Effects of B7-2 gene RNAi lentiviral vector on dendritic cells and T lymphocytes proliferation induced by dendritic cells in vitro / 中国免疫学杂志
Chinese Journal of Immunology
;
(12): 140-145, 2019.
Article
in Chinese
| WPRIM
| ID: wpr-744622
ABSTRACT
Objective:
To generate mouse B7-2 gene RNAi lentivirus and study its interference effects on B7-2 expression and T lymphocytes proliferation induced by dendritic cells.Methods:
Three sequences specific targeting B7-2 gene and one non-specific sequence were respectively synthesized, and inserted into lentiviral vector, then the recombinant vectors were sequencing. 293 T cells were co-transfected with lentiviral expression plasmid and packaging plasmids to produce recombinant lentivirus which titre was checked according to the expression level of green fluorescent protein ( GFP). Bone marrow cells from C57 BL/6 mice were isolated to differentiate into DCs at the present of GM-CSF, IL-4 and LPS for 48 h, then morphology and phenotypic was identified. DCs were infected by recombinant RNAi lentivirus and then the efficiency of infection and the expression of B7-2 on the surface of DCs were detected by flow cytometry. Effects on the proliferation of T cells were detected by co-culturing with DCs which were infected by B7-2 RNAi lentivirus and murine spleen T cells in vitro.Results:
DNA sequencing confirmed that three B7-2 RNAi and one non-specific recombinant lentiviral transfer plasmids were successfully constructed, the titer of recombinant lentivirus was ( 2-4) × 108 TU/ml. The recombinant lentivirus could effectively infect DC and inhibit the expression of B7-2. After the B7-2 recombinant lentivirus infection, the ability of DCs to stimulate the proliferation of T cells decreased obviously ( P<0. 05).Conclusion:
The lentiviral B7-2 gene RNAi vector can effectively silence the expression of B7-2 on the surface of DCs and inhibit the proliferation effect of T cells induced by DCs.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Immunology
Year:
2019
Type:
Article
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