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
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+MHCIIhi cells are routinely isolated from those BM cultures and gen...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-11-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1179981/full |
| _version_ | 1797448616448622592 |
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| author | Hyunju In Hyunju In Ji Soo Park Ji Soo Park Hyun Soo Shin Hyun Soo Shin Seul Hye Ryu Seul Hye Ryu Seul Hye Ryu Moah Sohn Moah Sohn Wanho Choi Wanho Choi Sejung Park Sejung Park Soomin Hwang Soomin Hwang Jeyun Park Lihua Che Lihua Che Tae-Gyun Kim Min Kyung Chu Hye Young Na Hye Young Na Chae Gyu Park Chae Gyu Park Chae Gyu Park |
| author_facet | Hyunju In Hyunju In Ji Soo Park Ji Soo Park Hyun Soo Shin Hyun Soo Shin Seul Hye Ryu Seul Hye Ryu Seul Hye Ryu Moah Sohn Moah Sohn Wanho Choi Wanho Choi Sejung Park Sejung Park Soomin Hwang Soomin Hwang Jeyun Park Lihua Che Lihua Che Tae-Gyun Kim Min Kyung Chu Hye Young Na Hye Young Na Chae Gyu Park Chae Gyu Park Chae Gyu Park |
| author_sort | Hyunju In |
| collection | DOAJ |
| description | 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+MHCIIhi 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+MHCIIhi 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+MHCIIhi cells were within the classical DC2 (cDC2) gate. The analysis of FL-BM culture revealed that a majority of cDC2-gated CD11c+MHCIIhi cells exhibited a 2A1-CD83-CD115+CX3CR1+ phenotype, and the others consisted of 2A1+CD83+CD115-CX3CR1- and 2A1-CD83-CD115-CX3CR1- cells. According to the antigen uptake and presentation, morphologies, and gene expression profiles, 2A1-CD83-CD115-CX3CR1- cells were immature cDC2s and 2A1+CD83+CD115-CX3CR1- cells were mature cDC2s. Unexpectedly, however, 2A1-CD83-CD115+CX3CR1+ cells, the most abundant cDC2-gated MHCIIhi cell subset in FL-BM culture, were non-DCs. Adoptive cell transfer experiments in the FL-BM culture confirmed that the cDC2-gated MHCIIhi non-DCs were precursors to cDC2s, i.e., MHCIIhi pre-cDC2s. MHCIIhi pre-cDC2s also expressed the higher level of DC-specific transcription factor Zbtb46 as similarly as immature cDC2s. Besides, MHCIIhi 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 MHCIIhi pre-cDC2s are close lineage to cDCs. All in all, our study identified and characterized a new cDC precursor, exhibiting a CD11c+MHCIIhiCD115+CX3CR1+ phenotype, in FL-BM culture. |
| first_indexed | 2024-03-09T14:13:02Z |
| format | Article |
| id | doaj.art-b7ad875009f64abbb484eb956afe332e |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-09T14:13:02Z |
| publishDate | 2023-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-b7ad875009f64abbb484eb956afe332e2023-11-29T05:53:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-11-011410.3389/fimmu.2023.11799811179981Identification of dendritic cell precursor from the CD11c+ cells expressing high levels of MHC class II molecules in the culture of bone marrow with FLT3 ligandHyunju In0Hyunju In1Ji Soo Park2Ji Soo Park3Hyun Soo Shin4Hyun Soo Shin5Seul Hye Ryu6Seul Hye Ryu7Seul Hye Ryu8Moah Sohn9Moah Sohn10Wanho Choi11Wanho Choi12Sejung Park13Sejung Park14Soomin Hwang15Soomin Hwang16Jeyun Park17Lihua Che18Lihua Che19Tae-Gyun Kim20Min Kyung Chu21Hye Young Na22Hye Young Na23Chae Gyu Park24Chae Gyu Park25Chae Gyu Park26Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaHeart-Immune-Brain Network Research Center, Department of Life Science, Ewha Womans University, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaBrain Korea 21 PLUS/FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Neurology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Neurology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of KoreaLaboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaHeart-Immune-Brain Network Research Center, Department of Life Science, Ewha Womans University, Seoul, Republic of KoreaLaboratory of Dendritic Cell Immunology, The Good Capital Institute for Immunology, Seoul, Republic of KoreaDendritic 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+MHCIIhi 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+MHCIIhi 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+MHCIIhi cells were within the classical DC2 (cDC2) gate. The analysis of FL-BM culture revealed that a majority of cDC2-gated CD11c+MHCIIhi cells exhibited a 2A1-CD83-CD115+CX3CR1+ phenotype, and the others consisted of 2A1+CD83+CD115-CX3CR1- and 2A1-CD83-CD115-CX3CR1- cells. According to the antigen uptake and presentation, morphologies, and gene expression profiles, 2A1-CD83-CD115-CX3CR1- cells were immature cDC2s and 2A1+CD83+CD115-CX3CR1- cells were mature cDC2s. Unexpectedly, however, 2A1-CD83-CD115+CX3CR1+ cells, the most abundant cDC2-gated MHCIIhi cell subset in FL-BM culture, were non-DCs. Adoptive cell transfer experiments in the FL-BM culture confirmed that the cDC2-gated MHCIIhi non-DCs were precursors to cDC2s, i.e., MHCIIhi pre-cDC2s. MHCIIhi pre-cDC2s also expressed the higher level of DC-specific transcription factor Zbtb46 as similarly as immature cDC2s. Besides, MHCIIhi 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 MHCIIhi pre-cDC2s are close lineage to cDCs. All in all, our study identified and characterized a new cDC precursor, exhibiting a CD11c+MHCIIhiCD115+CX3CR1+ phenotype, in FL-BM culture.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1179981/fullantigen presentationbone marrow cellscell differentiationcultured cellsdendritic cellsFLT3 ligand |
| spellingShingle | Hyunju In Hyunju In Ji Soo Park Ji Soo Park Hyun Soo Shin Hyun Soo Shin Seul Hye Ryu Seul Hye Ryu Seul Hye Ryu Moah Sohn Moah Sohn Wanho Choi Wanho Choi Sejung Park Sejung Park Soomin Hwang Soomin Hwang Jeyun Park Lihua Che Lihua Che Tae-Gyun Kim Min Kyung Chu Hye Young Na Hye Young Na Chae Gyu Park Chae Gyu Park Chae Gyu Park 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 Frontiers in Immunology antigen presentation bone marrow cells cell differentiation cultured cells dendritic cells FLT3 ligand |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | antigen presentation bone marrow cells cell differentiation cultured cells dendritic cells FLT3 ligand |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1179981/full |
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