title: CD4(+)CD25(+)Foxp3(+)IFNγ(+) Treg are immunosuppressive in vitro and increase with intensity of the alloresponse in pretransplant MLC.
abstract: IFNγ-producing CD3(+)CD4(+)CD25(+)Foxp3(+) induced Treg are more frequently detectable in patients with good than in patients with impaired long-term kidney graft function. We investigated the in-vitro function of separated CD3(+)CD4(+)CD25(+)Foxp3(+)IFNγ(+) PBL that were induced by phorbol-12-myristate-13-acetate(PMA)/Ionomycin or alloantigenic stimulation. Additionally, we studied iTreg induction and cell proliferation in MLC with pretransplant obtained PBL. CD4(+)CD25(+)IFNγ(+) PBL separated from PMA/Ionomycin-stimulated PBL of healthy controls inhibited secondary cell cultures of autologous PBL. Furthermore, CD4(+)CD25(+)IFNγ(+) PBL separated from primary MLC and added to secondary MLC suppressed allogeneic T-cell activation in secondary MLC unspecifically, irrespective of the stimulator cell. However, the strongest suppression was observed in specific MLC. Patients with poor long-term graft outcome were able to form IFNγ(+) iTreg in pretransplant MLC. Eight patients with a serum creatinine level ranging from 0.9 to 14 mg/dl 18-29 years posttransplant were studied. In MLC with pretransplant obtained recipient and donor cells, strong IFNγ(+) iTreg (p=0.007) and strong blast induction (p=0.047) were associated with impaired long-term graft outcome. Long-term graft outcome was not associated with cell proliferation and iTreg induction in unspecific MLC with third-party cells as stimulator. The data indicate that patients with impaired long-term graft outcome are able to form high numbers of IFNγ(+) iTreg in specific pretransplant MLC. Quantity of induced IFNγ(+) iTreg depends on the strength of the alloresponse and both parameters are inversely associated with long-term graft outcome.
authors.affiliation: Department of Transplantation-Immunology, Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, 69120 Heidelberg, Germany. Volker.Daniel@med.uni-heidelberg.de
authors.forename: Volker
authors.initials: V
authors.lastname: Daniel
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Sadeghi
authors.forename: Haihao
authors.initials: H
authors.lastname: Wang
authors.forename: Gerhard
authors.initials: G
authors.lastname: Opelz
mesh_terms: D015704:CD4 Antigens / Q000378:metabolism; D002478:Cells, Cultured; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D007111:Immunity, Cellular; D007165:Immunosuppression Therapy; D007371:Interferon-gamma / Q000378:metabolism*; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D007519:Isoantigens / Q000276:immunology; D016030:Kidney Transplantation / Q000276:immunology*; D018655:Lymphocyte Count; D007959:Lymphocyte Culture Test, Mixed; D059035:Perioperative Period; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: D015704:CD4 Antigens; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053645:Interleukin-2 Receptor alpha Subunit; D007519:Isoantigens; D007371:Interferon-gamma