Allo-cell transplant rejection is associated with class II major histocompatibility complex (MHC II), while its transactivator (namely C II TA) regulates MHC II molecules expression strictly and exclusively. The aim of this study was to investigate the inhibiting effect of C II TA anti-sense RNA on MHC II expression. The cDNA for anti-sense RNA recognizing the 114-523 sequence of C II TA (arC II TA) was obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid (pcDNA3.1B-arC II TA, pD-arC II TA). Raji cells were transfected stably with pD-arC II TA, classic MHC II antigen (HLA-DR, -DP, -DQ) expression was assayed by flow cytometry (FCM). mRNA abundance of C II TA, invariant chain and classic MHC II were detected by RT-PCR. The results showed that compared with control (sense C II TA), the expression inhibition of HLA-DR, -DP, -DQ on pD-arC II TA positive Raji cell was 65.93%, 54.14%, 68.32% respectively. The mRNA contents of C II TA, invariant chain and classic MHC II also decreased. In conclusion, arC II TA inhibited C II TA and thus the family of MHC II molecules were regulated by it, therefore these results provide a novel approach for the control of graft versus host diseases.
Cell Line, Tumor ; Flow Cytometry ; Graft Rejection ; genetics ; prevention & control ; HLA-DP Antigens ; biosynthesis ; genetics ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Nuclear Proteins ; genetics ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection