Chemokines not only act as chemoattractants for immune cells, but also exert immunomodulatory actions, thus modulating the immune functions of their target cells. In rainbow trout (Oncorhynchus mykiss), twenty-four chemokines have been identified to date. Even though their sequences have been reported, their biological role has been scarcely elucidated and the role that these chemokines have on the antiviral response in fish has been poorly studied. In this sense, in the current work, we have determined the levels of expression of several of these rainbow trout chemokines (CXCd, gammaIP, CK1, CK3, CK5B, CK6, CK7A, CK9 and CK12) in head kidney and spleen during the course of a viral infection using two different viruses, viral hemorrhagic septicaemia virus (VHSV) and infectious pancreatic necrosis virus (IPNV), comparing them to the levels induced by poly I:C. We also determined the effects that the two viruses and poly I:C provoked on the levels of expression of these chemokines in vitro in head kidney leucocytes. Overall, VHSV was capable of modulating gammaIP, CXCd, CK1, CK3, CK5B, CK6 and CK12, while IPNV induced a very different chemokine profile and affected CK1, CK5B, CK6, CK7A, CK9 and CK12. On the other hand, a viral mimic such as poly I:C was capable of up-regulating gammaIP, CXCd, CK1, CK3, CK5B, CK7A and CK12. As more information becomes available concerning the immune role and target cells that these chemokines have on rainbow trout, we would be able to better interpret the importance of these differences in the pathogenicity of these two viruses.