Silencing of SOCS1 and IL-12 gene cotransferred by adenoviral enhances DC-mediated anti-laryngocarcinoma immunity in vitro.
- Author:
Yunxia LI
1
;
Yang YUAN
;
Xuefeng WANG
Author Information
1. Liaoning Medical University, and Department of Otolaryngology, the First Affiliated Hospital of Liaoning Medical University, Jinzhou, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
Antigens, Neoplasm;
immunology;
metabolism;
Cell Line;
Cytotoxicity, Immunologic;
Dendritic Cells;
cytology;
immunology;
metabolism;
Gene Silencing;
Humans;
Interferon-gamma;
metabolism;
Interleukin-12;
genetics;
metabolism;
Laryngeal Neoplasms;
immunology;
Leukocytes, Mononuclear;
cytology;
immunology;
Lymphocyte Activation;
RNA, Small Interfering;
Suppressor of Cytokine Signaling 1 Protein;
Suppressor of Cytokine Signaling Proteins;
genetics;
metabolism;
T-Lymphocytes, Cytotoxic;
immunology;
Transfection
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2012;26(19):890-896
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects and the related immunological mechanisms of dendritic cells (DCs) modified by SOCS1siRNA gene and IL-12 gene on activating and inducing cytotoxic T lymphocyte (CTL) as well as specific immune critically killing laryngocarcinoma in vitro.
METHOD:DCs were derived from human peripheral blood mononuclear cells (hPBMC), modified by recombinant SOCSlsiRNA adenoviral and IL-12 adenoviral and then pulsed with tumor antigen of repeated freeze-thaw method. The IL-12 and IFN-y levels in culture supernatant of DCs and CTILs were examined by ELISA.
RESULT:DC were cultivated successfully and had special morphologic haracteristicistics. The rate of Ad-GFP carrying fluorescent expression was over 90%. The expression of SOCS1 protein in DCs were effectively decreased by being modified SOCSlsiRNA and IL-12 genetic while the expression of IL-12 protein were increased. The secretion rate of IL-12 factor was higher than that of SOCSlsiRNA and IL-12 transfection of single gene respectively in modified DCs which could prompt T cell proliferation activation significantly as well. IFN-y was secreted constantly in DC and CTL, resulting in Hep-2.
CONCLUSION:DC modified by SOCSlsiRNA and IL-12 gene which pulsed with laryngeal carcinoma antigen could increased the production of IL-12 and IFN-y; DC modified by SOCSlsiRNA and IL-12 gene which pulsed with laryngeal carcinoma antigen could enhance the ability to stimulate proliferation of T cell, increase production of IFN-y, IL-12 by T cells and induce the stronger killing rate of CTL.
