title: Induction of regulatory CD4⁺ cells and prolongation of survival of fully allogeneic murine cardiac grafts by danazol.
abstract: Danazol, a modified testosterone, has been used to treat endometriosis and pretreatment before in vitro fertilization and embryo transfer, although its reproductive mechanisms remain unclear. We investigated the effect of danazol on alloimmune responses in murine heart transplantation. CBA male mice (H2(k)) that underwent transplantation of C57BL/6 (B6, H2(b)) hearts received danazol (0.4 and 4 mg/kg/d) by intraperitoneal injection from the day of transplantation to days 6. We performed an adoptive transfer study to determine regulatory cells as well as cell proliferation, cytokine, and flow cytometry assessments. Danazol-treated (4 mg/kg/d) CBA mice showed prolonged allograft survival (median survival time [MST], 63 days). Moreover, secondary CBA recipients of whole splenocytes and CD4(+) cells from primary danazol-treated (4 mg/kg/d) CBA recipients at 30 days after transplantation displayed prolonged allograft survival (MSTs, 29 and 60 days, respectively). Cell proliferation, interleukin (IL)-2, and interferon-γ were suppressed in danazol-treated mice, whereas IL-4 and IL-10 were up-regulated. Moreover, danazol directly suppressed alloproliferation in mixed leukocyte cultures. Flow cytometry studies showed an increased CD4(+)CD25(+)Foxp3(+) cell population among splenocytes from danazol-treated mice. In conclusion, danazol induced prolonged cardiac allograft survival and generation of regulatory CD4(+) cells.
authors.affiliation: Department of Cardiovascular Surgery, Juntendo University Hospital, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Uchiyama
authors.forename: X
authors.initials: X
authors.lastname: Jin
authors.forename: Q
authors.initials: Q
authors.lastname: Zhang
authors.forename: A
authors.initials: A
authors.lastname: Amano
authors.forename: T
authors.initials: T
authors.lastname: Watanabe
authors.forename: M
authors.initials: M
authors.lastname: Niimi
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D003613:Danazol / Q000008:administration & dosage / Q000494:pharmacology*; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006084:Graft Rejection / Q000276:immunology / Q000517:prevention & control*; D006085:Graft Survival / Q000187:drug effects*; D016027:Heart Transplantation / Q000276:immunology*; D006648:Histocompatibility; D007155:Immunologic Factors / Q000008:administration & dosage / Q000494:pharmacology*; D007274:Injections, Intraperitoneal; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000276:immunology / Q000637:transplantation; D013997:Time Factors
chemical_list: D016207:Cytokines; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; C508018:Il2ra protein, mouse; D007155:Immunologic Factors; D053645:Interleukin-2 Receptor alpha Subunit; D003613:Danazol