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Archive > Volume 24 Issue 9 > 2017,24(9):950-954. DOI:10.3872/j.issn.1007-385X.2017.09.004 Prev Next
Basic Research
Effect of TLR-4 on phenotypes and function of dendritic cells
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Abstract:
Objective: To investigate whether the expression of Toll like receptor-4 (TLR-4) has an effect on the phenotypes and function of dendritic cells (DCs) and the possible mechanism. Methods: pcDNA3.1 plasmid encod-ing full TLR-4 gene sequence was constructed as a template to obtain TLR-4 mRNA through in vitro transcription us-ing mMESSAGE mMACHINE T7 Kit; DCs from healthy human peripheral blood were transfected with TLR-4 mRNA by liposomal transfection. The functional molecule expression on DC surface and the ability of DC inducing cytotoxic T lymphocytes (CTLs) to secrete cytokines in vitro were detected by Flow cytometry after transfection.Results: Human TLR-4-pcDNA3.1 plasmid was successfully constructed; human TLR-4 and EGFP mRNA frag-ments were successfully amplified. The Flow cytometry results showed that the expressions of chemokine CCR7 and functional molecule HLA-DR on dendritic cell surface were significantly increased after transfection with TLR-4 mRNA compared with control mRNA transfection or pre-ransfection (CCR7: [42.4±4.93]% vs [20.1±3.09]%，[17.1±4.33]%，P<0.05; HLA-DR:[62.1±7.23]% vs [17.7±6.01]%，25.8±4.16]%，P<0.05); and the ability of secret-ing IFN-γ from CTLs induced by TLR-4 mRNA-DCS was significantly enhanced in vitro compared with control mRNA-DC，empty-DC and PBMC ([66.5±3.58]% vs [41.1±4.27]%，[37.9±2.96]%，[3.2±2.03]%，P<0.05). Conclu-sion: The function of dendritic cells could be significantly enhanced by TLR-4 mRNAtransfection. The ability of an-tigen presentation and inducing CTLs of dendritic cell were improved, which would provide an experimental basis for enhancing the anti-tumor effect of DC vaccines.
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Project Supported:
Project supported by the National Clinical Key Specialty Construction Program，the Fujian Science and Technology Plan （No.2017Y0022），and the Natural Sci-ence Foundation of Fujian Province （No.2016J01514）
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