Changes of apoptosis regulation in the endometrium of infertile women with tubal factor and endometriosis undergoing in vitro fertilization treatment.

OBJECTIVE: To establish the relationship between the endometrial apoptosis parameters and endometrial receptivity in infertile women undergoing IVF treatment. METHODS: 73 women with tubal infertility, 27 infertile women with endometriosis and 13 healthy fertile women (control group) were recruited into the study. 53 women with tubal factor of infertility and 17 patients with endometriosis later entered in the IVF protocol. Samples of endometrial tissue were used as the material for investigation. Endometrium was collected on the 20-24 days of non-conceptual cycle, using the Pipell suction curette. XIAP, PTEN and HSP27 mRNAs expression in the endometrial tissue was assessed by real time RT-PCR. RESULTS: In women with tubal infertility the high level of XIAP, HSP27 and PTEN mRNAs expression in the endometrium was found. In infertile women with endometriosis the increase of XIAP and HSP27 mRNAs expression was noted. Success of the IVF outcome in women with tubal infertility was associated with maximal level of PTEN synthesis and in endometriosis group the pregnancy achievement after IVF treatment was noted in women with lower expression of XIAP mRNA. CONCLUSION: The high level of pro-apoptotic factors synthesis in the endometrium during window of implantation is associated with the readiness of endometrium to implantation.

Mechanisms regulating invasiveness and growth of endometriosis lesions in rat experimental model and in humans.

OBJECTIVE: To compare the expression of MMP-2, TIMP-2, and TGFbeta2 mRNA in experimental and human endometriotic lesions and to assess the possibility of its cytokine regulation. DESIGN: Experimental laboratory study. SETTING: Medical center. ANIMALS AND PATIENT(S): Thirty female Wistar rats, 17 women with endometriosis, 11 healthy women. INTERVENTION(S): Uterine transplants were attached to rat peritoneum via the surgical autotransplantation technique. The collection of endometriotic implants at 7, 14, and 21 days postsurgery and laparoscopic collection of peritoneal fluid, ectopic, and matched eutopic endometrium from women with endometriosis were performed. MAIN OUTCOME MEASURE(S): MMP-2, TIMP-2, TGFbeta2 mRNA expression in endometrium was assessed by real-time reverse-transcription polymerase chain reaction. RESULT(S): In rats, the increase of MMP-2 and decrease of TIMP-2 mRNA expression was noted at the 7th day, and an increase of TGFbeta2 mRNA expression was seen at the 14th day postsurgery. In humans, elevation of TIMP-2 mRNA expression in eutopic endometrium and of MMP-2, TGFbeta2 mRNA expression in ectopic endometrium was observed. Autologous peritoneal fluid stimulated MMP-2 mRNA expression in eutopic endometrium of women with endometriosis. Cytokines derived from ectopic lesions mononuclear cells increased TGFbeta2 mRNA expression in endometrium of healthy women. CONCLUSION(S): Supposedly MMP-TIMP balance is important in promoting endometriotic tissue invasion and TGFbeta2 in regulating ectopic endometrium growth.

Changes in the T-helper cytokine profile and in lymphocyte activation at the systemic and local levels in women with endometriosis.

OBJECTIVE: To estimate regulatory cytokine synthesis and lymphocyte activation in the peripheral blood and endometrial tissue of patients with endometriosis. DESIGN: Controlled clinical study. SETTING: Medical center. PATIENT(S): Fifteen women with laparoscopically diagnosed endometriosis; 20 gynecologically healthy women with previously documented fertility (control group). INTERVENTION(S): Peripheral venous blood sampling; laparoscopic collection of ectopic and matched eutopic endometrium. MAIN OUTCOME MEASURE(S): Messenger RNA (mRNA) for interleukin (IL)-2, IL-4, and IL-10 expression in peripheral and endometrium lymphocytes was assessed by real-time reverse transcriptase polymerase chain reaction; intracellular synthesis of these cytokines and lymphocyte phenotype profile were established by flow cytometry. RESULT(S): Both mRNA expression and intracellular synthesis of IL-4 and IL-10 were sharply increased in peripheral lymphocytes. The same changes were observed for IL-4 in ectopic endometrium of women with endometriosis. Simultaneously, elevation of the amount of pan-B cells, CD20+CD5+ B-1 cells, and activated HLA-DR+CD20+ B lymphocytes was observed in endometriosis lesions. Only an enhanced amount of B lymphocytes was seen in eutopic endometrium. CONCLUSION(S): Endometriosis development is accompanied by the activation of a T-helper type 2 immune response at the systemic and local levels. Our results support the hypothesis regarding the autoimmune nature of endometriosis and can explain the high level of autoantibody production in patients with endometriosis.

Effect of alvitili on immunological parameters in often ailing children.

The problem of the rehabilitation of often ailing children is not yet elucidated despite of a substantial amount of research done in this field. It is acknowledged that the impairment of immune reactivity is a pathogenetic foundation of often and prolonged respiratory diseases in children. In this connection, the aim of the study was to evaluate the effect of alvitili on the main immunologic parameters in children who get ill frequently. Alvitili, produced by Solvay Pharma (France), is one of the promising vitamin preparations that contain all necessary water-soluble and oil-soluble vitamins. Thirty children 3-8 years of age were selected for the study: 15 children were included in the group of often ailing children and 15 healthy children made the control group. Immunological examination was carried out using standard methods before and after administration of alvitili at a dose of 5 ml 3 times a day for 3 weeks. Alvitili normalized originally decreased levels of T lymphocytes and T helpers, enhanced expression of early activation markers on the surface of T lymphocytes and T helpers, normalized the number of HLA DR(+) T cells and the level of MIF production. The drug stimulated IgM synthesis, expression of FcgammaRIII on the surface of neutrophils and normalized the IgG level. The study showed that alvitili is an effective remedy in the rehabilitation of ill children, since it compensates a vitamin deficiency that is frequently seen in often-ailing children, and it normalizes various immunological parameters.

Dual effect of cerebrolysin in children with attention deficit syndrome with hyperactivity: neuroprotection and immunomodulation.

Children with attention deficit syndrome with hyperactivity (ADSH) are characterised by attention and motor deficiencies and obvious immune disorders, which manifest as recurrent acute viral respiratory tract infections and changes in immunological parameters. It was established that the structure of deviations in the spectrum of elements in children with ADSH has characteristic features. The atomic emission analysis of cerebrolysin detected an advantageous combination of trace elements with neuroactive and antioxidant properties. The effect of cerebrolysin on immunological parameters was studied in vitro and in vivo. Cerebrolysin was administered in a dose of 1 ml per 10 kg of weight intramuscularly during 1 month. The administration of cerebrolysin resulted in a simultaneous normalization of neurological and immune disorders and in a reduction in the illness rate. In in vitro experiments cerebrolysin enhanced the expression of activation markers (HLA DR, CD25) by CD45(+)CD14(-) lymphocytes, particularly by CD4(+) cells. In vivo it led to the normalization of the numbers of CD4(+), CD19(+), CD16(+) and CD56(+) cells and of the level of serum IgG and IgA. Cerebrolysin normalized the expression of HLA DR molecules on the surface of CD8(+) cells and increased the amount of CD11b(+) lymphocytes. At the same time, it did not affect the level of CD95 molecule expression.

Local immunological markers of different rate of growth of uterine myoma.

The aim of our work was to establish the peculiarities of phenotype profile and functional activity of peritoneal immunocompetent cells of women with different rate of myoma growth. Forty three women with uterine myoma (main group) and 16 healthy women (control group) were recruited for study. According to the growth rate and tumor's size the main group was divided into two clinical subgroups: (1) women with stable small size of uterine myoma (23 patients), (2) women with large size rapidly growing myoma (20 patients). The expression of CD markers on the surface of peritoneal fluid (PF) lymphocytes and bactericidal and adhesive activity of macrophages were studied. It was found that in women with stable small myomas both lymphocytes and macrophages in PF were activated. In women with rapidly growing large myomas cellular reactions seemed to be inhibited and the impairment of maturation and differentiation of lymphocytes was observed. It can be suggested that local immune response is directly connected with the rate of myoma growth.

Local Epidermal Growth Factor Production in Women with Endometriosis.

The aim of the study was to elucidate the role of epidermal growth factor (EGF) in pathogenesis of endometriosis at different stages of disease. The level of EGF in peritoneal fluid (PF) of fertile and infertile women with endometriosis and in pooled supernatants of 24-h cultures of peritoneal macrophages was assessed by ELISA method. It was found that in fertile women with early stages of disease EGF level in PF did not differ from that in control group, but at the advanced stages of endometriosis the significant elevation of EGF concentration in PF was observed. The spontaneous production of EGF by peritoneal macrophages in this group was diminished. It might be suggested that elevation of EGF concentration in PF in this group is the result of secretory activity of endometriosis lesions and macrophages didn't regulate the local EGF production in fertile women. EGF level in PF of infertile women was increased only in women with minimal manifestations of endometriosis but not in women with advanced stages of the disease. Simultaneously the increase of EGF production by peritoneal macrophages of infertile women with advanced stages of endometriosis was shown. These facts give an evidence in favor of high level of macrophage activation and their participation in regulation of EGF level in peritoneal cavity in infertile women with endometriosis.

A Study of Peritoneal Immunocompetent Cells in External Genital Endometriosis.

The aim of the present investigation was to study the peculiarities of peritoneal lymphoid cell activation at the local level during external endometriosis and the possible role of secretory products of peritoneal macrophages in this process. Immunocompetent cells of peritoneal fluid from 16 women with endometriosis and 14 healthy women were studied by two-color flow cytometry method. It was found that endometriosis was associated with high level of expression of CD25 and HLA DR molecules and diminishment of Fas expression by CD3(+) subset of peritoneal lymphocytes. Experimentally it was established that supernatant of 24-h culture of peritoneal macrophages from women with endometriosis induced the expression of CD25 marker by CD4(+) but not CD8(+) cells and decreased Fas expression by both cell subsets in donor lymphocytes. It can be supposed that altered secretory function of peritoneal macrophages in situ impair the apoptosis of activated clones of T lymphocytes. Observed effect of macrophage secretory products was not associated with TNF-alpha production by peritoneal macrophages in the process of their cultivation.