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Both host and parasite MIF molecules bind to chicken macrophages via CD74 surface receptor

Author:
Kim, Sungwon, Cox, Chasity M., Jenkins, Mark C., Fetterer, Ray H., Miska, Katarzyna B., Dalloul, Rami A.
Source:
Developmental and comparative immunology 2014 v.47 no.2 pp. 319-326
ISSN:
0145-305X
Subject:
Eimeria, adaptive immunity, chickens, fibroblasts, flow cytometry, fluorescence, fluorescent antibody technique, green fluorescent protein, inflammation, innate immunity, macrophage migration inhibitory factors, macrophages, monocytes, parasites, recombinant proteins, staining, transmembrane proteins
Abstract:
Macrophage migration inhibitory factor (MIF) is recognized as a soluble protein that inhibits the random migration of macrophages and plays a pivotal immunoregulatory function in innate and adaptive immunity. Our group has identified both chicken and Eimeria MIFs, and characterized their function in enhancing innate immune responses during inflammation. In this study, we report that chicken CD74 (ChCD74), a type II transmembrane protein, functions as a macrophage surface receptor that binds to MIF molecules. First, to examine the binding of MIF to chicken monocytes/macrophages, fresh isolated chicken peripheral blood mononuclear cells (PBMCs) were stimulated with rChIFN-γ and then incubated with recombinant chicken MIF (rChMIF). Immunofluorescence staining with anti-ChMIF followed by flow cytometry revealed the binding of MIF to stimulated PBMCs. To verify that ChCD74 acts as a surface receptor for MIF molecules, full-length ChCD74p41 was cloned, expressed and its recombinant protein (rChCD74p41) transiently over-expressed with green fluorescent protein in chicken fibroblast DF-1 cells. Fluorescence analysis revealed a higher population of cells double positive for CD74p41 and rChMIF, indicating the binding of rChMIF to DF-1 cells via rChCD74p41. Using a similar approach, it was found that Eimeria MIF (EMIF), which is secreted by Eimeria sp. during infection, bound to chicken macrophages via ChCD74p41 as a surface receptor. Together, this study provides conclusive evidence that both host and parasite MIF molecules bind to chicken macrophages via the surface receptor ChCD74.
Agid:
5335461