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Tolerance Induction by CD40 Silenced Dendritic Cells through Antisense

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Author(s): MH Karimi | P Ebadi | AA Pourfathollah | ZS Soheili | SH Samiee | Z Ataee | SZ Tabei | F Nadali | SM Moazzeni

Journal: Iranian Red Crescent Medical Journal
ISSN 1561-4395

Volume: 11;
Issue: 3;
Start page: 286;
Date: 2009;
Original page

Keywords: Dendritic cell | Antisense | CD40 | Tolerance induction

ABSTRACT
Background: One of the valuable tools for inhibiting the specific gene expression is antisense technique. Todetermine T cell responses, co-stimulatory molecule expression on the antigen presenting cells is important. Inthe present study, the effects of high affinity antisense against CD40 mRNA on the function and phenotype ofDCs (dendritic cells) were investigated.Methods: The DCs were separated from the mice spleens and then cultured in vitro. By means of lipofectamine2000, the antisense was delivered into the cells and the efficacy of transfection was estimated by flow cytometry.Also, the mRNA expression and protein synthesis were assessed by real time PCR and flow cytometry, respectively.The DCs were transfected with 6 μM antisense and 2 μl lipofectamine 2000.Results: The percentage of CD40 expression in DCs was 38%. The results showed that CD40 expression isreduced in DCs to 22% and 24%. By annexine V and propidium iodine staining, we could evaluate the viability ofthe transfected cells. The inhibition of CD40 gene expression was associated with the increase in IL-4 secretion.This shifted the DCs to stimulate Th2 cytokine production from the allogenic T cells. In addition, in the MLR, theDCs without CD40 expression showed poor allostimulatory effects. This finding is valuable in the study of the costimulatorymolecules of DCs.Conclusion: These data demonstrate that direct interference of the cell surface expression of CD40 at transcriptionallevel by antisense confers tolerogenecity potential of DCs. This approach is a useful tool through whichDCs become tolerogenic and can be studied as a potential therapeutic option for the autoimmune diseases andallograft rejection.
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