PMID- 38094300 OWN - NLM STAT- MEDLINE DCOM- 20231216 LR - 20240315 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 14 DP - 2023 TI - Identification of dendritic cell precursor from the CD11c(+) cells expressing high levels of MHC class II molecules in the culture of bone marrow with FLT3 ligand. PG - 1179981 LID - 10.3389/fimmu.2023.1179981 [doi] LID - 1179981 AB - Dendritic cells (DCs) are readily generated from the culture of mouse bone marrow (BM) treated with either granulocyte macrophage-colony stimulating factor (GM-CSF) or FMS-like tyrosine kinase 3 ligand (FLT3L). CD11c(+)MHCII(+) or CD11c(+)MHCII(hi) cells are routinely isolated from those BM cultures and generally used as in vitro-generated DCs for a variety of experiments and therapies. Here, we examined CD11c(+) cells in the BM culture with GM-CSF or FLT3L by staining with a monoclonal antibody 2A1 that is known to recognize mature or activated DCs. Most of the cells within the CD11c(+)MHCII(hi) DC gate were 2A1(+) in the BM culture with GM-CSF (GM-BM culture). In the BM culture with FLT3L (FL-BM culture), almost of all the CD11c(+)MHCII(hi) cells were within the classical DC2 (cDC2) gate. The analysis of FL-BM culture revealed that a majority of cDC2-gated CD11c(+)MHCII(hi) cells exhibited a 2A1(-)CD83(-)CD115(+)CX(3)CR1(+) phenotype, and the others consisted of 2A1(+)CD83(+)CD115(-)CX(3)CR1(-) and 2A1(-)CD83(-)CD115(-)CX(3)CR1(-) cells. According to the antigen uptake and presentation, morphologies, and gene expression profiles, 2A1(-)CD83(-)CD115(-)CX(3)CR1(-) cells were immature cDC2s and 2A1(+)CD83(+)CD115(-)CX(3)CR1(-) cells were mature cDC2s. Unexpectedly, however, 2A1(-)CD83(-)CD115(+)CX(3)CR1(+) cells, the most abundant cDC2-gated MHCII(hi) cell subset in FL-BM culture, were non-DCs. Adoptive cell transfer experiments in the FL-BM culture confirmed that the cDC2-gated MHCII(hi) non-DCs were precursors to cDC2s, i.e., MHCII(hi) pre-cDC2s. MHCII(hi) pre-cDC2s also expressed the higher level of DC-specific transcription factor Zbtb46 as similarly as immature cDC2s. Besides, MHCII(hi) pre-cDC2s were generated only from pre-cDCs and common DC progenitor (CDP) cells but not from monocytes and common monocyte progenitor (cMoP) cells, verifying that MHCII(hi) pre-cDC2s are close lineage to cDCs. All in all, our study identified and characterized a new cDC precursor, exhibiting a CD11c(+)MHCII(hi)CD115(+)CX(3)CR1(+) phenotype, in FL-BM culture. CI - Copyright (c) 2023 In, Park, Shin, Ryu, Sohn, Choi, Park, Hwang, Park, Che, Kim, Chu, Na and Park. FAU - In, Hyunju AU - In H AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Park, Ji Soo AU - Park JS AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Shin, Hyun Soo AU - Shin HS AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Ryu, Seul Hye AU - Ryu SH AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Heart-Immune-Brain Network Research Center, Department of Life Science, Ewha Womans University, Seoul, Republic of Korea. FAU - Sohn, Moah AU - Sohn M AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Choi, Wanho AU - Choi W AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Park, Sejung AU - Park S AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Hwang, Soomin AU - Hwang S AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Park, Jeyun AU - Park J AD - Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Che, Lihua AU - Che L AD - Brain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Kim, Tae-Gyun AU - Kim TG AD - Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Chu, Min Kyung AU - Chu MK AD - Department of Neurology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Na, Hye Young AU - Na HY AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Department of Neurology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea. FAU - Park, Chae Gyu AU - Park CG AD - Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. AD - Heart-Immune-Brain Network Research Center, Department of Life Science, Ewha Womans University, Seoul, Republic of Korea. AD - Laboratory of Dendritic Cell Immunology, The Good Capital Institute for Immunology, Seoul, Republic of Korea. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20231129 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (Histocompatibility Antigens Class II) RN - 0 (flt3 ligand protein) RN - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor) RN - 0 (CX3C Chemokine Receptor 1) RN - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases) SB - IM MH - Mice MH - Animals MH - *Histocompatibility Antigens Class II MH - *Bone Marrow MH - Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology/metabolism MH - CX3C Chemokine Receptor 1/metabolism MH - Bone Marrow Cells MH - Dendritic Cells MH - Cell Differentiation MH - Receptor Protein-Tyrosine Kinases/metabolism PMC - PMC10716454 OTO - NOTNLM OT - FLT3 ligand OT - GM-CSF OT - antigen presentation OT - bone marrow cells OT - cell differentiation OT - cultured cells OT - dendritic cells OT - precursor cells COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2023/12/14 06:42 MHDA- 2023/12/17 09:45 PMCR- 2023/01/01 CRDT- 2023/12/14 04:14 PHST- 2023/03/05 00:00 [received] PHST- 2023/11/08 00:00 [accepted] PHST- 2023/12/17 09:45 [medline] PHST- 2023/12/14 06:42 [pubmed] PHST- 2023/12/14 04:14 [entrez] PHST- 2023/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2023.1179981 [doi] PST - epublish SO - Front Immunol. 2023 Nov 29;14:1179981. doi: 10.3389/fimmu.2023.1179981. eCollection 2023.