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在大鼠腹膜透析膜室内培养问号钩端螺旋体。

Cultivation of Leptospira interrogans Within Rat Peritoneal Dialysis Membrane Chambers.

机构信息

Department of Medicine, University of Connecticut Health, Farmington, CT, USA.

Departments of Medicine, Pediatrics, and Molecular Biology and Biophysics, University of Connecticut Health, Farmington, CT, USA.

出版信息

Methods Mol Biol. 2020;2134:229-242. doi: 10.1007/978-1-0716-0459-5_21.

Abstract

In order to sustain its zoonotic lifecycle, leptospires must adapt to growth within the host milieu. Signals encountered within the mammal also trigger regulatory programs required by Leptospira for the expression of virulence-related gene products. The complex transcriptional, antigenic, and physiological changes leptospires undergo within the mammal are collectively referred to as "host adaptation." In this chapter, we describe the procedures for the generation of host-adapted Leptospira spp. by cultivation within dialysis membrane chambers (DMCs) implanted in rat peritoneal cavities. In this model, Leptospira spp. diluted in EMJH medium are sequestered within sterile dialysis membrane tubing closed at both ends. The chamber then is surgically implanted within the peritoneal cavity of a rat and incubated for 7-10 days. During this period, leptospires are exposed to many, if not all, of the physiological and nutritional cues required for host adaptation while at the same time protected from clearance by host innate and adaptive immune defenses.

摘要

为了维持其人畜共患的生命周期,钩端螺旋体必须适应在宿主环境中生长。哺乳动物体内遇到的信号也会触发钩端螺旋体表达毒力相关基因产物所需的调节程序。钩端螺旋体在哺乳动物体内经历的复杂转录、抗原和生理变化被统称为“宿主适应”。在本章中,我们描述了通过在植入大鼠腹腔的透析膜室(DMC)中培养来生成适应宿主的钩端螺旋体 spp. 的程序。在该模型中,在 EMJH 培养基中稀释的钩端螺旋体 spp. 被封闭在两端的无菌透析膜管中隔离。然后,该室通过手术植入大鼠腹腔内,并孵育 7-10 天。在此期间,钩端螺旋体暴露于许多(如果不是全部)适应宿主所需的生理和营养线索,同时受到宿主先天和适应性免疫防御的保护而不被清除。

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