Recombinant Expression, Antibody Preparation And Clinical Application Of Human Granulysin

To obtain biological-activited GNLY protein and study on the relationship between GNLY and some disease,we successfully constructed the recombinant expression plasmid pET28a(+)-GNLY and obtained the purified recombinant fusion protein of 9kD GNLY that was expressed in E.coli. It was approved that this protein has immunogenicity by Western Blot and can kill the tumor cells by MTT.GNLY was used to immunized rabbit and we obtained high titers polyclonal anti-GNLY antibody. ELISA was established to detect GNLY in the serum.The results showed that GNLY in hepatocellular carcinoma patients were lower than those in the normal cohort (p<0.05) ,while GNLY in acute phase HBV infection patients were significantly higher than those in the normal cohort (p<0.01) .We successfully established the FQ-PCR to detect GNLY mRNA of PBMC. The results showed that the mRNA levels of GNLY in hepatocellular carcinoma patients were lower than those in the normal cohort (p<0.05) ,while the levels in acute phase HBV infection patients showed significantly higher than those in the normal cohort (p<0.01) .These intriguing preliminary results suggested that GNLY may be associated with immunity state of infection diseases and some carcinomas.