Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterization of avian conventional dendritic cells.
Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterization of avian conventional dendritic cells.
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DOI：
10.1111/imm.13426
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发表时间：
2022-02
期刊：
影响因子：
6.4
通讯作者：
Balic, Adam
中科院分区：
文献类型：
--
作者：
Wu, Zhiguang;Hu, Tuanjun;Chintoan-Uta, Cosmin;Macdonald, Joni;Stevens, Mark P.;Sang, Helen;Hume, David A.;Kaiser, Pete;Balic, Adam
传统的树突状细胞（cDC）是骨髓来源的免疫细胞，在连接先天性免疫和适应性免疫中发挥核心作用。cDC有效地摄取、加工并将抗原呈递给幼稚T细胞，从而驱动抗原特异性T细胞应答的克隆扩增。在鸡中，缺乏有效和精确鉴定cDC的重要试剂。在这项研究中，我们已经开发了几种新的试剂用于鉴定和表征鸡cDCs。克隆鸡FLT 3 cDNA，并产生针对细胞表面FLT 3的单克隆抗体。该抗体鉴定了一个独特的FLT 3 HI脾脏亚群，该亚群缺乏B细胞、T细胞或单核细胞/巨噬细胞的标志物的表达。通过结合抗FLT 3和CSF 1 R-eGFP转基因表达，在脾脏中鉴定了单核吞噬细胞系统内的三个主要群体。哺乳动物cDC的cDC 1亚群表达趋化因子受体XCR 1。为了表征鸡cDC，开发了与Alexa Fluor 647缀合的合成鸡趋化因子（C基序）配体（XCL 1）肽（XCL 1AF 647）。脾细胞上XCL 1AF 647的流式细胞术染色显示所有鸡FLT 3 HI细胞仅表达XCR 1，支持该群体包含真正的鸡cDC的假设。进一步分析显示，鸡cDC表达CSF 1 R，但缺乏CSF 2 R的表达。总的来说，鸡cDC的细胞表面表型与哺乳动物XCR 1 + cDC 1部分保守，与哺乳动物直系同源物相比，CSF 1 R和CSF 2 R表达存在明显差异。这些原始试剂允许有效地鉴定鸡cDC，以研究其在鸡免疫和疾病中的重要作用。我们已经开发了专门识别鸡常规树突状细胞（cDC）的工具。鸡cDC表达高水平的FLT 3和XCR 1。虽然类似于哺乳动物cDC 1亚群，但存在显著差异。
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 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 characterize 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 with mammalian orthologues. These original reagents allow the efficient identification of chicken cDCs to investigate their important roles in the chicken immunity and diseases. 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.
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DOI：
10.1084/jem.20030107
发表时间：
2003-07-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
D'Amico A;Wu L
通讯作者：
Wu L
DOI：
10.1084/jem.163.4.872
发表时间：
1986-04-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Bowers WE;Berkowitz MR
通讯作者：
Berkowitz MR
DOI：
10.1084/jem.192.12.1685
发表时间：
2000-12-18
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
den Haan JM;Lehar SM;Bevan MJ
通讯作者：
Bevan MJ
影响因子：
30.5
作者：
Bedoui, Sammy;Whitney, Paul G.;Heath, William R.
通讯作者：
Heath, William R.
影响因子：
4.4
作者：
Cheminay, C;Möhlenbrink, A;Hensel, M
通讯作者：
Hensel, M