values. Method of study 1854 cycles of ET were selected and divided into three groups according to NKc levels, and randomized by anamnesis and age: normal (nNKc, n = 871), elevated (eNKc, n = 759), and reduced NKc (rNKc, n = 123).

Natural killer frequency determines natural killer cytotoxicity directly in accentuated zones and indirectly in "moderate-to-normal frequency" segment.

Natural killer (NK) frequency and NK cytotoxicity (NKc) are key determining factors of a clinical outcome. In our previous study, we showed the prognostic clinical significance of immune parameters when they are beyond the optimal range (accentuated). In this study, we attempted to explain the disparity of accentuated but physiologically and immunologically normal NK parameters that might serve as negative clinical prognostics indications of failed pregnancies.

Expression of natural cytotoxicity receptor NKp46 on peripheral blood natural killer cells in women with a history of recurrent implantation failures.

Aim: The peripheral blood natural killer (NK) cells diversity is highly complex; recent studies described more than a thousand phenotypes sharing NK cell receptors, across the leukocyte lineages. In this study, we investigated the expression of NKp46 in peripheral blood NK cells in women with a history of recurrent implantation failures (RIF) with euploid embryos with pre-implantation genetic diagnosis (PGD) and control group (donors of oocytes and surrogate mothers).

Methods: The expression of NKp46 in peripheral blood lymphocytes and NK cells from women with RIF (n = 57) and control group (n = 50) was analyzed with three-color flow cytometry.

Beyond Uterine Natural Killer Cell Numbers in Unexplained Recurrent Pregnancy Loss: Combined Analysis of CD45, CD56, CD16, CD57, and CD138.

Changes in the number and cytotoxic potential of uterine Natural Killer (uNK) cells have been associated with reduced fertility. To provide a better characterization of immunophenotypes in the endometrium of women with uRPL (unexplained recurrent pregnancy loss), we examined the applicability of a set of five immune cell markers. The concentration (cells/mm) of CD45 leukocytes, CD56 uNK cells, and CD138 plasma cells as well as of CD16 and CD57 cells, which indicate high cytotoxic uNK cells, were assessed by immunohistochemistry in endometrial biopsies from 61 uRPL patients and 10 controls.

Comparison of T and NK lymphocyte subsets between human endometrial tissue and peripheral blood.

Immune profiles in endometrium may be changed in patients with IVF failure and its possible correlations with immune parameters in peripheral blood are important for the diagnostic approach. Such correlations in healthy women are unknown and have been studied in the present research. The expression of CD56, CD158a, HLA DR, CD69 in T lymphocytes, CD4 T lymphocytes, CD8+ T lymphocytes and NK cells were studied by flow-cytometry in endometrium and peripheral blood in healthy 24 donors of oocytes aged 25-32 years.

HCMV seropositivity is associated with specific proinflammatory immune phenotype in women with implantation failure.

HCMV coevolved with humans for millions of years and is now one of the most widespread infections worldwide. For a long time HCMV seropositivity was considered a safe clinical condition. In recent decades both clinical observations and research results indicated that the very presence of HCMV in human organism specifically influences immune system and may affect reproduction as a process greatly dependent on immune cells function.

Enumeration of peripheral blood NKp46 positive NK lymphocytes reflects NK cytotoxic activity in vitro.

Natural killer (NK) cells are the predominant innate lymphocyte subsets that mediate anti-tumor and anti-viral responses. The monitoring of NK cells function is important in various physiological and pathological conditions. Different approaches have been used to directly or indirectly evaluate NK cells activities.

shRNA-Induced Knockdown of a Bioinformatically Predicted Target IL10 Influences Functional Parameters in Spontaneously Hypertensive Rats with Asthma.

One of the most common comorbid pathology is asthma and arterial hypertension. For experimental modeling of comorbidity we have used spontaneously hypertensive rats with ovalbumin (OVA)-induced asthma. Rats were randomly divided into three groups: control group, OVA-induced asthma group; OVA-induced asthma + IL10 shRNA interference group.

Surface Plasmon Resonance Investigations of Bioselective Element Based on the Recombinant Protein A for Immunoglobulin Detection.

The developed surface plasmon resonance (SPR) biosensor based on the recombinant Staphylococcal protein A with an additional cysteine residue (SPA-Cys) used as a biorecognition component showed a good selectivity and sensitivity for the immunoglobulin detection. The developed biosensor with SPA-Cys-based bioselective element can also be used as a first step of immunosensor creation. The successful immobilization of SPA-Cys on the nanolayer gold sensor surface of the SPR spectrometer was performed.

Multiple immune deviations predictive for IVF failure as possible markers for IVIG therapy.

Recently we have shown that immune deviations (ID) may predict IVF failure. Benefit from IVIG therapy was observed in 115 women with repeated IVF failure according to proposed multiple ID that appeared unfavorable for implantation and live birth. Group of 123 women with repeated IVF failure without IVIG therapy was compared with former group.

Repeated cupping manipulation temporary decreases natural killer lymphocyte frequency, activity and cytotoxicity.

Objective: Elevated natural killer lymphocyte cytotoxicity (NKc) has been linked with reproductive problems in women. Here we evaluate the potential benefit of cupping therapy (CT) in reproduction-related immune responses.

Methods: This was a pilot clinical study.

[REDUCED RESPONSE OF NATURAL KILLER LYMPHOCYTES TO TOLL-LIKE RECEPTOR 3 STIMULATION IN CHILDREN WITH RECURRENT INFECTIONS.].

In this study, we examined the activation of natural killer(NK)-lymphocytes mediated by Toll-like receptor 3(TLR3), in the group of children with recurrent infections and the group of children with invasive bacterial infections. We examined level of CD69 (marker, of activation) expression on NK - lymphocytes after incubation with TLR3 ligand. There was a significant decrease in the level of an activation marker - 36,3±4,4% in the group of children with recurrent infections, compared with a control group of healthy children (56,5±4,9%) and the group of children with invasive bacterial infections (55,9±4,4%).

Accentuated hypo- and hyper-NK lymphocyte CD8 expression is a marker of NK subsets' misbalance and is predictive for reproductive failures.

Introduction: NK cells play critical yet poorly defined role in reproductive processes. Role of CD8αα expression on a part of peripheral blood NK lymphocyte population is not clear.

Patients And Methods: During isolated double-blinded clinical investigation we studied blood samples from 153 women with multiple reproductive failures undergoing IVF which were drawn 5-12d before embryo transfer procedure.

Favorable immune phenotype predicts successful implantation and pregnancy.

Immune markers that may predict IVF failure and successful implantation and pregnancy were studied. Favorable immune parameters were selected based on 90% of data of women who got pregnant and had uneventful pregnancy course and outcome in present IVF cycle. Immune phenotype and NK cell activity of peripheral blood of 123 women with multiple IVF failure were studied by flow cytometry.

[Effect of hypo- and hyper- accentuated NK cell activity on embryo implantation].

NK lymphocytes play an important role in implantation and during development in early pregnancy. Recently, we showed that the proportion of NK that expressed CD69 after incubation with K562 (CD69(stim)) cells reflected the NK population excitation potential. In the present study, we investigated the significance of NK activation levels in predicting the implantation outcome in 84 patients following IVF (in vitro fertilization).

Peripheral blood natural killer cells activation status determined by CD69 upregulation predicts implantation outcome in IVF.

Problem: NK lymphocytes play critical yet poorly defined role in implantation and during development in early pregnancy.

Methods Of Study: Recently, we showed that the proportion of NK that expressed CD69⁺ after incubation with K562 (CD69(stim)) cells reflected the NK population excitation potential. In the present study, we investigate the significance of NK activation levels in predicting embryo implantation.

[Qualitative analysis of accented CD158a receptor expression in NK-lymphocytes in women with reproductive failures].

Blood from 91 women that undergoing IVF cycle was analysed for NK cells CD158a expression using a monoclonal antibody and FACScan flow cytometer and CellQuest software (BD Bioscience, San Jose, USA). Patients were separated on 3 groups according implantation and pregnancy results in actual IVF cycle. 53 patients not became pregnant (IVF failure group F), 24 became pregnant after IVF with subsequent successful pregnancy (Pregnancy succes group PS) and 13 became pregnant with subsequently pregnancy failure (Pregnancy failure group PF).

Preimplantation factor inhibits circulating natural killer cell cytotoxicity and reduces CD69 expression: implications for recurrent pregnancy loss therapy.

Embryo-secreted preimplantation factor (PIF) is necessary for, and its concentration correlates with, embryo development in humans by promoting implantation and trophoblast invasion. Synthetic PIF (sPIF) modulates systemic immunity and is effective in autoimmune disease models. sPIF binds monocytes and activated T and B cells, leading to immune tolerance without suppression.

Measurement of NK activity in whole blood by the CD69 up-regulation after co-incubation with K562, comparison with NK cytotoxicity assays and CD107a degranulation assay.

In present study human peripheral blood NK cell activation after co-incubation with K569 cell line was investigated by CD69 expression. NK lytic activity was studied by two different assays: TDA (2,2':6',2″-terpyridine-6,6″-dicarboxylate) release assay (TRA) and flow cytometry assay (FCA) that display two approach to cytotoxicity measurement. We also investigated NK cell degranulation activity by estimation of CD107a (LAMPa) expression.

Elevated NK cell cytotoxicity, CD158a expression in NK cells and activated T lymphocytes in peripheral blood of women with IVF failures.

Problem: The aim of this study was to evaluate the role of elevated natural killer cytotoxicity (NKc) in women with multiple implantation failures (IF) in vitro fertilization-embryo transfer (IVF-ET) cycles.

Methods Of Study: Seventy-nine antiphospholipid antibodies-negative women with IF including 33 women with elevated NKc were selected for investigation. K-562 cell line was used to evaluate NKc.

[Anti-idiotypic antibodies as a physiological mechanism of inhibition of cofactor-independent antiphospolipid antibody formation].

Antiphospholipid antibodies (APA) and anti-idiotypic antiphospholipid antibodies (AiAPA) were studied in 148 women subjected to IVF. APA (IgG aCL, aPS,) levels in serum have been defined. Sera IgG fraction was also examined for the presence of AiAPA by three different methods: 1) binding of AiAPA with mouse monoclonal cofactor-independent APA (mAPA) immobilized on plate; 2) AiAPA neutralization of human affinity isolated APA and 3) mAPA binding with phospholipids.

[Cofactor-antiphospholipid antibodies relationship in patients with systemic lupus erythematosus and infertility women].

High frequency of detecting antiphospholipid antibodies (APA) is typical for patients with systemic autoimmune diseases as well as infertility women irresponsive to IVF treatment. Antiphospholipid antibodies cause thrombotic complications in patients with systemic autoimmune diseases contrary to antiphospholipid antibodies with in-vitro fertilization taken place. The authors have compared specificity and cofactor-antiphospholipid antibodies relationship in patient with systemic lupus erythematosus, infertility women irresponsive to IVF treatment and patients with chronic virus hepatitis.