Construction of a recombinant plasmid harbouring the glyceraldenyde-3-phosphate dehydrogenase gene of periodic Brugia malayi and observation on DNA immunity.

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dc.contributor.authorFang, Z
dc.contributor.authorTong, H
dc.contributor.authorZhang, S
dc.contributor.authorFang, H
dc.contributor.authorLu, S
dc.contributor.authorXu, B
dc.date.accessioned2012-12-18T07:49:27Z
dc.date.available2012-12-18T07:49:27Z
dc.date.issued2012-04
dc.description.abstractPurpose: Controlling and eliminating lymphatic filariasis will require further research of preventative measures and implementation. Parasite is dependent on glycolysis for ATP production. The glycolytic enzyme glyceraldenyde-3-phosphate dehydrogenase (GAPDH) plays an important role in glycolysis and therefore is either a potential target for anti-parasite drug development or a vaccine candidate. Therefore, we tried to investigate the DNA vaccine-elicited immune responses in BALB/c mice. Materials and Methods: We cloned a gene encoding the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from periodic Brugia malayi into vector pcDNA3.1. Mice were injected at a dosage of 100 μg recombinant plasmid DNA with CpG intramuscular injection and immunized three times at 2-week intervals. pcDNA3.1 and normal saline were used as control. The tissue of muscles at the 4 weeks after the third injection was collected and target genes were detected using RT-PCR. The humoral responses elicited in mice by inoculation with the recombinant plasmid pcDNA3.1-BmGAPDH were detected using a standard ELISA. Two weeks after the third immunization, stimulation index (SI) was measured using the MTT method and the level of secreted IL-4 and INF-g were detected using ELISA. Results: Specific gene fragment coding GAPDH was amplified and the recombinant plasmid pcDNA3.1-BmGAPDH was constructed. Post-challenge sera from the mice immunized with the DNA vaccine had specific antibody titres of 1:1600 to 1:6400, and the highest titre was observed in the mice that were inoculated by pcDNA3.1-BmGAPDH/CpG at 6 weeks. At 4 weeks after immunization, the spleens of the mice were obviously enlarged. The proliferation of spleen T lymphocytes seen on the MTT assay was higher in the pcDNA3.1-BmGAPDH group than in the control group (P value <0.05). The levels of IL-4 and INF-g in serums from the immunized mice were significantly higher than that of the control (P value <0.05). Conclusions: We conclude that the recombinant eukaryotic plasmid pcDNA3.1-BmGAPDH could elicit humoral and cellular immune responses in mice.en_US
dc.identifier.citationFang Z, Tong H, Zhang S, Fang H, Lu S, Xu B. Construction of a recombinant plasmid harbouring the glyceraldenyde-3-phosphate dehydrogenase gene of periodic Brugia malayi and observation on DNA immunity. Indian Journal of Medical Microbiology. 2012 Apr-June; 30(2): 193-197.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/143944
dc.language.isoenen_US
dc.source.urihttps://www.ijmm.org/article.asp?issn=0255-0857;year=2012;volume=30;issue=2;spage=193;epage=197;aulast=Fangen_US
dc.subjectDNA vaccineen_US
dc.subjectglyceraldenyde-3-phosphate dehydrogenaseen_US
dc.subjectimmune responseen_US
dc.subjectmiceen_US
dc.subjectperiodic Brugia malayien_US
dc.subject.meshAdjuvants, Immunologic --administration & dosage
dc.subject.meshAnimals
dc.subject.meshAntibodies, Helminth --blood
dc.subject.meshBrugia malayi --enzymology
dc.subject.meshBrugia malayi --genetics
dc.subject.meshBrugia malayi --immunology
dc.subject.meshCell Proliferation
dc.subject.meshElephantiasis, Filarial --immunology
dc.subject.meshElephantiasis, Filarial --prevention & control
dc.subject.meshEnzyme-Linked Immunosorbent Assay
dc.subject.meshFemale
dc.subject.meshGlyceraldehyde-3-Phosphate Dehydrogenases --genetics
dc.subject.meshGlyceraldehyde-3-Phosphate Dehydrogenases --immunology
dc.subject.meshInjections, Intramuscular
dc.subject.meshMice
dc.subject.meshMice, Inbred BALB C
dc.subject.meshOligodeoxyribonucleotides --administration & dosage
dc.subject.meshPlasmids --administration & dosage
dc.subject.meshSpleen --immunology
dc.subject.meshT-Lymphocytes --immunology
dc.subject.meshVaccination --methods
dc.subject.meshVaccines, DNA --administration & dosage
dc.subject.meshVaccines, DNA --genetics
dc.subject.meshVaccines, DNA --immunology
dc.subject.meshVaccines, Synthetic --administration & dosage
dc.subject.meshVaccines, Synthetic --genetics
dc.subject.meshVaccines, Synthetic --immunology
dc.titleConstruction of a recombinant plasmid harbouring the glyceraldenyde-3-phosphate dehydrogenase gene of periodic Brugia malayi and observation on DNA immunity.en_US
dc.typeArticleen_US
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