Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterisation of avian conventional dendritic cells

Zhiguang Wu, Tuanjun Hu, Cosmin Chintoan-Uta, Joni Macdonald, Mark P Stevens, Helen Sang, David A Hume, Pete Kaiser, Adam Balic

Research output: Contribution to journalArticlepeer-review

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 characterisation of chicken cDCs. Chicken FLT3 cDNA was cloned and a monoclonal antibody to cell surface FLT3 was generated. This antibody identified a distinct FLT3HI 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 characterise chicken cDCs, a synthetic chicken chemokine (C motif) ligand (XCL1) peptide conjugated to Alexa Fluor 647 was developed (XCL1AF647 ). Flow cytometry staining of XCL1AF647 on splenocytes showed that all chicken FLT3HI 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 to mammalian orthologues. These original reagents allow the efficient identification of chicken cDCs to investigate their important roles in the chicken immunity and diseases.

Original languageEnglish
Pages (from-to)171-194
JournalImmunology
Volume165
Early online date12 Nov 2021
DOIs
Publication statusE-pub ahead of print - 12 Nov 2021

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