Ethanol-fed, CpG-treated mice: Listeria susceptibility

乙醇喂养、CpG 处理的小鼠：李斯特菌易感性

• 批准号: 6478290
• 负责人: NANCY B RAY
• 金额: $ 11.79万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6478290
• 关键词: CpG islands; DNA; Listeria; Listeria infections; Peyer's patches; alcoholic beverage consumption; chemoprevention; cytokine; disease /disorder proneness /risk; enzyme linked immunosorbent assay; ethanol; gene expression; gene targeting; genetically modified animals; helper T lymphocyte; host organism interaction; immunity; immunomagnetic separation; immunosuppression; immunotherapy; laboratory mouse; leukocyte activation /transformation; macrophage; mucosal immunity; nonhuman therapy evaluation; tissue /cell culture

DESCRIPTION (provided by applicant): Chronic ethanol consumption causes immunosuppression, particularly related to Th1 cytokine expression, and causes increased susceptibility to infection including infection with the intracellular bacterial human pathogen, Listeria monocytogenes. CpG DNA induces innate Thl-like responses and reduced susceptibility to Listeria monocytogenes detected by decreased replication of L. monocytogenes in murine spleens and livers. CpG immunostimulatory DNA will be used to stimulate innate systemic and mucosal immune responses in ethanol- fed and water-fed control mice. Induction of cytokine expression in vitro and in vivo in spleen and Peyer's patch cells from CpG-treated, ethanol-fed and water-fed control mice will be detected by enzyme-linked immunosorbant assay (ELISA), and specific Thl cytokine expressing cells will be identified by magnetic cell sorting (MACS) and intracellular cytokine staining. CpG DNA treatment and immunostimulation will be used to overcome ethanol-induced immunosuppression, which will be detected by reduced replication of L. monocytogenes in spleens and livers by a colony forming (CFU) assay. Various routes and doses of CpG treatment and challenge with L. monocytogenes including intraperitoneal (IP), intravenous (IV), and oral will be used to elicit both systemic and mucosal immunity and to determine the limits of protection induced by CpG DNA. Reduced replication of L. monocytogenes will be correlated with cytokine expression detected by ELISA and the mechanism of CpG-induced reduction in replication will be determined by identifying by flow cytometry and intracellular cytokine staining whether specific cells may be suppressed or depleted by chronic ethanol consumption and stimulated by CpG treatment. Because defects in killing Mycobacterium avium, an intracellular bacteria similar to L. monocytogenes and to the organism that causes tuberculosis, have been demonstrated for macrophages following exposure to alcohol, intracellular killing of L. monocytogenes by peritoneal macrophages will be examined to determine whether ethanol-fed mice are deficient in killing L. monocytogenes and whether CpG DNA stimulation may enhance killing. These studies could lead to treatments for protecting alcoholic patients from intracellular bacterial infections such as those caused by L. monocytogenes or Mycobacterium tuberculosis (TB).

描述（由申请人提供）：慢性乙醇消耗原因 免疫抑制，尤其与Th1细胞因子表达有关，并导致 增加感染的敏感性，包括感染 细胞内细菌人病原体，单核细胞增生李斯特菌。 CpG DNA 引起先天THL样的反应并降低对李斯特菌的敏感性 通过鼠中单核细胞增生的复制降低检测到的单核细胞增生 脾脏和肝脏。 CPG免疫刺激性DNA将用于刺激先天 乙醇和水馈控制中的全身和粘膜免疫反应 老鼠。在体外和体内诱导细胞因子表达在脾和体内 PEYER的斑块细胞来自CpG处理的，乙醇喂养的和水喂的对照小鼠 将通过酶联免疫吸附测定（ELISA）检测到 THL细胞因子表达细胞将通过磁细胞分类鉴定 （MAC）和细胞内细胞因子染色。 CpG DNA处理和 免疫刺激将用于克服乙醇诱导的免疫抑制， 通过减少脾中的单核细胞增生李斯特菌的复制来检测到这一点 和肝脏形成（CFU）测定法。各种路线和剂量CpG 腹膜内（IP）的单核细胞增生李斯特菌的治疗和挑战， 静脉注射（IV）和口服将用于引起全身和粘膜 免疫力并确定CpG DNA引起的保护极限。减少 单核细胞增生乳杆菌的复制将与细胞因子表达相关 通过ELISA检测到CPG诱导的复制减少的机制 通过通过流式细胞术和细胞内细胞因子来确定 染色是否可以被慢性抑制或耗尽特定细胞 乙醇消耗并通过CPG治疗刺激。因为缺陷 杀死类似于L的细胞内细菌杀死分枝杆菌。 单核细胞增生并致导致结核病的生物，已经是 暴露于酒精后，可用于巨噬细胞 将检查腹膜巨噬细胞杀死单核细胞增生李斯特氏菌。 确定乙醇喂养的小鼠是否缺乏杀死单核细胞增生乳杆菌 CpG DNA刺激是否可以增强杀伤。这些研究可能会导致 用于保护酒精性患者免受细胞内细菌的治疗 诸如单核细胞增生乳杆菌或分枝杆菌引起的感染诸如感染 结核病（TB）。