Research of SOCS1 silent DC vaccine on laryngocarcinoma therapy.
10.13201/j.issn.1001-1781.2012.04.006
- Author:
Yang YUAN
1
;
Xuefeng WANG
;
Yang ZHANG
;
Jiayu WANG
;
Ning LUAN
Author Information
1. Department of Otolaryngology, Affiliated Hospital of Weifang Medical University, Weifang, 261031, China.
- Publication Type:Journal Article
- MeSH:
Cancer Vaccines;
immunology;
Cell Line, Tumor;
Cells, Cultured;
Dendritic Cells;
drug effects;
immunology;
Gene Silencing;
Granulocyte-Macrophage Colony-Stimulating Factor;
pharmacology;
Humans;
Interleukin-4;
pharmacology;
Laryngeal Neoplasms;
therapy;
RNA, Small Interfering;
Suppressor of Cytokine Signaling 1 Protein;
Suppressor of Cytokine Signaling Proteins;
genetics;
Tumor Necrosis Factor-alpha;
pharmacology
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2012;26(4):169-173
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:Investigate the specific antitumor mechanism of SOCS1 silent DC vaccine and discuss the prospect of RNAi in the gene therapy for laryngocarcinoma in order to provide novel ideas of DCs clinical applications.
METHOD:Dendritic cells derived from peripheral blood monocytes were cultured in vitro in the presence of GM-CSF, IL-4 and TNF-alpha. The morphological feature of DC was observed with inverted microscope. RNAi vector were transfected into DC. The expression of SOCS1 protein was detected with Western blot. The effective target sequences of siRNA against SOCS1 were screened out. The surface markers of mature DC, including CD83, CD86 and HLA-DR, were detected with flow cytometry. The concentration of IFN-gamma in the supernatant was assayed by ELISA. The proliferative ability of T cell stimulated by DC and the specific killing activity of cytotoxic T lymphocyte (CTL) induced by DC were evaluated by MTT assay.
RESULT:Dendritic cells were obtained successfully. The RNAi vector was proved to be right by sequencing. The expression of SOCS1 decreased significantly under the influence of the 5th interference sequence. SOCS1 silent dendritic cells which were loaded with Hep-2 antigen had high expressions of CD83 (85.61 +/- 0.96)%, CD86 (96.86 +/- 1.20)% and HLA-DR (98.02 +/- 0.94)%. It could also stimulate the proliferation of T cells effectively as well as could increase the production of IFN-gamma, eventually enhanced the specific killing effect of CTL. The killing activity was more higher than that in control group when the effect cells and target cells were mixed up at the ratio of 50:1 (P < 0.01).
CONCLUSION:SOCS1 silent DC vaccines which were loaded with Hep-2 antigen could induce effective and specific anti-laryngocarcinoma immune responses.
