Suppressor of cytokine signaling 1 (SOCS1) silencing and Hep-2 sensitizing dendritic cell vaccine in laryngocarcinoma immunotherapy

Y. Yuan, G.-Y. Li, M. Ji, Y. Zhang, Y.-P. Ding, X.-C. Qi

Department of Otolaryngology-Head and Neck Surgery, Affiliated Hospital of Weifang Medical University, Weifang, Shandong, P.R. China. qhy302@163.com

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• Pharmacology

OBJECTIVE: Many studies have recently suggested that dendritic cell (DC) vaccine contributes to the immunotherapy of various types of human tumors. It has been proved that the tumor antigen sensitizing and the gene silencing are effective methods for the preparation of the DC vaccines. The aim of this study is to investigate the specific anti-laryngocarcinoma immune response for the suppression of cytokine signaling1 (SOCS1) silencing and Hep-2 sensitizing DC.

MATERIALS AND METHODS: The dendritic cells derived from peripheral blood mononuclear cells were induced by cytokines GM-CSF, IL-4, and TNF-α in vitro, and the morphological characteristics of dendritic cells were observed under a microscope, indicating that they successfully differentiated into dendritic cells. The RNA interference vector was used to transfect dendritic cells. The expression of SOCS1 was detected by Western blot and the effective target sequence for inhibiting the expression of SOCS1 was screened. The expressions of CD83, CD86, and HLA-DR on dendritic cells were detected by flow cytometry. The content of IFN-γ in the supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). Methyl thiazolyl tetrazolium (MTT) was used to evaluate the ability of dendritic cells to stimulate T cell proliferation and induce the killing activity of cytotoxic T cells.

RESULTS: The result of PCR and Western blot analysis shows that the expression of SOCS1 significantly decreased under the influence of the 5th interference sequence. The flow cytometric analysis results show that SOCS1 silencing and Hep-2 sensitizing dendritic cells had high expressions of CD83 (85.61±0.96)%, CD86 (96.86±1.20)%, and HLA-DR (98.02±0.94)%. The DC vaccine could increase the production of IFN-γ according to the ELISA assay results. The MTT assay results show that the DC vaccine could also stimulate the proliferation of the T cells and effectively and eventually enhance the specific killing effect of CTL.

CONCLUSIONS: SOCS1 silencing and Hep-2 sensitizing DC vaccine could induce an effective and specific anti-laryngocarcinoma immune response.

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