Chattopadhyay Avik, Joseph Joel P, Shyam Sai, Nandi Dipankar
Department of Biochemistry, Indian Institute of Science.
Center for BioSystems Science and Engineering, Indian Institute of Science.
J Vis Exp. 2022 Jul 29(185). doi: 10.3791/63695.
Sepsis is a dysregulated host immune response to microbial invasion or tissue damage, leading to organ injury at a site distant from that of the infection or damage. Currently, the widely used mice models of sepsis include lipopolysaccharide (LPS)-induced endotoxemia, cecal ligation and puncture (CLP), and monobacterial infection model systems. This protocol describes a method to study the host responses during Salmonella Typhimurium infection-induced septic peritonitis in mice. S. Typhimurium, a Gram-negative intracellular pathogen, causes typhoid-like disease in mice. This protocol elaborates the culture preparation, induction of septic peritonitis in mice through intraperitoneal injection, and methods to study systemic host responses. Furthermore, the assessment of bacterial burden in different organs and the flow cytometric analysis of increased neutrophil numbers in the peritoneal lavage is presented. Salmonella Typhimurium-induced sepsis in mice leads to an increase in proinflammatory cytokines and rapid infiltration of neutrophils in the peritoneal cavity, leading to lower survival. Every step in this protocol has been optimized, resulting in high reproducibility of the pathogenesis of septic peritonitis. This model is useful for studying immunological responses during bacterial sepsis, the roles of different genes in disease progression, and the effects of drugs to attenuate sepsis.
脓毒症是宿主对微生物入侵或组织损伤的免疫反应失调,导致在远离感染或损伤部位的器官发生损伤。目前,广泛使用的脓毒症小鼠模型包括脂多糖(LPS)诱导的内毒素血症、盲肠结扎和穿刺(CLP)以及单菌感染模型系统。本方案描述了一种研究小鼠鼠伤寒沙门氏菌感染诱导的脓毒症性腹膜炎期间宿主反应的方法。鼠伤寒沙门氏菌是一种革兰氏阴性细胞内病原体,可在小鼠中引起伤寒样疾病。本方案阐述了培养物制备、通过腹腔注射诱导小鼠脓毒症性腹膜炎以及研究全身宿主反应的方法。此外,还介绍了不同器官中细菌载量的评估以及腹腔灌洗中嗜中性粒细胞数量增加的流式细胞术分析。鼠伤寒沙门氏菌诱导的小鼠脓毒症会导致促炎细胞因子增加以及嗜中性粒细胞在腹腔内迅速浸润,从而导致存活率降低。本方案中的每一步都经过了优化,使得脓毒症性腹膜炎发病机制具有高度可重复性。该模型可用于研究细菌性脓毒症期间的免疫反应、不同基因在疾病进展中的作用以及药物减轻脓毒症的效果。