CpG-oligodeoxynucleotide-stimulated chicken heterophil degranulation is serum cofactor and cell surface receptor dependent

Synthetic oligodeoxynucleotide containing unmethylated CpG motif (CpG-ODN) is immune stimulatory to chicken heterophils. Recognition of CpG-ODN by chicken heterophils leads to the mobilization and release of granules. This CpG-ODN-induced heterophil degranulation was chicken serum (CS)-dependent. Heat-denaturation and membrane filtration of CS revealed that the active serum cofactor(s) was likely a protein in nature with a molecule mass within 50,000 to 100,000. This serum cofactor(s) was heat-resistant at 56 °C for 1 h. The involvement of a cell surface receptor in recognition of CpG-ODN was also demonstrated by (1) trypsin treatment of the heterophils abrogated the degranulation response and (2) CpG-ODN-induced heterophil degranulation was sensitive to the inhibition of Clathrin-dependent endocytosis. In addition, among various microbial agonists, including CpG-ODN, lipopolysaccharide, lipoteichoic acid, phorbol myristate acetate, and formalin-killed Salmonella enteritidis, CpG-ODN was the only agonist that displayed serum-dependent induction of degranulation in chicken heterophils. This is the first report that shows serum-dependent activation of leukocytes by CpG-ODN.