Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterization of avian conventional dendritic cells. Issue 2 (30th November 2021)

• Record Type: Journal Article
• Authors: Wu, Zhiguang; Hu, Tuanjun; Chintoan‐Uta, Cosmin; Macdonald, Joni; Stevens, Mark P.; Sang, Helen; Hume, David A.; Kaiser, Pete; Balic, Adam
• Is Part Of: Immunology. Volume 165:Issue 2(2022)
• Page Start: 171
• Page End: 194
• Publication Date: 2021-11-30
• Subjects: avian -- chicken -- conventional dendritic cell -- CSF1R -- FLT3 -- macrophage -- reagents -- XCR1; Immunology -- Periodicals
• DOI: 10.1111/imm.13426
• Languages: English
• ISSNs: 0019-2805

Abstract

Abstract: Conventional dendritic cells (cDC) are bone marrow‐derived immune cells that play a central role in linking innate and adaptive immunity. cDCs efficiently uptake, process and present antigen to naïve T cells, driving clonal expansion of antigen‐specific T‐cell responses. 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 FLT3 cDNA was cloned and a monoclonal antibody to cell surface FLT3 was generated. This antibody identified a distinct FLT3 HI splenic subset which lack expression of signature markers for B cells, T cells or monocyte/macrophages. By combining anti‐FLT3 and CSF1R ‐eGFP transgenic expression, three major populations within the mononuclear phagocyte system were identified in the spleen. The cDC1 subset 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 AF647 ). Flow cytometry staining of XCL1 AF647 on splenocytes showed that all chicken FLT3 HI cells exclusively express XCR1, supporting the hypothesis that this population comprises bona fide 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 + 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. Abstract : We have developed tools to specifically identify chicken conventional dendritic cells (cDCs). Chicken cDCs are express high levels of FLT3 and XCR1. While the resemble the mammalian cDC1 subset, significant differences exist.