The biofunction of orange-spotted grouper (Epinephelus coioides) CC chemokine ligand 4 (CCL4) in innate and adaptive immunity

CC chemokine (motif) ligand 4 (CCL4) is indispensable to the chemoattraction of macrophages, natural killer cells, and lymphocytes in mammals; however, it has only been cloned in a limited number of fish species and information related to its biofunction remains ambiguous with regard to teleosts. To explore the role of teleost CCL4, we first evaluated the mRNA expression of the Epinepheltts coioides CCL4 (gCCL4) gene in various organs under LPS and poly (I:C) stimulated; secondary, we evaluated the immune-related genes expression of fish under the recombinant gCCL4 protein stimulated. Our results revealed an increase in the mRNA of gCCL4 in immune organs immediately following stimulation by poly (I:C); however, in LPS stimulated fish, the expression did not increase until nearly 24 h after induction. In biofunction assays, recombinant gCCL4 was found to induce chemotactic activity in the peripheral blood leukocytes of groupers and up-regulate the gene expressions of grouper TNFA1 (TNF-alpha 1), TNFA2 (TNF-alpha 2), IFNG (IFN-gamma), MX, TBX21 (1-bet), CD8 (alpha and beta chain). These findings indicate that grouper CCL4 attracts leukocytes, induces an inflammatory response, and drives lymphocyte differentiation into the Th1 pathway. (C) 2013 Elsevier Ltd. All rights reserved.