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Pregnancy is a state where high and stage-dependent plasticity of the maternal immune system is necessary in order to equilibrate between immunosuppression of harmful responses towards the fetus and ability to fight infections. TCR γδ cells have been implicated in the responses in infectious diseases, in the regulation of immune responses, and in tissue homeostasis and repair. The variety of functions makes γδ T cells a particularly interesting population during pregnancy. In this study, we investigated the proportion, phenotype and TCR γ and δ repertoires of γδ T cells at the maternal⁻fetal interface and in the blood of pregnant women using FACS, immunohistochemistry and spectratyping. We found an enrichment of activated and terminally differentiated pro-inflammatory γδ T-cell effectors with specific location in the human decidua during early pregnancy, while no significant changes in their counterparts in the blood of pregnant women were observed. Our spectratyping data revealed polyclonal CDR3 repertoires of the δ1, δ2 and δ3 chains and γ2, γ3, γ4 and γ5 chains and oligoclonal and highly restricted CDR3γ9 repertoire of γδ T cells in the decidua and blood of pregnant women. Early pregnancy induces recruitment of differentiated pro-inflammatory γδ T-cell effectors with diverse TCR repertoires at the maternal⁻fetal interface.

Aim: To investigate human granulosa luteinized cells as a local source of atrial natriuretic peptide (ANP) and its relationship to cell viability, development of preovulatory follicles and fertilization rate of oocytes., Material and Methods: Indirect immunofluorescent technique, proANP kit, 4'6-Diamidino-2-phenylindole (DAPI)-DNA staining and Caspase-3 activity assay were used to examine localization patterns, concentrations in follicular fluids (FFl) and antiapoptotic role of ANP, respectively., Results: ANP is expressed on granulosa cells with membrane, submembrane and specific granular cytoplasmic localization. Significant differences in the mean concentrations of ANP in FFl and in the mean levels of apoptotic human granulosa luteinized cells (GLC) were found between women with high and low immunoreactive ANP expression (P < 0.05). Concentrations of ANP correlated inversely and significantly with the percentage of apoptotic GLC isolated from the same women (r = -0.2645; P < 0.05). Enhanced in vitro production of ANP by granulosa cells (after sodium nitroproside and 2Bu-cGMP treatment), as well as after treatment with human recombinant ANP was associated with significant suppression of Caspase-3 activity compared to control cells (P < 0.05). Concentrations of ANP in FFl correlated positively and significantly with the number of punctured follicles and oocytes retrieved (r = 0.44716, r = 0.57854; P < 0.05) and with the fertilization rate of oocytes in vitro (r = 0.59773; P < 0.05)., Conclusion: Human granulosa luteinized cells are a local source of ANP in the preovulatory human follicle, which acts as a survival factor through cGMP signaling, and its levels in follicular fluids are positively associated with the number of punctured follicles, isolated oocytes and their fertilizability in vitro., (© 2011 The Authors. Journal of Obstetrics and Gynaecology Research © 2011 Japan Society of Obstetrics and Gynecology.)

Objective: Our aim was to examine the expression patterns of ANP, the rate of apoptosis bcl-2 and p53 expression and caspase-3 activity and progesterone (P) production in porcine granulosa cells (pGCs) stimulated in vitro for luteinization and after treatment with leptin., Methods: Freshly isolated prepubertal pGCs were cultured as monolayers for 24 h, subsequently FSH was supplemented for 24 h, and finally LH was added to a part of the cells for 24 h to induce luteinization. The effect of leptin on in vitro luteinized pGCs was tested by the addition of 10 ng/ml human recombinant leptin (hrL) 24 h after LH administration. Indirect immunofluorescence for ANP, bcl-2 and p53 expression was used, P production was assayed by direct enzyme immunoassay (EIA) and colorimetric AcDEVD-Pna assay for caspase-3 activity was applied., Results: Stimulation of prepubertal pGCs with FSH resulted in a moderate expression of ANP and elevation in P production. When FSH treatment was followed by LH, the pronounced expression of ANP was observed in all cells. Suppressive effect of FSH and LH on p53 expression and caspase-3 activity with parallel increase in bcl-2 expression and increased P production was observed. The treatment of in vitro luteinized (FSH/LH-stimulated) pGCs with leptin did not influence the expression of ANP. However, in FSH/LH plus leptin treated cells the concomitant increase in bcl-2 expression and parallel inhibitory effect on p53 expression and caspase-3 activity was noted, compared to control cells without any significant increase in P production., Conclusion: The present data demonstrated the positive relationship between ANP expression and P production in pGCs stimulated for luteinization in vitro by FSH and LH, as well as their antiapoptotic role mediated presumably by cGMP accumulation in the luteinized pGCs. A direct anti-apoptotic effect of leptin on in vitro luteinized pGCs, without any significant modulation of P production, was documented.