Combination of R848 and Poly(I: C) promotes antigen presentation by DC and enhances killing effect of CTL on lung adenocarcinoma A549 cells |
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| Authors: | HUANG Xue WANG Ce CHEN Shang YANG Xiao-fei ZHENG Yuan-yuan WANG Jing-bo LI Fu-rong |
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| Institution: | The Second Clinical Medical College of Jinan University, Shenzhen People's Hospital, Translational Medicine Collaborative Innovation Center, Shenzhen Immune Cell Therapy Public Service Platform, Shenzhen 518020, China |
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| Abstract: | AIM: To investigate the effect of R848 (a Toll-like receptor 7/8 agonist) combined with poly-inosinic:polycytidylic acidPoly(I:C), a Toll-like receptor 3 agonist] on dendritic cell (DC) maturation, and the killing effect of DC-induced cytotoxic T-lymphocytes (CTL) on human lung adenocarcinoma A549 cells. METHODS: Mononuclear cells were isolated from human peripheral blood and induced to differentiate into DC. The whole-cell lysate of A549 cells, namely tumor cell lysate (TCL), was used as antigen. R848 combined with Poly(I:C) was used as adjuvant to stimulate the DC. DC surface markers were analyzed by flow cytometry. The DC stimulated by antigen was co-cultured with T-lymphocytes for 7 d to induce CTL. The culture supernatant and CTL were collected. The levels of interleukin-12 (IL-12) p70, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant were measured by ELISA. The CTL and A549 cells were co-cultured for 16 h, and the cytotoxicity was observed by LDH assay.RESULTS: The expression of CD83 and CD80 on the DC surface, and the secretion of IL-12 p70 in DC-R848+Poly(I:C) group were significantly increased compared with DC-TCL group (P<0.01). In addition, the cytotoxicity of CTL for A549 cells in DC-R848+Poly(I:C) group was significantly enhanced compared with DC-TCL group (P<0.01). The secretion levels of IFN-γ and TNF-α in DC-R848+Poly(I:C) group were significantly elevated compared with DC-TCL group (P<0.01). CONCLUSION: R848 combined with Poly(I:C) significantly promotes DC maturation and activation, and enhances the antigen-presenting effect of DC and the cytotoxicity of DC-induced CTL. |
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| Keywords: | Dendritic cells Cytotoxic T-lymphocytes Lung adenocarcinama Toll-like receptor agonists |
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