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Christopher J Secombes - One of the best experts on this subject based on the ideXlab platform.
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Directional responses following Recombinant Cytokine stimulation of rainbow trout (Oncorhynchus mykiss) RTS-11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Christopher J SecombesAbstract:Background 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. 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. Results RNA 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. Conclusion We 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.
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directional responses following Recombinant Cytokine stimulation of rainbow trout oncorhynchus mykiss rts 11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Jun Zou, Christopher J SecombesAbstract: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. 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. RNA 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. We 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.
Samuel Am Martin - One of the best experts on this subject based on the ideXlab platform.
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Directional responses following Recombinant Cytokine stimulation of rainbow trout (Oncorhynchus mykiss) RTS-11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Christopher J SecombesAbstract:Background 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. 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. Results RNA 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. Conclusion We 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.
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directional responses following Recombinant Cytokine stimulation of rainbow trout oncorhynchus mykiss rts 11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Jun Zou, Christopher J SecombesAbstract: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. 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. RNA 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. We 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.
Daniela Frasca - One of the best experts on this subject based on the ideXlab platform.
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Regulation of Cytokine production in aging: Use of Recombinant Cytokines to upregulate mitogen-stimulated spleen cells
Mechanisms of Ageing and Development, 1997Co-Authors: Daniela Frasca, S Pucci, C Goso, P Barattini, S Barile, C Pioli, G DoriaAbstract:Abstract We investigated the production of IL-2 and IFN- γ (Th1 type) and IL-4 (Th2 type) Cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN- γ ), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN- γ and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN- γ , as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a Recombinant Cytokine (IL-1 β , IL-2, IL-3, IL-4, IL-12 or IFN- γ ) at various concentrations. It was found that Recombinant Cytokines by and large enhance Cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN- γ production as protein and mRNA. The addition of Recombinant Cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.
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Recombinant Cytokines as an approach to immune reconstitution in aging.
Developmental and comparative immunology, 1997Co-Authors: Daniela Frasca, Gino DohaAbstract:Abstract Aging is characterized by a decreased humoral and cell-mediated immunity to a large variety of exogenous antigens and by an increased propensity to autoantibody production, suggesting an age-related disregulation of the immune system. The decline in immune responsiveness to exogenous antigens has been attributed to thymus involution, consisting of a fall in the capacity to induce intrathymic T-cell growth and differentiation, and also to export mature T cells to the periphery. T-cell activation and secretion of soluble factors have been reported to change with aging, but, as with Cytokines, the results are conflicting. We investigated the production of IL-2 and IFN-γ (Th1 type) and IL-4 (Th2 type) Cytokines by mitogen-activated spleen cells from young, adult and old mice and their regulation by the addition of a Recombinant Cytokine (IL-1β, IL-2, IL-3, IL-4, IL-12, IFN-γ) at varying concentrations. The results indicate that Cytokine production can be enhanced only when it is deficient, suggesting the possible use of Recombinant Cytokines as efficient immunomodulators in age-associated immune disorders.
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Regulation of Cytokine production in aging: use of Recombinant Cytokines to upregulate mitogen-stimulated spleen cells.
Mechanisms of ageing and development, 1997Co-Authors: Daniela Frasca, S Pucci, C Goso, P Barattini, S Barile, C Pioli, G DoriaAbstract:We investigated the production of IL-2 and IFN-gamma (Th1 type) and IL-4 (Th2 type) Cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN-gamma), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN-gamma and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN-gamma, as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a Recombinant Cytokine (IL-1 beta, IL-2, IL-3, IL-4, IL-12 or IFN-gamma) at various concentrations. It was found that Recombinant Cytokines by and large enhance Cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN-gamma production as protein and mRNA. The addition of Recombinant Cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.
G Doria - One of the best experts on this subject based on the ideXlab platform.
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Regulation of Cytokine production in aging: Use of Recombinant Cytokines to upregulate mitogen-stimulated spleen cells
Mechanisms of Ageing and Development, 1997Co-Authors: Daniela Frasca, S Pucci, C Goso, P Barattini, S Barile, C Pioli, G DoriaAbstract:Abstract We investigated the production of IL-2 and IFN- γ (Th1 type) and IL-4 (Th2 type) Cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN- γ ), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN- γ and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN- γ , as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a Recombinant Cytokine (IL-1 β , IL-2, IL-3, IL-4, IL-12 or IFN- γ ) at various concentrations. It was found that Recombinant Cytokines by and large enhance Cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN- γ production as protein and mRNA. The addition of Recombinant Cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.
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Regulation of Cytokine production in aging: use of Recombinant Cytokines to upregulate mitogen-stimulated spleen cells.
Mechanisms of ageing and development, 1997Co-Authors: Daniela Frasca, S Pucci, C Goso, P Barattini, S Barile, C Pioli, G DoriaAbstract:We investigated the production of IL-2 and IFN-gamma (Th1 type) and IL-4 (Th2 type) Cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN-gamma), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN-gamma and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN-gamma, as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a Recombinant Cytokine (IL-1 beta, IL-2, IL-3, IL-4, IL-12 or IFN-gamma) at various concentrations. It was found that Recombinant Cytokines by and large enhance Cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN-gamma production as protein and mRNA. The addition of Recombinant Cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.
Dominic F Houlihan - One of the best experts on this subject based on the ideXlab platform.
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Directional responses following Recombinant Cytokine stimulation of rainbow trout (Oncorhynchus mykiss) RTS-11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Christopher J SecombesAbstract:Background 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. 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. Results RNA 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. Conclusion We 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.
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directional responses following Recombinant Cytokine stimulation of rainbow trout oncorhynchus mykiss rts 11 macrophage cells as revealed by transcriptome profiling
BMC Genomics, 2007Co-Authors: Samuel Am Martin, Dominic F Houlihan, Jun Zou, Christopher J SecombesAbstract: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. 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. RNA 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. We 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.