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The Cross-talking between Decidual γδ T Cells and Trophoblasts in Human Early Pregnancy

Author: FanDengXuan
Tutor: JinLiPing
School: Fudan University
Course: Obstetrics and Gynaecology
Keywords: Gestation - Maternal interface γδT cells Trophoblastic IL-10 CXCL16 Proliferation Apoptosis Attack
CLC: R339.22
Type: Master's thesis
Year: 2011
Downloads: 36
Quote: 0
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Abstract


Physiological pregnancy is similar to the allograft. Until delivery as allograft embryo in the mother's memory live to reflect maternal immune tolerance embryos; maternal immune rejection of embryonic lead to pregnancy failure. Reveal the exact mechanism - maternal immune tolerance, will have a great significance on the prevention and treatment of human spontaneous abortion and other pregnancy-related diseases, and to promote the role of transplant immunology and tumor immunology research. - Maternal interface nourish cells, endometrial stromal cells, decidual glandular epithelial cells and immune cells. Over the years people from different perspectives - maternal interface of biological events, to elaborate - maternal immune regulation mechanism. - Maternal interface there is a large number of lymphocytes, and nourishes the cultured cells in close contact. Thus, decidual lymphocytes, decidual cells and nourish cells among the inevitable existence of close functional regulation of participation - maternal immune regulation. Although a limited number of decidual γδT cells, but its function can not be ignored. Our group preliminary work as the starting point of the chemokine receptor, has been elucidated - maternal interface trophoblastic through CXCL16/CXCR6 the specific chemokine raise γδT cells to the mother - fetal interface. This study further resolved γδT cells, the regulatory role of the mother - fetal interface, through the establishment of human decidua γδT cells and trophoblast cells co-cultured system, intended to clarify the mechanism of mutual adjustment between the decidua γδT cells and trophoblast. The purpose of the first part of the first trimester the mother - fetal interface γδT cell phenotype: analysis of human first trimester maternal the γδT cell ratio changes and phenotypic characteristics, and intracellular cytokine expression. Methods: normal early pregnant women of childbearing age normal non-pregnant women in peripheral blood and decidua (endometrium) organization, the proportion of γδT cells using flow cytometry analysis of peripheral blood and decidual immune cells; using immunomagnetic beads cell sorting isolated and purified decidual γδT cells by flow cytometry decidual γδT cells, CD4 and CD8 and intracellular cytokine expression (IL-2, TNF-α, IFN-γ, IL-4 of IL- 10, TGF-β1). Results: normal early pregnant women in the peripheral blood and decidual local γδT cell number than normal non-pregnant women were significantly increased, and decidual γδT cells was significantly higher than the proportion of peripheral blood. These results suggest that an increase in the number of pregnant women γδT cells to maintain the normal pregnancy. Decidual γδT cells, isolated and purified γδT cells in more than 95% purity. Decidual γδT cells are CD4-and CD8-double negative cells. The γδT cells high levels of expression of TGF-β and IL-10, low-level expression of TNF-α and IFN-γ, and almost no expression of IL-2 and IL-4. Conclusion: decidual γδT cells produce large amounts of regulatory cytokines, Th2-type immune microenvironment advantage in decidual local formation, thus contributing to tolerance - maternal formation. The second part of early pregnancy decidual γδT cells for the purpose of regulation of trophoblast cell biology: explore early pregnancy decidual γδT cell regulation of trophoblast cell biology. Methods: decidua isolated decidual immune cells, decidual γδT cells obtained bead sorting. Collect the villi primary isolation and culture of early pregnancy trophoblastic. ΓδT cells and trophoblast cells co-culture system for 48h, BrdU cell proliferation ELISA kit to detect trophoblastic proliferation; Annexin Ⅴ / PI double staining kit, flow cytometry analysis of apoptosis of trophoblastic situation; γδT cells trophoblastic invasion through the establishment of a transwell system. Result: decidual γδT cells induced trophoblast proliferation and invasion, and IL-10 neutralizing antibody significantly inhibited this induction of TGF-β neutralizing antibodies did not; recombinant human IL-10 treatment trophoblastic significantly promote trophoblast proliferation and invasion, and recombinant human TGF-β has no effect. Decidual γδT cells inhibit trophoblast apoptosis, and IL-10 neutralizing antibodies significantly antagonized the inhibitory effect, but does not have this role of TGF-β neutralizing antibodies; trophoblast cells treated with recombinant human IL-10, then significantly inhibit trophoblast apoptosis, recombinant human TGF-β does not have this effect. Conclusion: decidual γδT cells through the secretion of IL-10 to promote trophoblast proliferation and invasion, and inhibition of trophoblast cell apoptosis. Therefore, γδT cells can improve the trophoblastic biological behavior conducive to the formation of the placenta. The third part of early pregnancy trophoblastic decidual γδT cells of the biological function of the regulatory role Objective: To investigate the first trimester trophoblast cell function regulation on decidual γδT cells: trophoblast cells and γδT cell co-culture system based platform add or not add CXCL16 neutralizing antibodies and human recombinant CXCL16 cytokines by flow cytometry analysis of the regulatory role of trophoblast cells and CXCL16/CXCR6 γδT cell proliferation and analysis of trophoblastic on γδT cells granzyme B expression regulation role. Using ELISA analysis of cytokine TGF-β and IL-10 in the culture supernatant. Results: trophoblast cells can promote the proliferation of γδT cells, CXCL16 neutralizing antibody significantly inhibited this role; recombinant human CXCL16 separate processing decidual γδT cells significantly promote the the decidual γδT cell proliferation. Trophoblast cells or human recombinant CXCL16 can be down-regulation of the expression of γδT cells GrB; the γδT cells anti-human CXCL16 neutralizing antibodies could antagonize drop mediation GrB expression. The trophoblastic promotion γδT cells produce TGF-β and IL-10. Conclusion: early pregnancy, trophoblast cells by secreting CXCL16 promote the decidual γδT cell proliferation, and produce TGF-β and IL-10 increase in the formation of a virtuous cycle conducive to pregnancy; trophoblast cells through decidual γδT cells secrete CXCL16 down granzyme B level, down-regulation of γδT cells of embryonic tissue damage, in order to maintain a normal pregnancy.

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CLC: > Medicine, health > Basic Medical > Human Physiology > Reproductive (sex ) and physiological > Female genital physiology
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