PMID- 28673654 OWN - NLM STAT- MEDLINE DCOM- 20180406 LR - 20180409 IS - 1879-0542 (Electronic) IS - 0165-2478 (Linking) VI - 188 DP - 2017 Aug TI - Competent antigen-presenting cells are generated from the long-term culture of splenocytes with granulocyte-macrophage colony-stimulating factor. PG - 96-107 LID - S0165-2478(17)30083-4 [pii] LID - 10.1016/j.imlet.2017.06.010 [doi] AB - Dendritic cells (DCs) are routinely produced from the culture of mouse bone marrow (BM) with granulocyte-macrophage colony-stimulating factor (GM-CSF) within a period of 10days. Although splenic extramedullary myelopoiesis was suggested to occur under the influence of GM-CSF, the hematopoietic outcome of splenic culture with GM-CSF has not been scrutinized. We have cultured mouse splenocytes with GM-CSF for an extended period of time, where we discovered that the CD11b(+)CD11c(+) cells began to proliferate prominently after 10days and their number increased until the 4th week of the culture. In parallel experiments, FMS-like tyrosine kinase 3 (FLT3) and its ligand, FLT3L, were not found to influence the culture of splenocytes. Like DCs in the culture of BM with GM-CSF, a distinct population of CD11b(+)CD11c(+)MHC II(hi) cells was readily identified as DCs in the long-term culture of splenocytes. After being isolated and plated overnight the CD11b(+)CD11c(+)MHC II(hi) cells exhibited non-adherent dendritic morphology, while the other CD11b(+)CD11c(+) cells became adherent. Besides, these CD11b(+)CD11c(+)MHC II(hi) cells possessed relatively weak endocytic and phagocytic abilities but displayed strong antigen-presenting capacities, revealing DC-like characteristics; in contrast, the other CD11b(+)CD11c(+) cells showed strong endocytosis and phagocytosis of antigens but were poor at antigen presentation, indicating macrophage-like traits. Therefore, we demonstrated that phenotypically as well as functionally genuine DCs are generated in the long-term culture of splenocytes with GM-CSF. CI - Copyright (c) 2017 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved. FAU - Ryu, Seul Hye AU - Ryu SH AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - Na, Hye Young AU - Na HY AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - Sohn, Moah AU - Sohn M AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - Choi, Wanho AU - Choi W AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - In, Hyunju AU - In H AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - Shin, Hyun Soo AU - Shin HS AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. FAU - Choi, Jae-Hoon AU - Choi JH AD - Department of Life Science, College of Natural Sciences, Research Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea. FAU - Park, Chae Gyu AU - Park CG AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. Electronic address: ChaeGyu@yuhs.ac. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170630 PL - Netherlands TA - Immunol Lett JT - Immunology letters JID - 7910006 RN - 0 (Biomarkers) RN - 0 (Histocompatibility Antigens Class II) RN - 0 (Membrane Proteins) RN - 0 (flt3 ligand protein) RN - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor) RN - EC 2.7.10.1 (Flt3 protein, mouse) RN - EC 2.7.10.1 (fms-Like Tyrosine Kinase 3) SB - IM MH - Animals MH - Antigen Presentation/immunology MH - Antigen-Presenting Cells/cytology/*drug effects/*immunology/metabolism MH - Biomarkers MH - Bone Marrow Cells MH - Cell Culture Techniques MH - Cells, Cultured MH - Dendritic Cells/immunology/metabolism MH - Granulocyte-Macrophage Colony-Stimulating Factor/*pharmacology MH - Histocompatibility Antigens Class II/immunology MH - Immunophenotyping MH - Membrane Proteins/metabolism MH - Mice MH - Mice, Transgenic MH - Signal Transduction MH - Spleen/*cytology/*immunology/metabolism MH - T-Lymphocyte Subsets/immunology/metabolism MH - fms-Like Tyrosine Kinase 3/metabolism OTO - NOTNLM OT - Antigen-presenting cells OT - Dendritic cells OT - GM-CSF OT - Hematopoiesis OT - Spleen EDAT- 2017/07/05 06:00 MHDA- 2018/04/07 06:00 CRDT- 2017/07/05 06:00 PHST- 2017/02/16 00:00 [received] PHST- 2017/05/29 00:00 [revised] PHST- 2017/06/26 00:00 [accepted] PHST- 2017/07/05 06:00 [pubmed] PHST- 2018/04/07 06:00 [medline] PHST- 2017/07/05 06:00 [entrez] AID - S0165-2478(17)30083-4 [pii] AID - 10.1016/j.imlet.2017.06.010 [doi] PST - ppublish SO - Immunol Lett. 2017 Aug;188:96-107. doi: 10.1016/j.imlet.2017.06.010. Epub 2017 Jun 30.