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Isolation and Characterization of Interleukin‐1 From Bovine Polymorphonuclear Leukocytes
Author(s) -
Canning Peter C.,
Neill John D.
Publication year - 1989
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.45.1.21
Subject(s) - zymosan , biology , phytohaemagglutinin , isoelectric focusing , size exclusion chromatography , granulocyte , opsonin , microbiology and biotechnology , cell culture , isoelectric point , interleukin , interleukin 2 , immunology , biochemistry , immune system , cytokine , phagocytosis , in vitro , enzyme , genetics
Interleukin‐1 (IL‐1α and IL‐1β collectively) has been shown to be produced by a wide variety of cell types. The purpose of this study was to evaluate the ability of bovine polymorphonuclear leukocytes (PMNs) to synthesize and release IL‐1‐like cytokines and characterize the active molecule(s). Purified peripheral blood PMNs were cultured for various periods of time in the presence of opsonized zymosan particles. The resulting culture supernatants exhibited IL‐1 activity as determined by enhanced mitogen‐induced proliferation of the D10 G4.1 murine T‐helper cell line. Supernatants from nonstimulated PMNs or PMNs stimulated for less than 6 h did not enhance D10 G4.1 proliferation. The active molecule (PMNIL‐1) was isolated by using gel filtration high‐performance liquid chromatography (HPLC). Further characterization of the HPLC‐purtfied molecule by SDS‐PAGE and isoelectric focusing indicates bovine PMNIL‐1 has a molecular weight of 17.6 kd and a pl of 4.1.