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. 2019 Apr;26(4):532-542.
doi: 10.1177/1933719118777638. Epub 2018 Jun 18.

Trophoblast-Derived CXCL16 Decreased Granzyme B Production of Decidual γδ T Cells and Promoted Bcl-xL Expression of Trophoblasts

Deng-Xuan Fan  1 , Wen-Jie Zhou  1 , Li-Ping Jin  2 , Ming-Qing Li  1 , Xiang-Hong Xu  2 , Cong-Jian Xu  1   3   4
Affiliations

Trophoblast-Derived CXCL16 Decreased Granzyme B Production of Decidual γδ T Cells and Promoted Bcl-xL Expression of Trophoblasts

Deng-Xuan Fan et al. Reprod Sci. 2019 Apr.

Abstract

Background: Decidual γδ T cells are known to regulate the function of trophoblasts at the maternal-fetal interface; however, little is known about the molecular mechanisms of cross talk between trophoblast cells and decidual γδ T cells.

Methods: Expression of chemokine C-X-C motif ligand 6 (CXCL16) and its receptor CXCR6 was evaluated in first-trimester human villus and decidual tissues by immunohistochemistry. γδ T cells were isolated from first-trimester human deciduae and cocultured with JEG3 trophoblast cells. Cell proliferation and apoptosis-related molecules, together with cytotoxicity factor and cytokine production, were measured by flow cytometry analysis.

Results: Expression of CXCL16 and CXCR6 was reduced at the maternal-fetal interface in patients who experienced unexplained recurrent spontaneous abortion as compared to healthy pregnancy women. With the administration of pregnancy-related hormones or coculture with JEG3 cells, CXCR6 expression was upregulated on decidual γδ T cells. CXCL16 derived from JEG3 cells caused a decrease in granzyme B production of decidual γδ T cells. In addition, decidual γδ T cells educated by JEG3-derived CXCL16 upregulated the expression of Bcl-xL in JEG3 cells.

Conclusion: This study suggested that the CXCL16/CXCR6 axis may contribute to maintaining normal pregnancy by reducing the secretion of cytotoxic factor granzyme B of decidual γδ T cells and promoting the expression of antiapoptotic marker Bcl-xL of trophoblasts.

Keywords: CXCL16; CXCR6; decidual γδ T cells; trophoblasts; unexplained recurrent spontaneous abortion.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Reduced expression of CXCL16 and CXCR6 was found in villi and decidua of URSA patients. Immunohistochemistry analysis of the expression and localization of CXCL16 (A) and CXCR6 (B) was performed in villi from 10 women in the first trimester of pregnancy and 10 URSA patients. Representative images (100× and 400×) were shown. C, CXCL16 protein expression in the culture of JEG3 cells was analyzed by ELISA. Data were mean ± SEM from 5 independent experiments. *P < .05; ***P < .001. ELISA indicates enzyme-linked immunosorbent assay; NS, nonsignificant; SEM, standard error of the mean; URSA, unexplained recurrent spontaneous abortion.
Figure 2.
Figure 2.
Expression of CXCR6 was induced in decidual γδ T cells cocultured with JEG3 or treated by pregnancy-related hormones. A, Coculture with JEG3 trophoblast cells for 48 hours, CXCR6+ decidual γδ T cells were determined by flow cytometric analysis. B, A t test was performed for the statistical significance of percentage of CXCR6+ decidual γδ T cells between control cells and treated cells. C, After treatment with estrogen, progesterone, and human chorionic gonadotropin for 72 hours, respectively, CXCR6+ γδ T cells were determined by FCM. A t test was performed for significance testing. Data were mean ± SEM from 3 independent experiments. ***P < .001. FCM indicates flow cytometry; SEM, standard error of the mean.
Figure 3.
Figure 3.
JEG3 trophoblast cells promoted the cell viability and proliferation of decidual γδ T cells. After coculture with JEG3 and/or treatment with rhCXCL16 or CXCL16 neutralizing antibody for 48 hours, respectively, the cell viability (A) and proliferation (B) of decidual γδ T cells were detected. A t test was performed for significance testing. Data were mean ± SEM from 3 independent experiments. ***P < .001. NS indicates nonsignificant; SEM, standard error of the mean.
Figure 4.
Figure 4.
JEG3 trophoblast cells downregulated the secretion of granzyme B from decidual γδ T cells dependent of CXCL16. A, Coculture with JEG3 and/or treatment with rhCXCL16 or CXCL16 neutralizing antibody for 48 hours, respectively, the secretion of granzyme B, IFN-γ, and perforin of γδ T cells was analyzed by FCM. B, A t test was performed for significance testing. Data were mean ± SEM from 3 independent experiments. *P < .05; **P < .01; ***P < .001. FCM indicates flow cytometry; IFN-γ, interferon γ; NS, nonsignificant; SEM, standard error of the mean.
Figure 5.
Figure 5.
JEG3 trophoblast cells induced the cytokine production in decidual γδ T cells. Coculture with JEG3 and/or treatment with rhCXCL16 or CXCL16 neutralizing antibody for 48 hours, respectively, the expression of cytokines (IL-4, IL-5, IL-10, TGF-β, TNF-α, and IL-17A) was determined by FCM. A t test and 1-way analysis of variance were performed for significance testing. Data were mean ± SEM from 12 independent experiments. **P < .01; ***P < .001. FCM indicates flow cytometry; IL, interleukin; NS, nonsignificant; SEM, standard error of the mean; TGF-β, transforming growth factor β; TNF-α, tumor necrosis factor α.
Figure 6.
Figure 6.
Decidual γδ T cells educated by JEG3-derived CXCL16 upregulated Bcl-xL expression. A, A diagram of cell treatment and sample collection progresses was given. B, The expression of cell proliferation and apoptosis-related markers (ki-67, Fas, FasL, and Bcl-xL) was analyzed by FCM for JEG3. C-F A t test was performed for significance testing. Data were mean ± SEM from 3 independent experiments. **P < .01; ***P < .001. FCM indicates flow cytometry; NS, nonsignificant; SEM, standard error of the mean.
Figure 7.
Figure 7.
The proposed mechanism of cross talk between decidual γδ T cells and trophoblast cells mediated by CXCL16/CXCR6 during pregnancy. Treatment with pregnancy-related hormones or coculture with JEG3 trophoblast cells, CXCR6 expression was upregulated on decidual γδ T cells. CXCL16 derived from JEG3 trophoblast cells caused a decrease in granzyme B production of decidual γδ T cells. Decidual γδ T cells educated by JEG3-derived CXCL16 upregulated the expression of Bcl-xL in JEG3 trophoblast cells. Maintaining appropriate trophoblast growth is important for a normal healthy pregnancy.
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References

    1. Fujiwara H, Higuchi T, Yamada S, et al. Human extravillous trophoblasts express laeverin, a novel protein that belongs to membrane-bound gluzincin metallopeptidases. Biochem Biophys Res Commun. 2004;313(4):962–968. - PubMed
    1. Ji L, Brkic J, Liu M, Fu G, Peng C, Wang YL. Placental trophoblast cell differentiation: physiological regulation and pathological relevance to preeclampsia. Mol Aspects Med. 2013;34(5):981–1023. - PubMed
    1. Baczyk D, Dunk C, Huppertz B, et al. Bi-potential behaviour of cytotrophoblasts in first trimester chorionic villi. Placenta. 2006;27(4-5):367–374. - PubMed
    1. Huang Y, Zhu XY, Du MR, Li DJ. Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy. J Immunol. 2008;180(4):2367–2375. - PubMed
    1. Huang Y, Zhu XY, Du MR, Wu X, Wang MY, Li DJ. Chemokine CXCL16, a scavenger receptor, induces proliferation and invasion of first-trimester human trophoblast cells in an autocrine manner. Hum Reprod. 2006;21(4):1083–1091. - PubMed

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