A GeXP multiplex, RT-PCR assay was developed and optimized that simultaneously measures expression of a suite of immune-relevant genes in rainbow trout ( Oncorhynchus mykiss ), concentrating on tumor necrosis factor and interleukin-1 ligand/receptor systems and acute phase response genes. The dataset includes expression values for drpt , il11a , il1b1 , il1b2 , il1b3 , il1r-like-1 (e3-5), il1r-like-1 (e9-11), il1r1-like-a , il1r1-like-b , il1r2 , saa , tnfa1 , tnfa2 , tnfa3 , tnfrsf1a , tnfrsf1a-like-a , tnfrsf1a-like-b , tnfrsf5 , and tnfrsf9 . Gene expression was measured at four time-points post-challenge in both a resistant line (ARS-Fp-R) and a susceptible line (ARS-Fp-S) of rainbow trout. In addition, fish body weight, spleen index and the Flavobacterium psychrophilum load are reported. These data are an extension of information presented and discussed in " Proinflammatory cytokine and cytokine receptor gene expression kinetics following challenge with Flavobacterium psychrophilum in resistant and susceptible lines of rainbow trout ( Oncorhynchus mykiss )" (Kutyrev et al., 2016) [1].