Differential DARC/ACKR1 expression distinguishes venular from non-venular endothelial cells in murine tissues

BMC Biology

Methodology Article

Antal Rot¹  Igor Novitzky-Basso²  Carolina Perdomo³  Olga Barreiro³  Irina Mazo³  Aude Thiriot³  Ulrich H. von Andrian³  Jamie K. Kishimoto³  Guiying Cheng³  Klaus Ley⁴  Sara McArdle⁴  Robinson Triboulet⁵ 

[1] Center for Immunology and Infection, Department of Biology, University of York, YO10 5DD, Heslington, York, UK;Center for Immunology and Infection, Department of Biology, University of York, YO10 5DD, Heslington, York, UK;Present address: Blood and Marrow Transplant Unit, Queen Elizabeth University Hospital, Glasgow, UK;Department of Microbiology and Immunobiology & HMS Center for Immune Imaging, Harvard Medical School, 77 Avenue Louis Pasteur, 02115, Boston, MA, USA;The Ragon Institute of MGH, MIT and Harvard, 02139, Cambridge, MA, USA;Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA;Stem Cell Program, Boston Children’s Hospital, Boston, MA, USA;

关键词: Monoclonal antibody;    Microvascular endothelium;    Leukocyte adhesion;    DARC/ACKR1;    Chemokines;   

DOI  :  10.1186/s12915-017-0381-7

 received in 2017-01-10, accepted in 2017-04-26,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundIntravascular leukocyte recruitment in most vertebrate tissues is restricted to postcapillary and collecting venules, whereas capillaries and arterioles usually support little or no leukocyte adhesion. This segmental restriction is thought to be mediated by endothelial, rather than hemodynamic, differences. The underlying mechanisms are largely unknown, in part because effective tools to distinguish, isolate, and analyze venular endothelial cells (V-ECs) and non-venular endothelial cells (NV-ECs) have been unavailable. We hypothesized that the atypical chemokine receptor DARC (Duffy Antigen Receptor for Chemokines, a.k.a. ACKR1 or CD234) may distinguish V-ECs versus NV-ECs in mice.MethodsWe generated a rat-anti-mouse monoclonal antibody (MAb) that specifically recognizes the erythroid and endothelial forms of native, surface-expressed DARC. Using this reagent, we characterized DARC expression and distribution in the microvasculature of murine tissues. ResultsDARC was exquisitely restricted to post-capillary and small collecting venules and completely absent from arteries, arterioles, capillaries, veins, and most lymphatics in every tissue analyzed. Accordingly, intravital microscopy showed that adhesive leukocyte-endothelial interactions were restricted to DARC+ venules. DARC was detectable over the entire circumference of V-ECs, but was more concentrated at cell-cell junctions. Analysis of single-cell suspensions suggested that the frequency of V-ECs among the total microvascular EC pool varies considerably between different tissues.ConclusionsImmunostaining of endothelial DARC allows the identification and isolation of intact V-ECs from multiple murine tissues. This strategy may be useful to dissect the mechanisms underlying segmental microvascular specialization in healthy and diseased tissues and to characterize the role of EC subsets in tissue-homeostasis, immune surveillance, infection, inflammation, and malignancies.

【 授权许可】

CC BY   
© von Andrian et al. 2017