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Chinese Journal of Surgery ; (12): 301-304, 2008.
Article in Chinese | WPRIM | ID: wpr-237797

ABSTRACT

<p><b>OBJECTIVE</b>To observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.</p><p><b>METHODS</b>Four 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.</p><p><b>RESULTS</b>After 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.</p><p><b>CONCLUSIONS</b>RNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.</p>


Subject(s)
Animals , Female , Male , Rats , Antigens, Differentiation, T-Lymphocyte , Genetics , Metabolism , Cell Proliferation , Cells, Cultured , Genetic Vectors , Inducible T-Cell Co-Stimulator Protein , Interferon-gamma , Metabolism , Interleukin-4 , Metabolism , Lymphocyte Activation , RNA Interference , Rats, Sprague-Dawley , T-Lymphocytes , Allergy and Immunology , Metabolism , Transfection
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