Establishment of a leptospirosis model in guinea pigs using an epicutaneous inoculations route.
Establishment of a leptospirosis model in guinea pigs using an epicutaneous inoculations route.
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经皮接种途径建立豚鼠钩端螺旋体病模型
DOI：
10.1186/1471-2334-12-20
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发表时间：
2012-01-25
影响因子：
3.7
通讯作者：
Jiang XC
中科院分区：
文献类型：
--
作者：
Zhang Y;Lou XL;Yang HL;Guo XK;Zhang XY;He P;Jiang XC
背景推测钩端螺旋体是通过皮肤的小擦伤或破损进入宿主的。腹膜内途径虽然常用于钩端螺旋体病的豚鼠和仓鼠模型，但并不反映自然感染期间遇到的情况。本研究的目的是通过皮肤途径建立一种新型钩端螺旋体病豚鼠模型，并通过比较其他使用不同感染途径的研究数据来阐明实验豚鼠钩端螺旋体病的发病机制。方法将5×108问号钩端螺旋体赖菌株接种到豚鼠的仅剃毛或擦伤的皮肤上。在感染后2、8、24、48、72、96和144小时处死豚鼠，然后收获肺、肝、肾、脾和接种部位周围的皮肤以供进一步检查。采用苏木精和伊红（HE）染色和电镜检测病理变化。实时PCR和免疫组化染色分别检测钩端螺旋体在血液和组织中的动态分布。结果在擦伤皮肤接种的豚鼠中，早在感染后2小时（pi.i.）血液中就检测到了钩端螺旋体，然后在感染后96小时内传播到几乎所有动物的肝脏、肺和肾脏。在肿胀中也检测到了钩端螺旋体被吞噬。血管内皮细胞经常聚集在接种部位下方真皮和皮下组织的毛细血管周围。皮肤擦伤接种的豚鼠，在内脏器官出血出现之前，已发现接种部位周围真皮出血，还出现肺部出血、肾炎、黄疸、血尿等严重病变，7只豚鼠于接种后144小时死亡2只。而使用相同剂量Lai菌株的仅剃毛的豚鼠中未检测到病变和钩端螺旋体。结论完整的角质形成细胞层是对抗钩端螺旋体的非常有效的屏障，完整的皮肤可以阻止钩端螺旋体向宿主的浸润。钩端螺旋体可以穿透擦伤的皮肤，并通过组织屏障迅速建立全身感染。我们通过皮下接种途径成功建立了一种新型钩端螺旋体病豚鼠模型，该模型复制了自然感染过程，似乎是研究钩端螺旋体病发病机制的另一种方法，特别是在宿主与病原体相互作用的早期阶段。这种新模型也可能有利于研究皮肤屏障和表皮与该生物体相互作用的机制。
BackgroundLeptospires are presumed to enter their host via small abrasions or breaches of the skin. The intraperitoneal route, although commonly used in guinea pig and hamster models of leptospirosis, does not reflect conditions encountered during natural infection. The aim of this study is to develop a novel leptospirosis guinea pig model through epicutaneous route and to elucidate the pathogenesis of leptospirosis in experimental guinea pigs by comparing the data from other studies using different infection routes.MethodsThe guinea pigs were inoculated with 5 × 108Leptospira interrogansstrain Lai onto either shaved-only or abraded skin. The guinea pigs were sacrificed at 2, 8, 24, 48, 72, 96 and 144 h post-infection (p.i.) followed by harvest of the lungs, liver, kidneys, spleen, and the skin around the inoculated sites for further examinations. Hematoxylin and eosin (HE) staining and electron microscopy were used to detect the pathologic changes. Real time PCR and immunohistochemistry staining were performed to detect dynamic distribution of leptospires in blood and tissues, respectively.ResultsIn the guinea pigs with abraded skin inoculations, leptospires were detected in blood as early as 2 h post infection (p.i.) and then disseminated to the liver, lungs and kidneys of almost all animals within 96 h p.i.. Leptospires were also detected engulfed in the swelling vascular endothelial cells and were frequently aggregated around the capillaries in the dermis and subcutaneous tissue under the inoculated site. For the guinea pigs with abraded skin inoculations, hemorrhage at the dermis around the inoculated site was found before the appearance of internal organs hemorrhage, severe lesions such as hemorrhages in the lungs, nephritis, jaundice, haematuria were also observed, and two of seven guinea pigs died at 144 h p.i. while no lesions and leptospires were detected in the shaved-only guinea pigs using the same dose of strain Lai.ConclusionIntact keratinocyte layer is a very efficient barrier against leptospires, and intact skin can prevent the infiltration of leptosipres to the host. Leptospires can penetrate abraded skin and quickly establish a systemic infection by crossing tissue barriers. We have successfully established a novel leptospirosis guinea pig model through epicutaneous inoculations route, which replicates a natural course of infection and appears to be an alternative way to investigate the pathogenesis of leptospirosis, especially in terms of early stage of host-pathogen interactions. This novel model may also be advantageous for studies of the mechanisms involved in cutaneous barriers and epidermal interactions with this organism.
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影响因子：
5.4
作者：
Liu, Yunying;Zheng, Wei;Yan, Jie
通讯作者：
Yan, Jie
影响因子：
3.7
作者：
Lin YP;McDonough SP;Sharma Y;Chang YF
通讯作者：
Chang YF
影响因子：
3.2
作者：
JOHNSON, RC;HARRIS, VG
通讯作者：
HARRIS, VG
影响因子：
3.1
作者：
Hoke, David E.;Egan, Suhelen;Adler, Ben
通讯作者：
Adler, Ben
影响因子：
4.2
作者：
Atzingen MV;Barbosa AS;De Brito T;Vasconcellos SA;de Morais ZM;Lima DM;Abreu PA;Nascimento AL
通讯作者：
Nascimento AL