Recombinant attenuated Salmonella harboring 4-1BB ligand gene enhances cellular immunity
Recombinant attenuated Salmonella harboring 4-1BB ligand gene enhances cellular immunity
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DOI：
10.1016/j.vaccine.2009.01.031
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发表时间：
2009-03-10
期刊：
影响因子：
5.5
通讯作者：
Chen, Wei-chang
中科院分区：
文献类型：
--
作者：
Ye, Jian-xin;Zhang, Yuan-ting;Chen, Wei-chang
目的：目的：利用减毒鼠伤寒沙门氏菌（Salmonella enterica serovar Typhimurium）体内转染含4- 1BB配体DNA的抗原提呈细胞（APC），观察外源性4- 1BBL对感染大鼠免疫功能的影响。免疫荧光双染法检测转染细胞中报告基因、绿色荧光蛋白（GFP）和大鼠4-1BBL的表达。在2周内三次用重组细菌灌胃喂养大鼠，然后处死。用RT-PCR和流式细胞术检测GFP和4-1BBL基因在脾细胞中的转录和表达。用流式细胞仪检测外周血和脾细胞中T细胞的表型。结果：4-1BBL重组菌具有与原菌相同的侵袭能力，并能将外源基因导入HepG 2细胞中，GFP和4-BBL在HepG 2细胞中得到了成功表达。感染组外周血CD 3（+）CD 8（+）T细胞（P = 0.018）和CD 3（+）CD 8（+）T细胞（P = 0.019）以及脾细胞CD 3（+）CD 8（+）T细胞（P = 0.022）和CD 3（+）CD 8（+）T细胞（P = 0.008）均显著上调。大鼠脾脏中检测到更多的4-1BBL表达。结论：含4-1 BBL的鼠伤寒沙门氏菌在体内可将目的基因导入抗原提呈细胞，外源性4- 1 BBL的表达可显著增强细胞免疫功能。递呈细胞（C）2009 Elsevier Ltd.保留所有权利。
Objective: To 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.Methods: Attenuated Salmonella typhimurium (vaccine strain) carrying plasmids pIRES2-EGFP-4-1 BBL 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-gamma in the cultural supernatant of splenocytes stimulated by PHA was measured by ELISA.Results: The 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-BBL were successfully expressed. There were significant upregulations of CD3(+)CD8(+) T cells (P = 0.018) and CD3(+)CD8(+) T cells (P = 0.019) in the peripheral blood cells as well as CD3(+)CD8(+) T cells (P = 0.022), and CD3(+)CD8(+) T cells (P = 0.008) in splenocytes of the infected rats. The rats had more 4-1BBL expression detected in the spleen. IFN-gamma released by PHA-stimulated splenocytes increased significantly by the recombinant bacter as compared with controls (P = 0.002).Conclusion: Salmonella serovar Typhimurium containing 4-1 BBL can transfect target genes into antigen presenting cells in vivo, and the expression of exogenous 4-1 BBL enhances cellular immunity markedly. presenting cells (C) 2009 Elsevier Ltd. All rights reserved.