In vitro biological activity of anti-C II TA hammerhead ribozyme--a novel approach for autoimmune diseases.
- Author:
Fang LIU
1
;
Ping ZOU
;
Rong GUO
;
Huazhong LU
;
Huahua FAN
Author Information
1. Institute of Hematology, Xiehe Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022.
- Publication Type:Journal Article
- MeSH:
Autoimmune Diseases;
therapy;
Base Sequence;
Genetic Therapy;
HLA-DP Antigens;
metabolism;
HLA-DQ Antigens;
metabolism;
HLA-DR Antigens;
metabolism;
HeLa Cells;
Humans;
Interferon-gamma;
pharmacology;
Molecular Sequence Data;
Nuclear Proteins;
immunology;
RNA, Catalytic;
genetics;
metabolism;
pharmacology;
physiology;
Recombinant Proteins;
Reverse Transcriptase Polymerase Chain Reaction;
Trans-Activators;
antagonists & inhibitors;
genetics;
immunology;
Transfection
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2003;23(4):335-338
- CountryChina
- Language:English
-
Abstract:
This study investigated the feasibility of using an hammerhead ribozyme against C II TA, a major regulator of MHC II antigens, to repress the expression of MHC II molecules on Hela cells. A hammerhead ribozyme (Rz464) specific to 463-465 GUC triplet of C II TA and its target gene were transcribed, then mixed up and incubated in vitro. The cleavage products were analyzed by PAGE and silver-staining. Rz464 was then inserted into the pIRES2-EGFP vector (pRz464). Stable transfectants of Hela with pRz464 were tested for class II MHC induction by recombinant human interferon-gamma (IFN-gamma). mRNA of C II TA was measured by RT-PCR. Our results showed that Rz464 could exclusively cleave C II TA RNA. When induced with IFN-gamma, the expression of HLA-DR, -DP, -DQ on pRz464+ Hela was induced, and the mRNA content of C II TA decreased too. It is concluded that Rz464 could inhibit C II TA and thus the family of genes was regulated by C II TA:MHC II molecules. These results provided insight into the future application of Rz464 as a new nucleic acid drug against auto-immune diseases.
