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- W2373932783 abstract "Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-γwas assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-γlevel in pcD NA3.1 (+) immunized group was (23.79±11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71±30.52)pg/mL (P0.01).The stimulation index (SI) of the splenocyte proliferation was 1.05±0.30和1.94±0.74(P0.01) in pcDNA3.1(+) and pcDNA3.1(+)/NspA immunized group,respectively.The NspA gene could be detected in the muscle cells.Conclusions NspA gene vaccine is successfully constructed,which can induce specific humoral and cellular immune responses in immunized BALB/c mice model.These findings will be beneficial to the prevention of gonorrhea." @default.
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- W2373932783 date "2007-01-01" @default.
- W2373932783 modified "2023-09-25" @default.
- W2373932783 title "Construction of Neisseria surface protein A gene vaccine of Neisseria gonorrhoeae and evaluation of the immune responses induced by this vaccine in mice model" @default.
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