کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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2405235 | 1103022 | 2009 | 7 صفحه PDF | دانلود رایگان |

ObjectiveTo transfect antigen presenting cells (APCs) with 4-1BB ligand DNA by attenuated Salmonella enterica serovar Typhimurium in vivo, and to observe the effects of ectogenous 4-1BBL on the immune functions of infected rats.MethodsAttenuated Salmonella typhimurium (vaccine strain) carrying plasmids pIRES2-EGFP-4-1BBL was constructed and used to infect HepG2 hepatoma cells. The expression of reporter gene, green fluorescent protein (GFP) and rat 4-1BBL in the transfected cells was detected by double-immunofluorescence staining. Rats were fed with the recombinant bacteria intragastrically on three occasions in 2 weeks, and were then sacrificed. The transcription and expression of GFP and 4-1BBL genes in splenocytes were measured by RT-PCR and flow cytometry. The phenotypes of T cells in peripheral blood and splenocytes were determined by flow cytometry. The content of IFN-γ in the cultural supernatant of splenocytes stimulated by PHA was measured by ELISA.ResultsThe recombinant bacteria harboring 4-1BBL had the same invasive abilities as the original bacteria, and it was able to deliver exogenous genes into HepG2 cells, where the GFP and 4-1BBL were successfully expressed. There were significant upregulations of CD3+CD8+ T cells (P = 0.018) and CD3+CD25+ T cells (P = 0.019) in the peripheral blood cells as well as CD3+CD8+ T cells (P = 0.022), and CD3+CD25+ T cells (P = 0.008) in splenocytes of the infected rats. The rats had more 4-1BBL expression detected in the spleen. IFN-γ released by PHA-stimulated splenocytes increased significantly by the recombinant bacteria as compared with controls (P = 0.002).ConclusionSalmonella serovar Typhimurium containing 4-1BBL can transfect target genes into antigen presenting cells in vivo, and the expression of exogenous 4-1BBL enhances cellular immunity markedly.
Journal: Vaccine - Volume 27, Issue 11, 10 March 2009, Pages 1717–1723