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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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Analysis of gene expression in UV-irradiated human DCs by RT-PCR.Real-time semiquantitative TaqMan RT-PCR was performed to validate differential gene expression induced by UV irradiation in human monocyte-derived dendritic cells (DCs) in culture. DCs (obtained from 3 donors) were untreated or exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB). 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. Bars correspond to log10 of fold down-regulation or up-regulation compared with non-irradiated cells.
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pone-0006735-g001: Analysis of gene expression in UV-irradiated human DCs by RT-PCR.Real-time semiquantitative TaqMan RT-PCR was performed to validate differential gene expression induced by UV irradiation in human monocyte-derived dendritic cells (DCs) in culture. DCs (obtained from 3 donors) were untreated or exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB). 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. Bars correspond to log10 of fold down-regulation or up-regulation compared with non-irradiated cells.

Mentions: To corroborate the microarray results, 15 of the modulated genes were selected for analysis by TaqMan-based RT-PCR. Table S1 lists the primers and probes for the genes selected for this analysis. Among these genes are several reported to play roles in the regulation of the immune response, including CD163, inducible costimulator ligand (ICOSL), IL1, Src-like adapter protein (SLA) and thrombospondin-1 (THBS1). As controls, we included several stress-response genes known to be modulated in response to UV irradiation (GDF15, FDXR, PLK2, PCNA). The RT-PCR analysis was carried out with the same RNA samples used for the microarrays. The results confirm the altered expression of all the genes tested except for AICDA and CD163 (Fig. 1).


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)

Analysis of gene expression in UV-irradiated human DCs by RT-PCR.Real-time semiquantitative TaqMan RT-PCR was performed to validate differential gene expression induced by UV irradiation in human monocyte-derived dendritic cells (DCs) in culture. DCs (obtained from 3 donors) were untreated or exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB). 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. Bars correspond to log10 of fold down-regulation or up-regulation compared with non-irradiated cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2727914&req=5

pone-0006735-g001: Analysis of gene expression in UV-irradiated human DCs by RT-PCR.Real-time semiquantitative TaqMan RT-PCR was performed to validate differential gene expression induced by UV irradiation in human monocyte-derived dendritic cells (DCs) in culture. DCs (obtained from 3 donors) were untreated or exposed to solar-simulated UV radiation (3.7 J/cm2 UVA+0.3 J/cm2 UVB). 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. Bars correspond to log10 of fold down-regulation or up-regulation compared with non-irradiated cells.
Mentions: To corroborate the microarray results, 15 of the modulated genes were selected for analysis by TaqMan-based RT-PCR. Table S1 lists the primers and probes for the genes selected for this analysis. Among these genes are several reported to play roles in the regulation of the immune response, including CD163, inducible costimulator ligand (ICOSL), IL1, Src-like adapter protein (SLA) and thrombospondin-1 (THBS1). As controls, we included several stress-response genes known to be modulated in response to UV irradiation (GDF15, FDXR, PLK2, PCNA). The RT-PCR analysis was carried out with the same RNA samples used for the microarrays. The results confirm the altered expression of all the genes tested except for AICDA and CD163 (Fig. 1).

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