Regulation of CD4⁺FOXP3⁺ T cells by CCL20/CCR6 axis in early unexplained recurrent miscarriage patients.
Expression and function of CCR6/CCL20 in CD4(+)FOXP3(+) regulatory T cells (Tregs) was investigated in unexplained recurrent miscarriage (URM) patients. Flow cytometry, reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, western blots, and Transwell migration assays were used to analyze the expression and function of regulatory T cells in peripheral blood (PB) and decidual samples of women with URM and of healthy controls. Proportions of CD4(+)FOXP3(+) T cells and CCR6(+)CD4(+)FOXP3(+) T cells were lower in URM patients than in healthy controls for both PB lymphocytes and decidual samples (P < 0.05). Expression levels of FOXP3 and CCR6 mRNA were lower in URM patients than in control subjects for PB and decidual samples (P < 0.05). CCL20 protein levels were lower in URM patients than in controls (P < 0.05). An effect of Treg migration was significantly blocked (by 89.13%) using a neutralizing anti-CCL20 antibody in vitro. Furthermore, CCL20-stimulated Tregs exhibited a 3.21-fold increase in migration and this was blocked using a neutralizing anti-CCL20 antibody. IL-10 concentration in culture supernatants of CD4(+)CD25(+)CD127(dim/-) Tregs of URM patients was significantly lower than that in controls. Anti-CCL20 antibody inhibited IL-10 and IL-4 expression but increased IFN-r and IL-17 levels when there was cell-cell contact between PB CD4(+)CD25(+) T cells and CD4(+)CD25(-) T cells. No difference was detected when cell-cell contact was prevented by a semi-permeable Transwell membrane. CCL20-CCR6 could drive immune activity of CD4(+)FOXP3(+) Tregs, followed by their migration to the feto-maternal microenvironment. These results elucidated the mechanism by which Tregs exert this suppressive effect.
Department of Obstetrics and Gynecology, Renji Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, China.
X X
XX
Zhang
Department of Obstetrics and Gynecology, Renji Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, China.
X M
XM
Kang
Department of Obstetrics and Gynecology, Renji Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, China zamzkh0526@126.com.
A M
AM
Zhao
D000026:Abortion, Habitual / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D054418:Chemokine CCL20 / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis / Q000276:immunology; D020381:Interleukin-17 / Q000096:biosynthesis / Q000276:immunology; D015847:Interleukin-4 / Q000096:biosynthesis / Q000276:immunology; D011247:Pregnancy; D054399:Receptors, CCR6 / Q000096:biosynthesis / Q000235:genetics*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000378:metabolism* / Q000473:pathology
C108355:CCL20 protein, human; C106311:CCR6 protein, human; D054418:Chemokine CCL20; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; C508609:IL10 protein, human; C508717:IL17A protein, human; D020381:Interleukin-17; D054399:Receptors, CCR6; D016753:Interleukin-10; D015847:Interleukin-4