Expression of retinoid-related orphan receptor (ROR)γt on NK22 cells in the peripheral blood and uterine endometrium of women with unexplained recurrent pregnancy loss and unexplained infertility.

AIM: Recently, NK22 cells, a subset of interleukin (IL)-22-producing natural killer (NK) cells, were identified. We have previously reported the higher percentage of NK22 cells in women suffering recurrent pregnancy loss (RPL). Moreover, we have also reported lower expression of NKp46, a kind of natural cytotoxicity receptor (NCR), on NK cells and the changes of NK cell producing cytokines in women who experience RPL. NK22 cells express NCRs, such as NKp44 or NKp46. Retinoid-related orphan receptor γt (RORγt) is known as a regulator of NK22 cells; however, in NK22 cells of peripheral blood (PB) and the uterine endometrium (UE), the relationship between NCRs and RORγt is unclear. We investigate RORγt expression NK22 cells in the PB and UE of women with unexplained infertility (uI) or unexplained RPL (uRPL). METHODS: Lymphocytes were extracted from PB and UE, derived from women with uI or uRPL. Expression of RORγt and NCRs in NK cells and NK cell-produced cytokines were analyzed by flow cytometry. RESULTS: CD56+ /NKp46+ /RORγt+ cells were positively correlated with CD56+ /IL-22+ cells in both PB and UE. CD56bright /NKp46bright /RORγt+ cells were significantly higher in uRPL than in uI, and endometrial CD56bright /NKp46bright /RORγt+ cells were positively correlated with PB. In UE, CD56bright /RORγt+ cells were negatively correlated with CD56bright /interferon-γ+ and CD56bright /tumor necrosis factor-α+ cells of uRPL. CONCLUSION: RORγt may be associated with NK22 cells in reproduction. Particularly, higher expression of RORγt may be associated with elevated NK22 cells in uRPL.

Expression of Natural Cytotoxicity Receptors on and Intracellular Cytokine Production by NK Cells in Women with Gestational Diabetes Mellitus.

PROBLEM: To determine the role of peripheral blood NK (pNK) cells in putative etiology of gestational diabetes, the expression of surface markers on pNK cells and the percentage of cytokine-producing pNK cells in women at 12 weeks of pregnancy with gestational diabetes mellitus (GDM) were studied. METHOD OF STUDY: Multicolor flow cytometry was used to analyze the expression of NK cell surface receptors (CD16, NKp46, and NKp30) and intracellular cytokines (IFN-γ, TNF-α, TGF-ß, and VEGF) in pNK cells (CD56(dim) and CD56(bright) ) at 12 weeks of pregnancy with GDM (n = 7) and non-GDM (n = 28). RESULTS: CD56(bright) /CD16(-) NK and CD56(bright) /NKp46(+) NK cell percentage were significantly lower in GDM women than that in non-GDM women. IFN-γ- and TNF-α-producing CD56(+) cells, respectively, were significantly high, while TGF-ß- and VEGF-producing CD56(+) cells and CD56(bright) cells, respectively, were significantly low in GDM women. CONCLUSIONS: Women with GDM possibly have abnormal NK cell function for the expression of surface receptors and cytokine production.

NK22 Cells in the Uterine Mid-Secretory Endometrium and Peripheral Blood of Women with Recurrent Pregnancy Loss and Unexplained Infertility.

PROBLEM: We aimed to investigate natural killer 22 (NK22) cells in the peripheral blood and the uterine endometrium of women with unexplained recurrent pregnancy loss (URPL) and unexplained infertility (UI). METHOD OF STUDY: Peripheral blood and endometrial samples were collected from women with URPL (n = 43) and UI (n = 38). Intracellular cytokine production, such as IL-22, IFN-γ and TNF-α, and the expression of NKp46 on NK cells were analyzed by three-color flow cytometry. RESULTS: The percentages of endometrial CD56(+) /IL-22(+) and CD56(dim) /IL-22(+) cells in women with URPL were significantly higher than those of UI (P < 0.05, respectively). In addition, the percentage of CD56(bright) /IL-22(+) cells in women with RPL was negatively correlated with those of CD56(bright) /IFN-γ(+) and CD56(bright) /TNF-α(+) in both peripheral blood and endometrial NK cells. This was not seen in women with UI. The percentage of CD56(bright) /IL-22(+) cells was negatively correlated with CD56(bright) /NKp46 expressing NK cells in peripheral blood. CONCLUSION: Endometrial NK22 cells are differently regulated in women with URPL and UI. Women with URPL have higher level of NK22 cells with a potential to induce NK2 shift than women with UI.

NK cell abnormality and its treatment in women with reproductive failures such as recurrent pregnancy loss, implantation failures, preeclampsia, and pelvic endometriosis.

The regulation of uterine and peripheral blood natural killer (NK) cells has been associated with problems related to reproductive immunology such as recurrent pregnancy loss (RPL), implantation failure or preeclampsia. NKp46, one of the natural cytotoxicity receptors (NCRs), is a unique marker that functions in NK cell cytotoxicity and cytokine production. Expression of NKp46 on NK cells is lower in women with recurrent pregnancy loss and pregnancy-induced hypertension. Moreover, expression of NKp46 on peritoneal fluid NK cells is lower in women with pelvic endometriosis. Therefore, evaluation of NKp46 on peripheral blood NK cells may provide a means of screening for reproductive abnormalities. Recently, a new type of NK cell, the NK22 cell, has been reported. This cell may be a regulator not only of the mucosal barrier but also of reproduction. For women with RPL showing abnormal uterine and/or peripheral blood NK cells, both intravenous immunoglobulin treatment and intralipid treatment have been reported. The effects of these treatments are still controversial, and further studies are needed in order to clarify their true impact. The present review examines variations in the expression of NCRs on NK cells, the participation of NK22 cells in reproduction, and the possible use of intravenous immunoglobulin or intralipid treatment for women with recurrent pregnancy loss and NK cell abnormality.

Expression of natural cytotoxicity receptors on peritoneal fluid natural killer cell and cytokine production by peritoneal fluid natural killer cell in women with endometriosis.

PROBLEM: To investigate the relationship between the expression of natural cytotoxicity receptors (NCRs) on peritoneal fluid (PF) natural killer (NK) (pfNK) cells and cytokine production by pfNK cells in women with endometriosis. METHOD OF STUDY: Peritoneal fluid was collected from women with endometriosis undergoing laparoscopic surgery (n = 21) and controls without endometriosis (n = 28). The expression of NK cell surface antigens such as CD16 and NCRs (NKp46, NKp44 and NKp30) on pfNK cells, and cytokines production by pfNK cells [tumor necrosis factor (TNF)-α, IFN-γ, IL-4, IL-10, GM-CSF and transforming growth factor (TGF)-ß1] were measured using multicolor flow cytometry. RESULTS: The percentages of CD56(+)/NKp46(+) cells and CD56(dim) /NKp46(+) cells in severe endometriosis group were significantly lower than that in controls. TNF-α and IFN-γ production by pfNK cells in severe endometriosis group was significantly higher than those in controls. CONCLUSION: The differential expression of NKp46, TNF-α, and IFN-γ on pfNK cells in women with severe endometriosis may allow the proliferation and angiogenesis of endometriotic cells.

Role of NKp46 expression in cytokine production by CD56-positive NK cells in the peripheral blood and the uterine endometrium.

PROBLEM: To investigate the role of natural cytotoxicity receptor, NKp46 expression in cytokine-producing NK cells. METHOD OF STUDY: CD56(+) /NKp46(+) NK cells from the peripheral blood and the uterine endometrium were magnetically separated. IFN-γ, TNF-α, IL-4, IL-10, and TGF-ß(1) expressions on NK cells were investigated using multicolor flow cytometry. RESULTS: Peripheral blood and uterine endometrial NK cells were grouped into 4 subpopulations based upon the degree of CD56 and NKp46 expressions. NKp46 expression was associated with higher frequency of cytokine-producing NK cells, including CD56(dim) NK cells. The percentage of TNF-α(+) and IL-10(+) NK cells per total CD56(+) /NKp46(+) NK cells in the uterine endometrium showed a significant correlation with those of the peripheral blood in all subpopulations, but that of IFN-γ(+) , IL-4(+,) and TGF-ß(1) (+) NK cells showed partial correlation. CONCLUSION: Expression of NKp46 is involved in cytokine production of CD56(+) NK cells in the peripheral blood and the uterine endometrium.

Successful pregnancy and delivery after transfer of a single blastocyst derived from a vitrified mature human oocyte.

OBJECTIVE: To describe the first case of delivery after the transfer of a single blastocyst derived from a vitrified mature human oocyte. DESIGN: Case report. SETTING: Private assisted reproduction clinic. PATIENT(S): A normal 31-year-old woman. INTERVENTION(S): An unsuccessful attempt was made to extract sperm from the patient's nonobstructive azoospermic husband by testicular sperm extraction. MAIN OUTCOME MEASURE(S): Transfer of single blastocyst derived from vitrified human oocyte and donor sperm. RESULT(S): A healthy male neonate weighing 3000 g was born. CONCLUSION(S): Vitrification is a useful method of preserving mature human oocytes and has the advantage of time-effectiveness, simplicity, cost reduction, and no need for devices such as programmable freezers.

Successful pregnancy and delivery following combined treatment of in vitro maturation (IVM) and testicular sperm extraction (TESE): a case report.

PURPOSE: To report the efficacy of the combined treatment of in vitro maturation (IVM) and testicular sperm extraction (TESE). METHODS: A couple in which the wife had polycystic ovarian syndrome and the husband had severe oligozoospermia. Oocytes were cultured in vitro for maturation followed by oocytes pickup with natural cycle, and TESE was undergone for husband. Matured oocytes were fertilized by intracytoplasmic sperm injection, and two embryos were transferred to wife's uterine. RESULTS: This case was achieved during pregnancy and delivery of a healthy female infant. CONCLUSIONS: The combined treatment of IVM and TESE was effective for this couple's specific infertility factors.

Freezing of human immature oocytes using cryoloops with Taxol in the vitrification solution.

PURPOSE: In human frozen immature oocytes, there has been little successful delivery. We examined the feasibility of vitrification solution including Taxol, cytoskeltal stabilizer. METHODS: We set four experimental groups that immature oocytes has cumulus cells or not, or including Taxol or not in the vitrification solution. Frozen-thawed oocytes have been performed IVM, ICSI, and IVC. RESULTS: There were no significant differences in survival, maturation, and fertilization rate, respectively. However, in the group enveloped by cumulus cells and including Taxol in the vitrification solution, one embryo was developed to blastocyst. CONCLUSIONS: Our results showed that using vitrification solution with Taxol proved so effective.

Physical and mental development of children after in vitro fertilization and embryo transfer.

Objective: To evaluate the physical and mental development of children after in vitro fertilization (IVF) and frozen embryo transfer (FET). Methods: Between July 1995 and November 2003, 506 patients delivered 658 babies after IVF and ET treatment at our clinic (intracytoplasmic sperm injection (ICSI), 418; conventional IVF, (C-IVF) 125; FET, 115). A survey of the physical and mental developmental of the children was conducted by mailing questionnaires to parents. Comparisons were made between three treatment procedures, and development of singleton, twin and triplet delivery. Results: The response rate was 73.4% (483/658) for 324 children born after ICSI, 78 born after C-IVF, and 81 born after FET. The height and weight of assisted reproductive technology (ART) children at birth were significantly lower than that of naturally conceived babies (ART children: natural delivery; 46.8 cm, 49.0 cm and 2524 g, 3040 g, respectively). However, there was no significant difference between the singletons alone and naturally conceived children irrespective of the ART method. In addition, mental development was the same between singletons and naturally conceived children. The ART group tended to delay body development such as 'holding their head up', 'sitting up', 'crawl' to moving growth in multiple births. Conclusion: The physical and mental development of twins or triplets was significantly more delayed than that of naturally conceived babies, but had improved to a similar extent of the singletons after the age of 6 months. (Reprod Med Biol 2004; 3: 63-67).

Two-step consecutive transfer of early embryos and blastocysts.

Aim: To evaluate and confirm the merit of two consecutive transfer attempts of early embryos and blastocysts. Methods: A total of 685 patients underwent routine in vitro fertilization (IVF) cycles. The study population consisted of three groups: (i) Group 1, early stage embryos were transferred (460 patients; 567 cycles); (ii) group 2, blastocysts were transferred (88 patients; 105 cycles); and (iii) group 3, a two-step (consecutive) transfer was performed on 137 women (141 cycles). Specifically, a standard embryo transfer was performed on day 3, together with a two-step (consecutive) transfer of blastocysts. After the early embryo transfer, an extended culture of supernumerary embryos was conducted, followed by a second transfer of blastocyst(s). Results: No significant differences were found in the three groups with regards to either pregnancy or implantation rates for groups 1, 2, or 3; pregnancy: 34.6, 29.9, and 33.6%, respectively; implantation: 18.6, 15.9, and 15.1%, respectively. The miscarriage rate for each group was also not significant; 20.4, 30.8 and 28.6% for groups 1, 2 and 3, respectively. The multiple pregnancy rate of the three groups was 30.4, 30.8 and 35.7%, respectively, and these were not found to be significant results. Conclusion: No significant difference was found between the three groups with regards to the pregnancy, implantation, multiple pregnancy and miscarriage rates. The multiple pregnancy rate was highest in the two-step (consecutive) transfer group. This difference was not regarded to be highly significant. (Reprod Med Biol 2003; 2: 133-137).