title: Development of a capture ELISA to determine kinetics of soluble CD25 following in vitro and in vivo stimulation of duck peripheral blood monocytes.
abstract: In humans and other mammals, the α-chain of interleukin-2 (IL-2) receptor (CD25) is induced and expressed on the cell surface after lymphocyte activation and is released from the membrane of activated cells as a smaller soluble form (sCD25). However, little is known about avian sCD25. In the present study, we developed an antigen capture enzyme-linked immunosorbent assay (AC-ELISA) to detect serum sCD25 in ducks, and we used flow cytometry (FCM) to analyze the frequency of CD25(+) cells in the peripheral blood of ducks infected with H9N2 or H5N1 avian influenza virus (AIV) or serotype II Riemerella anatipestifer (RA). Using the AC-ELISA, duck sCD25 molecules were detected in the supernatant and lysates of concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC), and in the serum of ducks infected with H5N1 virus and RA. However, no sCD25 was detected in the serum of H9N2 AIV-infected ducks. FCM analysis revealed that CD25(+) cells were upregulated within the PBMC of RA-infected ducks throughout the experiment until death, while in the PBMC of H9N2- and H5N1 AIV-infected ducks, the frequency of CD25(+) cells increased in the early stage of infection and then returned to a lower level. Our findings confirm that the dynamics of sCD25 and CD25(+) cells are different in the peripheral blood of ducks infected with H9N2 virus, H5N1 virus, and RA.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, PR China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Fang
authors.forename: Jianyou
authors.initials: J
authors.lastname: Gu
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D003208:Concanavalin A / Q000276:immunology; D004372:Ducks / Q000097:blood / Q000276:immunology* / Q000382:microbiology / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D041965:Flavobacteriaceae / Q000276:immunology; D045826:Flavobacteriaceae Infections / Q000276:immunology / Q000662:veterinary; D005434:Flow Cytometry / Q000662:veterinary; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology; D053127:Influenza A Virus, H9N2 Subtype / Q000276:immunology; D005585:Influenza in Birds / Q000276:immunology; D053645:Interleukin-2 Receptor alpha Subunit / Q000096:biosynthesis* / Q000097:blood*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D018655:Lymphocyte Count / Q000662:veterinary; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: D053645:Interleukin-2 Receptor alpha Subunit; D003208:Concanavalin A