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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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Gene expression induced by UV irradiation in human primary DCs, MCs and KCs.Human primary monocyte-derived DCs (moDCs), melanocytes (MCs) and keratinocytes (KCs) were exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB) as in Fig. 1 and total RNA was extracted after a further 6 h in culture. Primer sequences are shown in Table S1. Expression levels were normalized to 18s RNA. A. Genes encoding cytokines and chemokine receptors. B. Genes related to DNA damage and p53 response. C. Genes potentially involved in immunomodulation. Results are shown as log10 of fold up-regulation or down-regulation in UV-irradiated cells compared with expression in non-irradiated controls. Data correspond to arithmetic mean±SEM.
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pone-0006735-g002: Gene expression induced by UV irradiation in human primary DCs, MCs and KCs.Human primary monocyte-derived DCs (moDCs), melanocytes (MCs) and keratinocytes (KCs) were exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB) as in Fig. 1 and total RNA was extracted after a further 6 h in culture. Primer sequences are shown in Table S1. Expression levels were normalized to 18s RNA. A. Genes encoding cytokines and chemokine receptors. B. Genes related to DNA damage and p53 response. C. Genes potentially involved in immunomodulation. Results are shown as log10 of fold up-regulation or down-regulation in UV-irradiated cells compared with expression in non-irradiated controls. Data correspond to arithmetic mean±SEM.

Mentions: To identify genes regulated by UV irradiation specifically in DCs, we added a further 11 genes to the set for testing by TaqMan-based RT-PCR: IL-10, GADD45A, GADD45B, Galectin 1, Galectin 3, SLA, CXCR4, SOCS1, PTPRE, CCR7, and IL12A (Table S1). Although the modulation of these genes was not detected by the microarrays experiments, some of them were highlighted by GSEA, others genes like PTPRE, SLA and GADD45A and B were modulated by UV irradiation although with a p value>0.05. All of these genes were chosen mainly because of their potential immunomodulatory roles. The regulation by UV radiation of the complete set of 24 genes was studied in DCs obtained from 6 new donors and compared with human primary keratinocytes and melanocytes. The irradiation dose was the same for the 3 human cells types; cell apoptosis was not detected at time of RT-PCR analysis (data not shown). The 24 genes were classified into three functional groups: i) genes encoding cytokines and chemokine receptors (Fig. 2A); ii) genes related to DNA damage and p53 responses (Fig. 2B); and iii) genes potentially involved in immunomodulation (Fig. 2C). We included the p53 target gene GADD45a with the immunoregulatory genes because there is increasing evidence that it has important functions in the immune system in addition to its roles in cell-cycle arrest, DNA repair, and cell survival [17].


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)

Gene expression induced by UV irradiation in human primary DCs, MCs and KCs.Human primary monocyte-derived DCs (moDCs), melanocytes (MCs) and keratinocytes (KCs) were exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB) as in Fig. 1 and total RNA was extracted after a further 6 h in culture. Primer sequences are shown in Table S1. Expression levels were normalized to 18s RNA. A. Genes encoding cytokines and chemokine receptors. B. Genes related to DNA damage and p53 response. C. Genes potentially involved in immunomodulation. Results are shown as log10 of fold up-regulation or down-regulation in UV-irradiated cells compared with expression in non-irradiated controls. Data correspond to arithmetic mean±SEM.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2727914&req=5

pone-0006735-g002: Gene expression induced by UV irradiation in human primary DCs, MCs and KCs.Human primary monocyte-derived DCs (moDCs), melanocytes (MCs) and keratinocytes (KCs) were exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB) as in Fig. 1 and total RNA was extracted after a further 6 h in culture. Primer sequences are shown in Table S1. Expression levels were normalized to 18s RNA. A. Genes encoding cytokines and chemokine receptors. B. Genes related to DNA damage and p53 response. C. Genes potentially involved in immunomodulation. Results are shown as log10 of fold up-regulation or down-regulation in UV-irradiated cells compared with expression in non-irradiated controls. Data correspond to arithmetic mean±SEM.
Mentions: To identify genes regulated by UV irradiation specifically in DCs, we added a further 11 genes to the set for testing by TaqMan-based RT-PCR: IL-10, GADD45A, GADD45B, Galectin 1, Galectin 3, SLA, CXCR4, SOCS1, PTPRE, CCR7, and IL12A (Table S1). Although the modulation of these genes was not detected by the microarrays experiments, some of them were highlighted by GSEA, others genes like PTPRE, SLA and GADD45A and B were modulated by UV irradiation although with a p value>0.05. All of these genes were chosen mainly because of their potential immunomodulatory roles. The regulation by UV radiation of the complete set of 24 genes was studied in DCs obtained from 6 new donors and compared with human primary keratinocytes and melanocytes. The irradiation dose was the same for the 3 human cells types; cell apoptosis was not detected at time of RT-PCR analysis (data not shown). The 24 genes were classified into three functional groups: i) genes encoding cytokines and chemokine receptors (Fig. 2A); ii) genes related to DNA damage and p53 responses (Fig. 2B); and iii) genes potentially involved in immunomodulation (Fig. 2C). We included the p53 target gene GADD45a with the immunoregulatory genes because there is increasing evidence that it has important functions in the immune system in addition to its roles in cell-cycle arrest, DNA repair, and cell survival [17].

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