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In chicken, vital reagents are lacking for the efficient and precise identification of cDCs. In this study, we have developed several novel reagents for the identification and characterization of chicken cDCs. Chicken <jats:italic>FLT3<\/jats:italic> cDNA was cloned and a monoclonal antibody to cell surface FLT3 was generated. This antibody identified a distinct FLT3<jats:sup>HI<\/jats:sup> splenic subset which lack expression of signature markers for B cells, T cells or monocyte\/macrophages. By combining anti\u2010FLT3 and <jats:italic>CSF1R<\/jats:italic>\u2010eGFP transgenic expression, three major populations within the mononuclear phagocyte system were identified in the spleen. The cDC1\u00a0subset of mammalian cDCs express the chemokine receptor XCR1. To characterize chicken cDCs, a synthetic chicken chemokine (C motif) ligand (XCL1) peptide conjugated to Alexa Fluor 647 was developed (XCL1<jats:sup>AF647<\/jats:sup>). Flow cytometry staining of XCL1<jats:sup>AF647<\/jats:sup> on splenocytes showed that all chicken FLT3<jats:sup>HI<\/jats:sup> cells exclusively express XCR1, supporting the hypothesis that this population comprises <jats:italic>bona fide<\/jats:italic> chicken cDCs. Further analysis revealed that chicken cDCs expressed CSF1R but lacked the expression of CSF2R. Collectively, the cell surface phenotypes of chicken cDCs were partially conserved with mammalian XCR1<jats:sup>+<\/jats:sup> cDC1, with distinct differences in CSF1R and CSF2R expression compared with mammalian orthologues. These original reagents allow the efficient identification of chicken cDCs to investigate their important roles in the chicken immunity and diseases.<\/jats:p>","DOI":"10.1111\/imm.13426","type":"journal-article","created":{"date-parts":[[2021,11,12]],"date-time":"2021-11-12T17:33:20Z","timestamp":1636738400000},"page":"171-194","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterization of avian conventional dendritic cells"],"prefix":"10.1111","volume":"165","author":[{"given":"Zhiguang","family":"Wu","sequence":"first","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuanjun","family":"Hu","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cosmin","family":"Chintoan\u2010Uta","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joni","family":"Macdonald","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark P.","family":"Stevens","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen","family":"Sang","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Hume","sequence":"additional","affiliation":[{"name":"Translational Research Institute Mater Research Institute\u2010University of Queensland  Woolloongabba Qld Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pete","family":"Kaiser","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8369-7269","authenticated-orcid":false,"given":"Adam","family":"Balic","sequence":"additional","affiliation":[{"name":"The Roslin Institute University of Edinburgh, Easter Bush  Midlothian UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,11,30]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00456-1"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-020711-074950"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.04.016"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-020-00413-x"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2017.8"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.aaw1941"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2008.08.006"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1080\/03079451003786105"},{"key":"e_1_2_11_10_1","doi-asserted-by":"crossref","first-page":"2626","DOI":"10.4049\/jimmunol.125.6.2626","article-title":"Gut\u2010associated lymphoid tissue in the chicken. 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