Abstract
BackgroundThe early stages of the immune response are regulated by key cytokines including both interleukin 1β (IL-1β) and interferon-γ (IFN-γ) which stimulate panels of responsive genes via conserved signal transduction pathways. To further our understanding of the transcriptional response to these cytokines in lower vertebrates we have utilized microarray analysis to characterize the transcriptional response to recombinant rainbow trout IL-1β and IFN-γ in the trout macrophage cell line RTS-11.ResultsRNA was extracted from stimulated or control cells following 6 h incubation and used to hybridize to a salmonid cDNA microarray containing 16,006 different genes. Analysis of the arrays revealed mRNA transcripts that were differentially expressed as a result of exposure to the recombinant proteins, with some responses common for both cytokines. In general the recombinant IL-1β elicited a response where genes involved in the acute phase response were up-regulated, whilst the recombinant IFN-γ induced strong up-regulation of genes involved in the MHC class I antigen presentation pathway. Key genes were chosen that were differentially regulated and analysed by real time PCR at additional time points, up to 48 h following stimulation. This allowed a deeper insight into the kinetics of the response to the cytokines in this cell line.ConclusionWe demonstrated that in fish both rIL-1β and rIFN-γ stimulated discrete panels of mRNA transcripts which indicted the cells were being directed towards different cellular functions, with IL-β inducing genes involved in the inflammatory response, whereas IFN-γ induced genes associated with antigen presentation.
Highlights
The early stages of the immune response are regulated by key cytokines including both interleukin 1β (IL-1β) and interferon-γ (IFN-γ) which stimulate panels of responsive genes via conserved signal transduction pathways
BMC Genomics 2007, 8:150 http://www.biomedcentral.com/1471-2164/8/150 large number are known from a wide variety of species including trout, salmon, carp and zebrafish amongst others, and including many interleukins e.g. IL-1 [6,7], IL-2 [8], IL-6 [9], IL-8 [10], IL-10 [11], IL-11 [12], type 1 and type 2 interferons [13,14,15], lymphotoxin β [16], transforming growth factors [17], chemokines [18,19], as well as some with no obvious homology to known mammalian cytokines
Cell stimulation To dissect the immunomodulatory effects of the rainbow trout recombinant proteins on RTS-11 cells, the cells were treated with either 20 ng ml-1 rIL-1β or rIFN-γ protein
Summary
The early stages of the immune response are regulated by key cytokines including both interleukin 1β (IL-1β) and interferon-γ (IFN-γ) which stimulate panels of responsive genes via conserved signal transduction pathways. BMC Genomics 2007, 8:150 http://www.biomedcentral.com/1471-2164/8/150 large number are known from a wide variety of species including trout, salmon, carp and zebrafish amongst others, and including many interleukins e.g. IL-1 [6,7], IL-2 [8], IL-6 [9], IL-8 [10], IL-10 [11], IL-11 [12], type 1 and type 2 interferons [13,14,15], lymphotoxin β [16], transforming growth factors [17], chemokines [18,19], as well as some with no obvious homology to known mammalian cytokines. The microarrays used in the above studies use both cDNA and oligo array type platforms
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