为研究人源福氏志贺菌对鸡肠上皮原代细胞侵袭力和侵袭特性,进一步验证人源志贺菌对鸡的致病性并建立体外研究模型,采用Hela细胞庆大霉素保护试验和免疫组织化学染色检测分离的人源福氏志贺菌ZD02株的毒力.然后采用细胞免疫组织化学染色研究ZD02株对鸡肠上皮细胞的侵袭力和相关侵袭特性.人源福氏志贺菌ZD02株能够侵入Hela细胞,庆大霉素保护试验结果显示,感染后30、60、90、120 min时ZD02株侵袭率分别为1.43％、2.43％、2.56％、2.62％,证明ZD02株具有毒力.ZD02株侵袭鸡肠上皮原代细胞免疫组织化学检测结果显示,ZD02株能侵入鸡肠上皮细胞,感染后1、2、3、4h时ZD02株侵袭率分别为1.7％、6.2％、8.0％、11.2％.鸡肠上皮原代细胞经EGTA处理后,ZD02株侵袭率显著提高.人源福氏志贺菌ZD02株对鸡肠上皮细胞具有侵袭力,且从肠上皮细胞基底侧部侵袭.本研究进一步验证了志贺菌人禽互传的可能性,鸡肠上皮原代细胞可作为研究志贺菌致病机理的体外模型.%In order to further determine pathogenicity of human source S. flexneri to chicken, invasion and invasive characteristics of human source S. flexneri ZD02 strain on primary chicken intestinal epithelial cells were studied, and an in vitro model was established. Virulence of the isolated human source S. flexneri ZD02 strain was validated by invasion of Hela cell using Gen-tamicin resistance assay and immunohistochemistry. Invasion and invasive characteristics of ZD02 strain on primary chicken intestinal epithelial cells were researched by immunohistochemistry. Human source S. flexneri ZD02 strain could invade into Hela cells, and the invasion ratios were 1. 43%, 2. 43%, 2. 56%, 2. 72% at 30, 60, 90, 120 minutes after infection respectively. The result of immunohistochemistry showed that ZD02 strain could invade into primary chicken intestinal epithelial cells, and the invasion ratios were 1. 7%, 6. 2%, 8. 0%, 11. 2% at 1, 2, 3, 4 hours after infection respectively. Primary chicken intestinal epithelial cells treated with EGTA which disrupts intercellular junctions, significantly enhanced invasion ratio of ZD02 strain. Human source S. flexneri ZD02 strain can invade into primary chicken intestinal epithelial cells from the basolateral pole of cells, which further validated the possibility of mutual infection caused by S. flexneri between human and avian. Primary chicken intestinal epithelial cells can be used as an in vitro model to study the pathogenic mechanism of Shigella.
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