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Ex vivo infection of murine precision-cut lung tissue slices with Mycobacterium abscessus: a model to study antimycobacterial agents
Annals of Clinical Microbiology and Antimicrobials ( IF 5.7 ) Pub Date : 2020-11-22 , DOI: 10.1186/s12941-020-00399-3
Carmen Amelia Molina-Torres 1 , Oscar Noé Flores-Castillo 2 , Irma Edith Carranza-Torres 2, 3 , Nancy Elena Guzmán-Delgado 4 , Ezequiel Viveros-Valdez 2 , Lucio Vera-Cabrera 1 , Jorge Ocampo-Candiani 1 , Julia Verde-Star 2 , Jorge Castro-Garza 3 , Pilar Carranza-Rosales 3
Affiliation  

Multidrug-resistant infections due to Mycobacterium abscessus often require complex and prolonged regimens for treatment. Here, we report the evaluation of a new ex vivo antimicrobial susceptibility testing model using organotypic cultures of murine precision-cut lung slices, an experimental model in which metabolic activity, and all the usual cell types of the organ are found while the tissue architecture and the interactions between the different cells are maintained. Precision cut lung slices (PCLS) were prepared from the lungs of wild type BALB/c mice using the Krumdieck® tissue slicer. Lung tissue slices were ex vivo infected with the virulent M. abscessus strain L948. Then, we tested the antimicrobial activity of two drugs: imipenem (4, 16 and 64 μg/mL) and tigecycline (0.25, 1 and 4 μg/mL), at 12, 24 and 48 h. Afterwards, CFUs were determined plating on blood agar to measure the surviving intracellular bacteria. The viability of PCLS was assessed by Alamar Blue assay and corroborated using histopathological analysis. PCLS were successfully infected with a virulent strain of M. abscessus as demonstrated by CFUs and detailed histopathological analysis. The time-course infection, including tissue damage, parallels in vivo findings reported in genetically modified murine models for M. abscessus infection. Tigecycline showed a bactericidal effect at 48 h that achieved a reduction of > 4log10 CFU/mL against the intracellular mycobacteria, while imipenem showed a bacteriostatic effect. The use of this new organotypic ex vivo model provides the opportunity to test new drugs against M. abscessus, decreasing the use of costly and tedious animal models.

中文翻译:

脓肿分枝杆菌对鼠精确切割肺组织切片的离体感染:研究抗分枝杆菌药物的模型

由脓肿分枝杆菌引起的多重耐药感染通常需要复杂且长期的治疗方案。在这里,我们报告了使用小鼠精确切割肺切片的器官型培养物对新的离体抗菌药敏试验模型的评估,这是一种实验模型,其中代谢活动和器官的所有常见细胞类型都被发现,同时组织结构和保持不同细胞之间的相互作用。使用 Krumdieck® 组织切片机从野生型 BALB/c 小鼠的肺制备精确切割的肺切片 (PCLS)。肺组织切片是用有毒的脓肿分枝杆菌菌株 L948 离体感染的。然后,我们在 12、24 和 48 小时测试了两种药物的抗菌活性:亚胺培南(4、16 和 64 μg/mL)和替加环素(0.25、1 和 4 μg/mL)。然后,在血琼脂平板上测定 CFU,以测量存活的细胞内细菌。PCLS 的生存能力通过 Alamar Blue 测定进行评估,并使用组织病理学分析加以证实。正如 CFU 和详细的组织病理学分析所证明的那样,PCLS 成功地感染了 M. abscessus 的毒株。时程感染,包括组织损伤,与脓肿分枝杆菌感染的转基因小鼠模型中报道的体内发现相似。替加环素在 48 小时时显示出杀菌作用,对细胞内分枝杆菌的作用降低了 > 4log10 CFU/mL,而亚胺培南显示出抑菌作用。使用这种新的器官型离体模型提供了测试针对脓肿分枝杆菌的新药物的机会,减少了昂贵且繁琐的动物模型的使用。
更新日期:2020-11-22
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