Immunoregulatory effects of the Blastocyst Conditioned Media: focus on the control of the inflammatory response

GRASSO, ESTEBAN; FERNÁNDEZ, LAURA DEL CARMEN; RUHLMANN, CLAUDIO; PAPARINI, DANIEL; MARTINEZ, GUSTAVO; RAMHORST, ROSANNA; SOCZEWSKI, ELIZABETH; IRIGOYEN, MARCELA; PEREZ LEIRÓS, CLAUDIA

Ginebra

Congreso; 33rd Annual Meeting of the European Society of Human Reproduction and Embryology; 2017

European Society of Human Reproduction and Embryology

Immunoregulatory effects of the Blastocyst Conditioned Media: focus on the control of the inflammatory responseE. Grasso, E. Soczewski, D. Paparini, L. Fernández, C. M. Irigoyen, G. Martinez, C. Ruhlmann, Pérez Leirós and R. RamhorstIQUIBICEN-CONICET, Dpto Química Biológica, School of Sciences, University of Buenos Aires, Argentina.Fertilidad San Isidro, Buenos Aires, ArgentinaStudy question: Could the human Blastocyst Conditioned Media (BCM) control the initial inflammatory response during the peri-implantation period toward a tolerogenic one? Summary answer: Decidualized cell responded differentially to BCM, accordingly their quality, by reducing reticular stress-associated inflammation, recruiting regulatory T lymphocytes (Tregs) and allowing blastocyst-like spheroids invasion.What is known already: The decidualization program of human endometrial cells involves a physiological reticular stress (RE) and unfolded protein response (UPR), allowing them to expand their endoplasmic reticulum and change their secretome increasing the production of immunomodulators. This physiological RE and UPR response would allow the activation of one of the 3 RE sensing proteins, IRE1, which induces kinase / RNase TXNIP expression, and activates the inflammasome generating a sterile inflammatory response with production of IL-1. This sterile inflammatory response, associated to the implantation period, should be later controlled in favor of a tolerogenic microenvironment by maternal and blastocyst-derived factors. Study design, size, duration: Human endometrial stromal cell line (HESC) were decidualized or not with medroxiprogesterone+dbcAMP during 8 days, and used to evaluated RE and UPR. Then, HESC cells were treated with human Blastocyst-condition-media (BCM), previously classified accordingly their quality, for 24h. An in vitro implantation model based on co-culture of blastocyst-like spheroids (BLS) from Swan-71 trophoblast cells line over decidualized-HESC cells and a transwell migration system for Tregs recruitment were used to evaluate decidual functionality. Participants/materials, setting, methods: Mature oocytes were inseminated (conventional IVF or ICSI was applied according to male evaluation). After 3 days, embryos were transferred individually to drops of G2 plus-medium. At day 5, BCM were retrieved and embryos classified following Istanbul concensus. Maternal monuclear cells were obtianed by pheripheral blood and Tregs frequency quantified by intracellular Foxp3 staining and FACS analysis. Gene and protein expression was evaluated by PCR or FACS respectively. BLS invasion was evaluated by fluorescence microscopy.Main results and the role of chance: We observed a significantly increase in the expression of CXCL8, CXCL12 and IL 1 after the decidualization Since IL 1 can act as a ?double edge? mediator in early pregnancy, we evaluated BCM effect on IL-1 production. BCM reduced IL 1 intracellular production in decidualized cells (p