Study of reduction rat T-lymphocytes activation and proliferation by inhibition inducible co-stimulator gene expression.
- Author:
Wei LIU
1
;
Zheng-gang BI
;
Xiao-feng LU
;
Zhen-yu ZHANG
;
Shi-qian ZHANG
;
Wei-guang XIN
;
Chao-gang BO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antigens, Differentiation, T-Lymphocyte; genetics; metabolism; Cell Proliferation; Cells, Cultured; Female; Genetic Vectors; Inducible T-Cell Co-Stimulator Protein; Interferon-gamma; metabolism; Interleukin-4; metabolism; Lymphocyte Activation; Male; RNA Interference; Rats; Rats, Sprague-Dawley; T-Lymphocytes; immunology; metabolism; Transfection
- From: Chinese Journal of Surgery 2008;46(4):301-304
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.
METHODSFour interference sites targeting at rat ICOS gene were designed and four pairs of oligonucleotide fragments were cloned into the pSilencer 4.1-CMV neo plasmid vectors then transfected into rat lymphocytes with cationic liposome. The expression of mRNA and protein of ICOS was detected by RT-PCR and flow cytometry. The alteration of lymphocyte proliferation ability was evaluated by mix lymphocyte reaction, and the secretion levels of IFN-gamma and IL-4 were measured by ELISA procedure.
RESULTSAfter transfection, the expression of mRNA and protein of ICOS in test groups were lower than that in control groups (P < 0.05). The ability of T-lymphocytes in proliferation was poor and the levels of IFN-gamma and IL-4 were reduced with ICOS gene shut down.
CONCLUSIONSRNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.
