The Immunosupression After Liver Transplantation Induced By Recipient Lymphocytes Carrying ShRNA Against IL2 Gene

PART ONE CONSTRUCTION AND IDENTIFICATION OF EXPRESSING SHRNA AGAINST RAT IL-2Objective: To successfully develop a king of RNA interference(RNAi) that specifically targets rat interleukin-2 by synthesizing short hairpin RNA(shRNA) in vivo, and to assess the inhibitory effect of this shRNA on the expression of IL-2 mRNA in rat activivated T lymphocytes.Methods: Two different recombinant plasmid ( pIL-2A, pIL-2B ) targeting rat IL-2 gene sequence and their unrelated control plasmid pIL-2C were constructede based on pGensil-2 vector, and were identified with digestion and sequencing. These recombinant plasmids were transfected into freshly isolated rat lymphocytes with cationic liposome. At 24h post transfection, the production of IL-2 in supernatant is measured by enzyme linked immunosorbent assay (ELISA), and the change of the IL-2 mRNA levels was detected with reverse transcription-polymerase chain reaction (RT-PCR).Results: These recombinant plasmids were successlly constructed and identified with digestion and sequencing. At 24h post transfection , the results of ELISA and semi-quantitative RT-PCR assay indicated that these...