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Identification of genes responsive to solar simulated UV radiation in human monocyte-derived dendritic cells.

de la Fuente H, Lamana A, Mittelbrunn M, Perez-Gala S, Gonzalez S, García-Diez A, Vega M, Sanchez-Madrid F - PLoS ONE (2009)

Bottom Line: Ultraviolet (UV) irradiation has profound effects on the skin and the systemic immune system.Several genes involved in the regulation of the immune response were differentially regulated in UVA/UVB irradiated human monocyte-derived DCs, such as protein tyrosine phosphatase, receptor type E (PTPRE), thrombospondin-1 (THBS1), inducible costimulator ligand (ICOSL), galectins, Src-like adapter protein (SLA), IL-10 and CCR7.These results indicate that UV-exposure triggers the regulation of a complex gene repertoire involved in human-DC-mediated immune responses.

View Article: PubMed Central - PubMed

Affiliation: Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.

ABSTRACT
Ultraviolet (UV) irradiation has profound effects on the skin and the systemic immune system. Several effects of UV radiation on Dendritic cells (DCs) functions have been described. However, gene expression changes induced by UV radiation in DCs have not been addressed before. In this report, we irradiated human monocyte-derived DCs with solar-simulated UVA/UVB and analyzed regulated genes on human whole genome arrays. Results were validated by RT-PCR and further analyzed by Gene Set Enrichment Analysis (GSEA). Solar-simulated UV radiation up-regulated expression of genes involved in cellular stress and inflammation, and down-regulated genes involved in chemotaxis, vesicular transport and RNA processing. Twenty four genes were selected for comparison by RT-PCR with similarly treated human primary keratinocytes and human melanocytes. Several genes involved in the regulation of the immune response were differentially regulated in UVA/UVB irradiated human monocyte-derived DCs, such as protein tyrosine phosphatase, receptor type E (PTPRE), thrombospondin-1 (THBS1), inducible costimulator ligand (ICOSL), galectins, Src-like adapter protein (SLA), IL-10 and CCR7. These results indicate that UV-exposure triggers the regulation of a complex gene repertoire involved in human-DC-mediated immune responses.

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Western blot of selected proteins whose genes are differentially expressed in DCs.A, B. Human primary DCs were exposed to solar-simulated UV radiation as in Fig. 1, and cultured for a further 24 h before lysis and immunoblotting. Experiments were repeated at least three times. Specific protein bands were quantified by densitometry with respect to vimentin (loading control). A. Representative blots. B Results of densitometric analysis, presented as the ratio of expression in irradiated cells to that in non-irradiated cells. Data are the arithmetic means±SEM of three experiments. C. Human MCs and KCs were exposed to solar-simulated UV radiation and analyzed by western blot as in A. Data represents one of three experiments performed.
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pone-0006735-g003: Western blot of selected proteins whose genes are differentially expressed in DCs.A, B. Human primary DCs were exposed to solar-simulated UV radiation as in Fig. 1, and cultured for a further 24 h before lysis and immunoblotting. Experiments were repeated at least three times. Specific protein bands were quantified by densitometry with respect to vimentin (loading control). A. Representative blots. B Results of densitometric analysis, presented as the ratio of expression in irradiated cells to that in non-irradiated cells. Data are the arithmetic means±SEM of three experiments. C. Human MCs and KCs were exposed to solar-simulated UV radiation and analyzed by western blot as in A. Data represents one of three experiments performed.

Mentions: The functional outcome of altered gene expression results from changes in protein expression. However, protein expression cannot be reliably predicted from changes in mRNA levels. Moreover, the genes found to be differentially expressed have not been studied in detail in DCs. We therefore corroborated the RT-PCR results at protein level by western blot of DC lysates. A representative western blot of proteins encoded by genes regulated in DCs in response to UVA+UVB radiation is shown in Fig. 3A, and quantification of the changed expression relative to untreated cells is shown in Fig. 3B. Consistent with the RT-PCR data, solar-simulated UV-irradiation of human DCs resulted in clear down-regulation of galectin-1, galectin-3, SLA, PTPRE, ICOSL, and SOCS1. Again consistently, UV irradiation induced increases in the amounts of GADD45a and THBS1 proteins. As controls, expression of several proteins was determined in human melanocytes and keratinocytes (Fig. 3C). These blots confirm the absence of SLA protein from melanocytes, as well as the previously reported high expression of galectin-3 in these cells. The previously reported down-regulation of THBS1 in UV-irradiated human keratinocytes was also observed.


Identification of genes responsive to solar simulated UV radiation in human monocyte-derived dendritic cells.

de la Fuente H, Lamana A, Mittelbrunn M, Perez-Gala S, Gonzalez S, García-Diez A, Vega M, Sanchez-Madrid F - PLoS ONE (2009)

Western blot of selected proteins whose genes are differentially expressed in DCs.A, B. Human primary DCs were exposed to solar-simulated UV radiation as in Fig. 1, and cultured for a further 24 h before lysis and immunoblotting. Experiments were repeated at least three times. Specific protein bands were quantified by densitometry with respect to vimentin (loading control). A. Representative blots. B Results of densitometric analysis, presented as the ratio of expression in irradiated cells to that in non-irradiated cells. Data are the arithmetic means±SEM of three experiments. C. Human MCs and KCs were exposed to solar-simulated UV radiation and analyzed by western blot as in A. Data represents one of three experiments performed.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2727914&req=5

pone-0006735-g003: Western blot of selected proteins whose genes are differentially expressed in DCs.A, B. Human primary DCs were exposed to solar-simulated UV radiation as in Fig. 1, and cultured for a further 24 h before lysis and immunoblotting. Experiments were repeated at least three times. Specific protein bands were quantified by densitometry with respect to vimentin (loading control). A. Representative blots. B Results of densitometric analysis, presented as the ratio of expression in irradiated cells to that in non-irradiated cells. Data are the arithmetic means±SEM of three experiments. C. Human MCs and KCs were exposed to solar-simulated UV radiation and analyzed by western blot as in A. Data represents one of three experiments performed.
Mentions: The functional outcome of altered gene expression results from changes in protein expression. However, protein expression cannot be reliably predicted from changes in mRNA levels. Moreover, the genes found to be differentially expressed have not been studied in detail in DCs. We therefore corroborated the RT-PCR results at protein level by western blot of DC lysates. A representative western blot of proteins encoded by genes regulated in DCs in response to UVA+UVB radiation is shown in Fig. 3A, and quantification of the changed expression relative to untreated cells is shown in Fig. 3B. Consistent with the RT-PCR data, solar-simulated UV-irradiation of human DCs resulted in clear down-regulation of galectin-1, galectin-3, SLA, PTPRE, ICOSL, and SOCS1. Again consistently, UV irradiation induced increases in the amounts of GADD45a and THBS1 proteins. As controls, expression of several proteins was determined in human melanocytes and keratinocytes (Fig. 3C). These blots confirm the absence of SLA protein from melanocytes, as well as the previously reported high expression of galectin-3 in these cells. The previously reported down-regulation of THBS1 in UV-irradiated human keratinocytes was also observed.

Bottom Line: Ultraviolet (UV) irradiation has profound effects on the skin and the systemic immune system.Several genes involved in the regulation of the immune response were differentially regulated in UVA/UVB irradiated human monocyte-derived DCs, such as protein tyrosine phosphatase, receptor type E (PTPRE), thrombospondin-1 (THBS1), inducible costimulator ligand (ICOSL), galectins, Src-like adapter protein (SLA), IL-10 and CCR7.These results indicate that UV-exposure triggers the regulation of a complex gene repertoire involved in human-DC-mediated immune responses.

View Article: PubMed Central - PubMed

Affiliation: Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.

ABSTRACT
Ultraviolet (UV) irradiation has profound effects on the skin and the systemic immune system. Several effects of UV radiation on Dendritic cells (DCs) functions have been described. However, gene expression changes induced by UV radiation in DCs have not been addressed before. In this report, we irradiated human monocyte-derived DCs with solar-simulated UVA/UVB and analyzed regulated genes on human whole genome arrays. Results were validated by RT-PCR and further analyzed by Gene Set Enrichment Analysis (GSEA). Solar-simulated UV radiation up-regulated expression of genes involved in cellular stress and inflammation, and down-regulated genes involved in chemotaxis, vesicular transport and RNA processing. Twenty four genes were selected for comparison by RT-PCR with similarly treated human primary keratinocytes and human melanocytes. Several genes involved in the regulation of the immune response were differentially regulated in UVA/UVB irradiated human monocyte-derived DCs, such as protein tyrosine phosphatase, receptor type E (PTPRE), thrombospondin-1 (THBS1), inducible costimulator ligand (ICOSL), galectins, Src-like adapter protein (SLA), IL-10 and CCR7. These results indicate that UV-exposure triggers the regulation of a complex gene repertoire involved in human-DC-mediated immune responses.

Show MeSH
Related in: MedlinePlus