title: Immunomodulatory effects of Sanglifehrin A in the innate and acquired immune response of neonatal whole blood cells.
abstract: Previous investigations have shown the immunosuppressive activity of the immunophilin-binding macrolide Sanglifehrin A (SFA). In adults, SFA also exerts anti-inflammatory properties in lipopolysaccharide (LPS)-stimulated whole blood cultures. It was the aim of this study to investigate whether the unique properties of SFA are also present in the neonatal immune system, as neonates are susceptible to serious infection due to an immaturity of immune responses. We used a whole blood assay to investigate the impact of SFA on T-cell proliferation and secretion of T-cell cytokines upon Anti-CD3/Anti-CD28 costimulation. In addition, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) secretion was assessed in whole blood monocytes after LPS stimulation. Furthermore, the influence of SFA on LPS-induced signal transduction pathways, specifically the activity of p42/44, p38 and Ap-1, was assessed in neonatal PBMCs. Neonatal cord blood lymphocytes were found to have a diminished IL-4, IL-6, TNF-alpha and interferon-gamma (IFN-gamma) production upon Anti-CD3/Anti-CD28 costimulation compared to adult whole blood lymphocytes. In contrast, no significant differences were noted for either IL-2 production or proliferation of CD4+ cells. Upon addition of 1000nM SFA to neonatal whole blood cultures, a significant inhibition of both, T-cell cytokine secretion and proliferation was demonstrated. In line with data from adult whole blood cultures, SFA proved to be a strong inhibitor of LPS-induced expression of IL-6 and TNF-alpha in the neonatal whole blood system. In signal transduction studies of the LPS pathway, a potent inhibition of the protein kinase p42/44 was demonstrated. SFA was also shown to block nuclear translocation of the transcription factor Ap-1. SFA was proved to have inhibitory effects on innate and acquired immune response of neonatal whole blood cells. The protein kinase p42/44 and the transcription factor Ap-1 were demonstrated to be potential key molecules for the anti-inflammatory effect of SFA.
authors.affiliation: Childrens Hospital, University of Lübeck Medical School, Ratzeburger Allee 160, 23538 Lübeck, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Härtel
authors.forename: Jan
authors.initials: J
authors.lastname: Rupp
authors.forename: Peter
authors.initials: P
authors.lastname: Iblher
authors.forename: Alexander
authors.initials: A
authors.lastname: Puzik
authors.forename: Ines
authors.initials: I
authors.lastname: Osthues
authors.forename: Christian
authors.initials: C
authors.lastname: Schultz
authors.forename: Michael
authors.initials: M
authors.lastname: Müller-Steinhardt
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate; D007155:Immunologic Factors / Q000494:pharmacology; D007156:Immunologic Memory; D007231:Infant, Newborn; D007783:Lactones / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology; D013141:Spiro Compounds / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000187:drug effects; D015533:Transcriptional Activation / Q000187:drug effects / Q000276:immunology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D016207:Cytokines; D007155:Immunologic Factors; D007783:Lactones; D008070:Lipopolysaccharides; D013141:Spiro Compounds; D018808:Transcription Factor AP-1; C121329:sanglifehrin A; D019950:Mitogen-Activated Protein Kinase 1; D048051:p38 Mitogen-Activated Protein Kinases