Caselli Elisabetta, Bortolotti Daria, Marci Roberto, Rotola Antonella, Gentili Valentina, Soffritti Irene, D'Accolti Maria, Lo Monte Giuseppe, Sicolo Mariangela, Barao Isabel, Di Luca Dario, Rizzo Roberta
Section of Microbiology and Medical Genetics, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
School of Medicine, University of Geneva, Geneva, Switzerland.
Front Microbiol. 2017 Dec 15;8:2525. doi: 10.3389/fmicb.2017.02525. eCollection 2017.
We have recently reported the presence of Human herpesvirus-6A (HHV-6A) DNA in the 43% of endometrial epithelial cells from primary idiopathic infertile women, with no positivity in fertile women. To investigate the possible effect of HHV-6A infection in endometrial (e)NK cells functions, we examined activating/inhibitory receptors expressed by eNK cells and the corresponding ligands on endometrial cells during HHV-6A infection. Endometrial biopsies and uterine flushing samples during the secretory phase were obtained from 20 idiopathic infertile women and twenty fertile women. HHV-6A infection of endometrial epithelial cells was analyzed by Real-Time PCR, immunofluorescence and flow cytometry. eNKs receptors and endometrial ligands expression were evaluated by immunofluorescence and flow cytometry. We observed the presence of HHV-6A infection (DNA, protein) of endometrial epithelial cells in the 40% of idiopathic infertile women. The eNK from all the subgroups expressed high levels of NKG2D and NKG2A receptors. Functional studies showed that NKG2D activating receptor and FasL are involved in the acquired cytotoxic function of eNK cells during HHV-6A infection of endometrial epithelial cells. In the presence of HHV-6A infection, eNK cells increased expression of CCR2, CXCR3 and CX3CR1 chemokine receptors ( = 0.01) and endometrial epithelial cells up-modulated the corresponding ligands: MCP1 (Monocyte chemotactic protein 1, CCL2), IP-10 (Interferon gamma-induced protein 10, CXCL10) and Eotaxin-3 (CCL26). Our results, for the first time, showed the implication of eNK cells in controlling HHV-6A endometrial infection and clarify the mechanisms that might be implicated in female idiopathic infertility.
我们最近报道，在原发性特发性不孕女性的43%子宫内膜上皮细胞中存在人疱疹病毒6A（HHV-6A）DNA，而在有生育能力的女性中未检测到阳性。为了研究HHV-6A感染对子宫内膜（e）自然杀伤细胞功能的可能影响，我们检测了HHV-6A感染期间eNK细胞表达的激活/抑制受体以及子宫内膜细胞上的相应配体。在分泌期从20名特发性不孕女性和20名有生育能力的女性中获取子宫内膜活检组织和子宫冲洗样本。通过实时聚合酶链反应、免疫荧光和流式细胞术分析子宫内膜上皮细胞的HHV-6A感染情况。通过免疫荧光和流式细胞术评估eNK细胞受体和子宫内膜配体的表达。我们观察到40%的特发性不孕女性的子宫内膜上皮细胞存在HHV-6A感染（DNA、蛋白质）。所有亚组的eNK细胞均高表达NKG2D和NKG2A受体。功能研究表明，在子宫内膜上皮细胞感染HHV-6A期间，NKG2D激活受体和FasL参与了eNK细胞获得性细胞毒性功能。在存在HHV-6A感染的情况下，eNK细胞增加了趋化因子受体CCR2、CXCR3和CX3CR1的表达（P = 0.01），子宫内膜上皮细胞上调了相应的配体：单核细胞趋化蛋白1（MCP1，CCL2）、干扰素γ诱导蛋白10（IP-10，CXCL10）和嗜酸性粒细胞趋化因子3（CCL26）。我们的结果首次表明eNK细胞在控制HHV-6A子宫内膜感染中的作用，并阐明了可能与女性特发性不孕有关的机制。
