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In vitro maturation and migration of immature dendritic cells after chemokine receptor 7 transfection

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Abstract:

Dendritic cells are specialized antigen-presenting cells that regulate immunity and tolerance. Chemokine receptor 7 (CCR7), which is expressed by mature dendritic cells, mediates the migration of the cells to secondary lymphoid organs and thus regulates immune responses. It has been demonstrated that immature dendritic cells can induce immune tolerance, but they do not express CCR7 and cannot migrate to secondary lymphoid organs. We transfected immature dendritic cells with a recombinant adenovirus carrying the CCR7 gene to obtain immature dendritic cells with the ability to migrate. The maturity of the cells was monitored by scanning electron microscopy and flow cytometry. In addition, we assessed the ability of cells to migrate and the function of the cells using in vitro chemotactic and mixed leukocyte reaction assays. The results showed that immature dendritic cells became semi-mature, exhibiting a mild upregulation of co-stimulatory molecular expression and a few dendritic processes. Immunofluorescence assay and Western blotting indicated that CCR7 protein expression increased significantly in immature dendritic cells following CCR7 gene transfection. The in vitro chemotactic assay showed a significantly enhanced ability to migrate in response to CCL19 following CCR7 gene transfection. Moreover, transfected cells showed an enhanced ability to stimulate allogeneic T cell proliferation in vitro, but their ability was significantly weaker than that of mature dendritic cells. Interleukin-10 inhibited the differentiation and maturation of immature dendritic cells. It is concluded that, following CCR7 gene transfection, immature dendritic cells exhibit an enhanced ability to migrate and some of the characteristics of mature cells. Thus, these cells are of potential clinical significance in studies of immune tolerance induction during skin grafting after severe burns.

Les cellules dendritiques sont des cellules présentatrices d’antigènes spécialisées qui régulent l’immunité et la tolérance. Le récepteur de chimiokines 7 (CCR7), exprimé sur les cellules dendritiques matures, permet dans la migration des cellules vers les organes lymphoïdes secondaires et régule ainsi les réponses immunes. Les cellules dendritiques immatures peuvent induire la tolérance immunitaire mais elles n’expriment pas le CCR7 et ne peuvent pas migrer vers les organes lymphoïdes secondaires. Nous avons transfecté des cellules dendritiques immatures avec un adénovirus recombinant comportant le gène du CCR7 afin d’obtenir des cellules immatures ayant la propriété de migrer. La maturité des cellules a été examinée par microscopie électronique à balayage et par cytométrie de flux. En plus, nous avons évalué la capacité de migration des cellules et les fonctions cellulaires par des essais de chimiotaxie in vitro et en culture mixte de leucocytes. Les résultats ont montré que les cellules dendritiques immatures devenaient semi-matures, démontrant une faible augmentation de l’expression des molécules de co-stimulation et de quelques processus dendritiques. Des essais en immunofluorescence et en immunobuvardage ont indiqué que l’expression du CCR7 augmentait significativement chez les cellules dendritiques après la transfection du gène du CCR7. L’essai de chimiotaxie in vitro a montré une augmentation significative de la capacité de stimuler la prolifération de cellules T allogéniques in vitro, mais leur capacité était significativement plus faible que celles des cellules dendritiques matures. L’IL-10 inhibait la différenciation et la maturation des cellules dendritiques immatures. Nous concluons que suite à la tranfection du gène du CCR7, les cellules dendritiques démontrent une plus grande capacité de migration et quelques caractéristiques de cellules matures. Ainsi, ces cellules possèdent un potentiel clinique intéressant notamment pour des études d’induction de la tolérance immune lors de greffes de peaux après des brûlures sévères.

Document Type: Research Article

Publication date: 2009-07-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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