Division of Host-Microbe Systems & Therapeutics, Department of Pediatrics, University of California San Diego, La Jolla, California.
School of Life Sciences, Freising-Weihenstephan, ZIEL - Institute for Food & Health, Freising-Weihenstephan, Technical University of Munich, Germany.
Curr Protoc. 2023 Jul;3(7):e824. doi: 10.1002/cpz1.824.
The pathogen Salmonella enterica encompasses a range of bacterial serovars that cause intestinal inflammation and systemic infections in humans. Mice are a widely used infection model due to their relative simplicity and versatility. Here, we provide standardized protocols for culturing the prolific zoonotic pathogen S. enterica serovar Typhimurium for intragastric inoculation of mice to model colitis or systemic dissemination, along with techniques for direct extraintestinal infection. Furthermore, we present procedures for quantifying pathogen burden and for characterizing the immune response by analyzing tissue pathology, inflammatory markers, and immune cells from intestinal tissues. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Murine colitis model utilizing oral streptomycin pretreatment and oral S. Typhimurium administration Basic Protocol 2: Intraperitoneal injection of S. Typhimurium for modeling extraintestinal infection Support Protocol 1: Preparation of S. Typhimurium inoculum Support Protocol 2: Preparation of mixed S. Typhimurium inoculum for competitive infection Basic Protocol 3: Assessment of S. Typhimurium burden Support Protocol 3: Preservation and pathological assessment of S. Typhimurium-infected tissues Support Protocol 4: Measurement of inflammatory marker expression in intestinal tissues by qPCR Support Protocol 5: Preparation of intestinal content for inflammatory marker quantification by ELISA Support Protocol 6: Immune cell isolation from Salmonella-infected intestinal tissues.
病原体沙门氏菌包括一系列细菌血清型，可引起人类肠道炎症和全身感染。由于其相对简单和多功能，小鼠是广泛使用的感染模型。在这里，我们提供了标准化的方案，用于培养丰富的人畜共患病原体鼠伤寒沙门氏菌血清型，用于通过胃内接种感染小鼠以模拟结肠炎或全身传播，以及直接肠外感染的技术。此外，我们还介绍了用于量化病原体负担和通过分析肠道组织的组织病理学、炎症标志物和免疫细胞来描述免疫反应的程序。© 2023 作者。Wiley Periodicals LLC 出版的现行方案。基本方案 1：利用口服链霉素预处理和口服鼠伤寒沙门氏菌给药的小鼠结肠炎模型基本方案 2：腹腔注射鼠伤寒沙门氏菌以模拟肠外感染支持方案 1：鼠伤寒沙门氏菌接种物的制备支持方案 2：用于竞争感染的混合鼠伤寒沙门氏菌接种物的制备基本方案 3：鼠伤寒沙门氏菌负荷的评估支持方案 3：保存和病理评估感染鼠伤寒沙门氏菌的组织支持方案 4：通过 qPCR 测量肠道组织中炎症标志物的表达支持方案 5：通过 ELISA 量化肠道内容物中炎症标志物的含量支持方案 6：从沙门氏菌感染的肠道组织中分离免疫细胞。
