[The biological activity of MHC classII transactivator ribonuclease P: a novel approach for hepatic transplantation rejection].
OBJECTIVE
This paper studied the effect of RNaseP against CIITA on repressing class II MHC (MHCII) expression.


METHODS
It was constructed that M1-RNA with guide sequences (GS), recognizing the 629 site of CIITA (M1-629-GS), by PCR from pTK117 plasmid, then was cloned into psNAV (psNAV-M1-629-GS). CIITA target gene was obtained from Raji cell by RT-PCR, and then inserted into pGEM-7zf (+) (pGEM-800). psNAV-M1-629-GS and pGEM-800 were transcribed and then mixed up and incubated in vitro. Stable transfectants of hepatocyte with psNAV-M1-629-GS by nanometer were tested for MHCII induction by recombinant human interferon-gamma (IFN-gamma). mRNA abundance of CIITA was measured by RT-PCR.


RESULTS
It showed that M1-629-GS could exclusively cleave pGEM-800 that formed a base pair with the GS. When induced with IFN-gamma, the expression of HLA-DR, -DP, -DQ on psNAV-M1-629-GS+ hepatocyte was (1.01+/-0.51)%, (4.37+/-1.28)%, (1.98+/-0.42)% respectively, was down-modulated 90.65%, 89.11% and 65.32% compared with control, while the mRNA content of CIITA reduced significantly (P<0.01).


CONCLUSIONS
M1-629-GS could effectively repress MHCII expressing through cleaving CIITA mRNA. These results provided insight into the future application of it as a new nucleic acid drug against the rejection of hepatic transplantation.
Guo
Rong
R
Institute of Hematology, Union Hospital, Affiliated to TongJi Medical College, HuangZhong University of Science and Technology, WuHan, China.
Zou
Ping
P
Wu
Shu-shan
SS
Cao
Yi-lin
YL
Lu
Hua-zhong
HZ
Fan
Hua-hua
HH
Gao
Feng
F
D006084:Graft Rejection / Q000517:prevention & control*; D000949:Histocompatibility Antigens Class II / Q000032:analysis*; D006801:Humans; D016031:Liver Transplantation / Q000276:immunology*; D009687:Nuclear Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D043262:Ribonuclease P / Q000494:pharmacology*; D015534:Trans-Activators / Q000235:genetics*
D000949:Histocompatibility Antigens Class II; C083728:MHC class II transactivator protein; D009687:Nuclear Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D043262:Ribonuclease P