Chicken meat and eggs contaminated with Salmonella result in economic losses in the poultry industry and potential human infection. Intestinal parasites have been shown to lead to a reduction in the utilization of nutrients and performance in poultry. This study provides insight into the immune responses used by hens of 3 genetically distinct chicken lines (broiler, Leghorn, and Fayoumi) in the presence and absence of Salmonella Enteritidis infection. Understanding the range of immune responses used by different lines in response to Salmonella Enteritidis may help the poultry industry genetically select birds that are more pathogen resistant. The splenic mRNA levels of several immune-related genes [IL-6, IL-8, IL-10, IL-18, macrophage inflammatory protein 1β, interferon (IFN)-γ, transforming growth factor β1, and regulated upon activation, normal T cell expressed, and secreted (RANTES)] were analyzed by quantitative PCR. Line, challenge, and their interaction were considered fixed effects. Line had a significant effect on the mRNA expression of RANTES (P < 0.02) and IFN-γ (P < 0.03). Broilers expressed significantly more splenic RANTES mRNA than Fayoumis, and significantly more splenic IFN-γ mRNA than Leghorns (P < 0.05). There was a significant interaction of genetic line and challenge on IL-18 (P < 0.02) and IL-6 (P < 0.01) mRNA expression. Although there was a significant interaction of genetic line and challenge for IL-18, Tukey's test analysis only showed differences at a suggestive level (P < 0.1). Bacterial challenge had a significant effect on IL-6 mRNA expression only within the Fayoumi line. Challenged Fayoumis expressed significantly less IL-6 mRNA than nonchallenged Fayoumis (P < 0.05). The observed differences in mRNA expression of selected cytokines support the concept that these distinct genetic lines utilize different immune responses at homeostasis and in response to Salmonella Enteritidis infection.