The Construction of Human Cytomegalovirus pp65 Prokaryotic Expressing Vector and the Inducement of Specific Immune Response / 中华围产医学杂志

ABSTRACT

Objective To construct human cytomegalovirus pp65 prokaryotic expressing vector and induce specific T lymphocyte immune response with recombinant pp 65 protein of human cytomegalovirus(HCMV) to observe the effect of CTL on infected HEL cells. Methods The whole cDNA of HCMV pp65 was amplified by PCR and inserted into prokaryotic expressing vector pRSET by gene engineer technique. The product was purified by affinity chromphotography and identified with Western blot after that recombinant plasmid was expressed by the induction of IPTG. With MTT technique, we observed the stimulating and proliferating effect of recombinant HCMV pp65 protein on PBMC in vitro. Cytotoxicity of PBMC on HCMV-infected HEL cells was detected. Results The pp65 prokaryotic expressing vector was successfully constructed and could express in engineering bacteria DE3. High dose of pure recombinant protein was acquired and had been identified. The rhHCMV pp65 protein can activate PBMC and cause the proliferation of it in vitro. The proliferated PBMC have the specific cytotoxicity to HEL cells infected by HCMV. Conclusions The acquired recombinant HCMV pp65 protein could induce specific T cells immune response in vitro to kill the HCMV infected HEL cells. And it is very important for the immune therapy of the HCMV infections.