Early Pregnancy Human Decidua is Enriched with Activated, Fully Differentiated and Pro-Inflammatory Gamma/Delta T Cells with Diverse TCR Repertoires.

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.

Atrial natriuretic peptide is an antiapoptotic factor for human granulosa luteinized cells with impact on the results of COH/IVF in women undergoing IVF program.

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.

Nitric oxide stimulates the production of atrial natriuretic peptide and progesterone by human granulosa luteinized cells with an antiapoptotic effect.

OBJECTIVE: To study the effect of nitric oxide (NO) on atrial natriuretic peptide (ANP) and progesterone (P) production by human granulose luteinized cells (GLC) in vitro and to elucidate their role on the survival of cultured cells. METHODS: Human GLCs were cultured in HAM's F10/10% FCS as monolayers for 24 h. Subsequently GLCs were treated for 24 h with 0.5 mM Sodium Nitroprusside (SNP, NO donor) and 0.5 mM Aminoglutethimide (AG , P450scc inhibitor). The levels of ANP and P were measured in supernatants of cultured cells by proANP(1-98) kit and RIA, respectively. Caspase-3 activity was determined by Ac-DEVD-pNA as substrate. RESULTS: The production of ANP and P was increased by NO as compared to control cells (p<0.05). AG diminished the production of P compared to SNP (p<0.05). The caspase-3 activity was significantly lower in SNP treated cells (p<0.05) and increased significantly after AG treatment compared to control cells (p<0.05). CONCLUSION: NO generated by SNP in human GLCs culture stimulated the production of ANP and P. The higher levels of ANP and P were closely related to significantly lower caspase-3 activity thus showing the role of ANP, P and NO on the survival of preovulatory human follicle.