中国人兽共患病学报2025，Vol.41Issue(6)：559-566,8.DOI:10.3969/j.issn.1002-2694.2025.00.101
Ccdc115基因对鼠伤寒沙门菌感染RAW264.7细胞吞噬体酸化及生存能力的影响
Effects of the Ccdc115 gene on the phagosome acidification and viability of RAW264.7 cells infected with Salmonella Typhimurium
摘要
Abstract
This work was aimed at analyzing the protein characteristics of Coiled-Coil Domain-Containing Protein 115(CCDC115)and using Ccdc115-deficient mouse monocyte-macrophage leukemia cells(RAW264.7)to explore the influence of CCDC115 on the intracellular survival of Salmonella Typhimurium.Bioinformatics analysis was conducted to examine the fundamental attributes of CCDC115,which was determined to be an unstable protein consisting of two α-helices and an intervening disordered re-gion,devoid of any transmembrane structural domains.A RAW264.7-Ccdc115-KO cell line was successfully established with CRISPR/Cas9 gene-editing technology.To elucidate the effects of CCDC115 on the intracellular survival of Salmonella Typhimurium,we infected RAW264.7 cells with Salmonella Typhimurium.The expression of CCDC115 was found to be upregulated at both the mRNA and protein levels post-infection,according to RT-qPCR and western blot analysis.Via counting of colony-forming units(CFU),the proliferation rate of Salmonella Typhimurium within RAW264.7-Ccdc115-KO cells was found to be 1.5-fold higher than that in RAW264.7 cells.Acidification imaging studies indicated that,whereas Salmonella Typhimurium phagosomes underwent acidifi-cation in RAW264.7 cells,this process was absent in RAW264.7-Ccdc115-KO cells.In conclusion,the study successfully estab-lished a RAW264.7-Ccdc115-KO cell line and demonstrated that the expression of CCDC115 is elevated during Salmonella Ty-phimurium infection,thus potentially inhibiting the intracellular survival of Salmonella Typhimurium by facilitating phagosome acidifi-cation.This study lay a theoretical foundation for functional studies of CCDC115 and the investigation of mechanisms regulating the survival of intracellular Salmonella Typhimurium.关键词
卷曲螺旋结构域蛋白115/鼠伤寒沙门菌/RAW264.7/胞内生存/吞噬体酸化Key words
Coiled-Coil Domain-Containing Protein 115/Salmonella Typhimurium/RAW264.7/intracellular survival/phago-some acidification分类
医药卫生引用本文复制引用
谢荣现,程龙云,袁西露,李丽,李冰清,贾海红..Ccdc115基因对鼠伤寒沙门菌感染RAW264.7细胞吞噬体酸化及生存能力的影响[J].中国人兽共患病学报,2025,41(6):559-566,8.基金项目
国家自然科学基金项目(No.31900124,No.32170034),山东省自然科学基金(No.ZR2024QC015,No.ZR2024MC062) National Natural Science Foundation of China(No.31900124,No.32170034)and the Natural Science Foundation of Shandong Province(No.ZR2024QC015,No.ZR2024MC062) （No.31900124,No.32170034）
