PMID- 24239650 OWN - NLM STAT- MEDLINE DCOM- 20141103 LR - 20240213 IS - 1523-6536 (Electronic) IS - 1083-8791 (Print) IS - 1083-8791 (Linking) VI - 20 IP - 2 DP - 2014 Feb TI - IL-12hi rapamycin-conditioned dendritic cells mediate IFN-gamma-dependent apoptosis of alloreactive CD4+ T cells in vitro and reduce lethal graft-versus-host disease. PG - 192-201 LID - S1083-8791(13)00526-0 [pii] LID - 10.1016/j.bbmt.2013.11.007 [doi] AB - Rapamycin (RAPA) inhibits the mechanistic target of rapamycin (mTOR), a crucial immune system regulator. Dendritic cells (DC) generated in RAPA (RAPA-DC) enrich for CD4(+) forkhead box p3 (FoxP3(+)) regulatory T cells and induce T cell apoptosis by an unknown mechanism. RAPA-DC also promote experimental allograft survival, yet paradoxically secrete increased IL-12, crucial for the generation of IFN-gamma(+) CD4(+) T cells. However, IFN-gamma is pro-apoptotic and IL-12-driven IFN-gamma inhibits experimental graft-versus-host disease (GVHD). We hypothesized that IL-12(hi) RAPA-DC would facilitate IFN-gamma-mediated apoptosis of alloreactive T cells and, unlike control (CTR)-DC, would reduce lethal GVHD. Following LPS stimulation, RAPA-DC exhibited decreased MHCII and co-stimulatory molecules and contained a significant population of CD86(lo) IL-12(hi) cells. Consistent with our hypothesis, both unstimulated and LPS-stimulated RAPA-DC enhanced alloreactive CD4(+) T cell apoptosis in culture. Augmented T cell apoptosis was ablated by IFN-gamma neutralization or using T cells lacking the IFN-gamma receptor, and it was associated with increased expression of Fas and cleaved caspase 8. DC production or responses to IFN-gamma were not important to increased apoptotic functions of RAPA-DC. LPS-stimulated IL-12p40(-/-) RAPA-DC induced lower levels of T cell apoptosis in culture, which was further decreased with addition of anti-IFN-gamma. Finally, whereas CTR-DC accelerated mortality from GVHD, LPS-treated RAPA-DC significantly prolonged host survival. In conclusion, increased apoptosis of allogeneic CD4(+) T cells induced by LPS-stimulated IL-12(hi) RAPA-DC is mediated in vitro through IFN-gamma and in part by increased IL-12 expression. Enhanced production of IL-12, the predominant inducer of IFN-gamma by immune cells, is a probable mechanism underlying the capacity of LPS-treated RAPA-DC to reduce GVHD. CI - Copyright (c) 2014 American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved. FAU - Stenger, Elizabeth O AU - Stenger EO AD - Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Division of Pediatric Hematology/Oncology, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania. FAU - Rosborough, Brian R AU - Rosborough BR AD - Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. FAU - Mathews, Lisa R AU - Mathews LR AD - Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. FAU - Ma, Huihui AU - Ma H AD - Division of Hematology/Oncology, Department of Medicine, Hematologic Malignancies Program, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania. FAU - Mapara, Markus Y AU - Mapara MY AD - Division of Hematology/Oncology, Department of Medicine, Hematologic Malignancies Program, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania. FAU - Thomson, Angus W AU - Thomson AW AD - Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. FAU - Turnquist, Heth R AU - Turnquist HR AD - Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. Electronic address: het5@pitt.edu. LA - eng GR - R01HL93716/HL/NHLBI NIH HHS/United States GR - R01 AI060994/AI/NIAID NIH HHS/United States GR - T32 AI074490/AI/NIAID NIH HHS/United States GR - R00 HL097155/HL/NHLBI NIH HHS/United States GR - NIH T32 AI74490/AI/NIAID NIH HHS/United States GR - K99/R00 HL97155/HL/NHLBI NIH HHS/United States GR - R01AI60994/AI/NIAID NIH HHS/United States GR - R01 GM063569/GM/NIGMS NIH HHS/United States GR - R01 HL093716/HL/NHLBI NIH HHS/United States GR - R01GM63569/GM/NIGMS NIH HHS/United States GR - K99 HL097155/HL/NHLBI NIH HHS/United States GR - R01AI67541/AI/NIAID NIH HHS/United States GR - R01 AI067541/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20131112 PL - United States TA - Biol Blood Marrow Transplant JT - Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation JID - 9600628 RN - 0 (Antibiotics, Antineoplastic) RN - 187348-17-0 (Interleukin-12) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - Animals MH - Antibiotics, Antineoplastic/immunology/*therapeutic use MH - Apoptosis/*immunology MH - CD4-Positive T-Lymphocytes/*immunology MH - Cell Differentiation MH - Cell Proliferation MH - Dendritic Cells/*immunology MH - Female MH - Graft vs Host Disease/*immunology MH - Interleukin-12/*immunology MH - Male MH - Mice MH - Mice, Inbred BALB C MH - Mice, Inbred C57BL MH - Sirolimus/immunology/*therapeutic use PMC - PMC3964782 MID - NIHMS541189 OTO - NOTNLM OT - Apoptosis OT - Dendritic cells OT - Graft-versus-host disease OT - IFN-gamma OT - Rapamycin OT - T cells COIS- Conflict of interest statement: A.W.T. is co-inventor of US patents that concern generation of dendritic cells to promote transplant tolerance. EDAT- 2013/11/19 06:00 MHDA- 2014/11/05 06:00 PMCR- 2015/02/01 CRDT- 2013/11/19 06:00 PHST- 2013/07/07 00:00 [received] PHST- 2013/11/06 00:00 [accepted] PHST- 2013/11/19 06:00 [entrez] PHST- 2013/11/19 06:00 [pubmed] PHST- 2014/11/05 06:00 [medline] PHST- 2015/02/01 00:00 [pmc-release] AID - S1083-8791(13)00526-0 [pii] AID - 10.1016/j.bbmt.2013.11.007 [doi] PST - ppublish SO - Biol Blood Marrow Transplant. 2014 Feb;20(2):192-201. doi: 10.1016/j.bbmt.2013.11.007. Epub 2013 Nov 12.