title: CD4+ CD25+ regulatory T cells in human pregnancy: development of a Treg-MLC-ELISPOT suppression assay and indications of paternal specific Tregs.
abstract: The current study was aimed at developing a one-way mixed leucocyte culture-enzyme-linked immunospot (MLC-ELISPOT) assay for the study of CD4(+) CD25(+) regulatory T (T(reg)) cells and applying this method in the study of antifetal immune reactions during human pregnancy. Twenty-one pregnant women and the corresponding fathers-to-be, and 10 non-pregnant control women and men, participated in the study. CD4(+) CD25(+) cells were isolated from peripheral blood mononuclear cells (PBMC) by immunomagnetic selection. Maternal/control PBMC were stimulated with paternal or unrelated PBMC in MLC. Secretion of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) from responder cells, with or without the presence of autologous T(reg) cells, was analysed by ELISPOT. PBMC from pregnant women showed increased secretion of IL-4 compared to controls. In pregnant and non-pregnant controls, T(reg) cells suppressed IFN-gamma reactivity against paternal and unrelated alloantigens. Interestingly, T(reg) cells suppressed IL-4 secretion against paternal but not unrelated alloantigens during pregnancy. We have successfully developed a model for studying T(reg) cells in antifetal cytokine reactions during pregnancy. Results indicate that T(reg) cells contribute to strict regulation of both T helper type 1-like and type 2-like antifetal immune reactions. Interestingly, T helper type 2-like cells specific to unrelated alloantigens are able to escape the suppression of T(reg) cells, which would allow for IL-4, alongside CD4(+) CD25(+) T(reg) cells, to control potentially detrimental IFN-gamma reactions during pregnancy.
authors.affiliation: Unit for Autoimmunity and Immunoregulation, Faculty of Health Sciences, Department of Molecular and Clinical Medicine, Linköping University, Linköping, Sweden. jenmj@imk.liu.se
authors.forename: Jenny
authors.initials: J
authors.lastname: Mjösberg
authors.forename: Göran
authors.initials: G
authors.lastname: Berg
authors.forename: Jan
authors.initials: J
authors.lastname: Ernerudh
authors.forename: Christina
authors.initials: C
authors.lastname: Ekerfelt
mesh_terms: D000293:Adolescent; D000328:Adult; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005333:Fetus / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods; D051858:Forkhead Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D007108:Immune Tolerance; D018189:Immunomagnetic Separation / Q000379:methods; D007371:Interferon-gamma / Q000096:biosynthesis; D053645:Interleukin-2 Receptor alpha Subunit / Q000032:analysis*; D007519:Isoantigens / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008875:Middle Aged; D011247:Pregnancy / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology
chemical_list: C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053645:Interleukin-2 Receptor alpha Subunit; D007519:Isoantigens; D012333:RNA, Messenger; D007371:Interferon-gamma