Objective　To construct a recombinant plasmid pcDNA3-sHLA-G1 expressing soluble HLA-G1. Methods　 Total cell RNA was extracted from the cell line Jeg-3 and the cDNA was amplified by RT-PCR； The cDNA fragment was inserted into the eukaryotic expressing vector pcDNA3 and the recombinant plasmid was identified by restriction endonucleases digestion and sequencing. Results　 After restriction endonucleases treatment and sequencing, it was confirmed that the pcDNA3-sHLA-G1 had been constructed successfully. Conclusion　 In this study, the recombinant plasmid pcDNA3-sHLA-G1 had been constructed successfully.

Objective To investigate the effect of portal hypertension (PH) on the activity of local renin angiotensin system(LRAS). Method The expression of local renin and local angiotensinogen mRNA in liver,splenic artery and vein of PH patients was detected with RT-PCR. Result Expression of local renin mRNA in the liver in control group was 0.19?0.12, significantly lower than that of splenic artery (0.45?0.12) and splenic vein (0.39?0.12) respectively(P

Objective:To investigate the relationship between the level of Bcl 2 protein in EB virus infected cells and the sensitivity of this cells to NK activity and apoptosis inducing factors.Methods:Antisense oligodexynucleotides(ODNs) were used to modulate the expression of Bcl 2 gene in Epstein Barr virus transformed B lymphoblastoid cell lines(EBV LCLs);then the change of cytotoxicity of natural killer cells and the sensitivity of apoptosis inducing elements(taking out growth factor ?dexamethasone)targeting EBV LCLs,were investigated.Results:The level of Bcl 2 expression in EBV LCLs has negative relativity with the cytotoxicity of NK cells to EBV LCLs(P

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.
Animals ; CD28 Antigens ; genetics ; immunology ; Electroporation ; Humans ; Immunity, Cellular ; Interleukin-2 ; immunology ; K562 Cells ; Mice ; Muromonab-CD3 ; immunology ; Neoplasms, Experimental ; genetics ; immunology ; pathology ; RNA, Messenger ; genetics ; immunology ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; genetics ; immunology