[Study of reduction rat T-lymphocytes activation and proliferation by inhibition inducible co-stimulator gene expression].
OBJECTIVE
To observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Liu
Wei
W
Department of Orthopedic Surgery, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Bi
Zheng-gang
ZG
Lu
Xiao-feng
XF
Zhang
Zhen-yu
ZY
Zhang
Shi-qian
SQ
Xin
Wei-guang
WG
Bo
Chao-gang
CG
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D005260:Female; D005822:Genetic Vectors; D060889:Inducible T-Cell Co-Stimulator Protein; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008213:Lymphocyte Activation; D008297:Male; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
D000945:Antigens, Differentiation, T-Lymphocyte; C556695:Icos protein, rat; D060889:Inducible T-Cell Co-Stimulator Protein; D015847:Interleukin-4; D007371:Interferon-gamma