The biological activity of MHC classII transactivator ribonuclease P: a novel approach for hepatic transplantation rejection.
- Author:
Rong GUO
1
;
Ping ZOU
;
Shu-shan WU
;
Yi-lin CAO
;
Hua-zhong LU
;
Hua-hua FAN
;
Feng GAO
Author Information
- Publication Type:Journal Article
- MeSH: Graft Rejection; prevention & control; Histocompatibility Antigens Class II; analysis; Humans; Liver Transplantation; immunology; Nuclear Proteins; genetics; RNA, Messenger; analysis; Ribonuclease P; pharmacology; Trans-Activators; genetics
- From: Chinese Journal of Hepatology 2003;11(12):745-748
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThis paper studied the effect of RNaseP against CIITA on repressing class II MHC (MHCII) expression.
METHODSIt 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.
RESULTSIt 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).
CONCLUSIONM1-629-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.
