1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Department of Medicine, McGill University, Montreal, Quebec, Canada. [3] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Nat Protoc. 2014 Dec;9(12):2740-54. doi: 10.1038/nprot.2014.184. Epub 2014 Nov 6.
Heligmosomoides polygyrus bakeri (Hpb) infection in mice is a convenient model for studying the pathophysiology and immunology of gastrointestinal (GI) helminth infection. Hpb infection suppresses immune responses to bystander antigens and unrelated pathogens, and it slows the progression and modifies the outcome of immune-mediated diseases. Hpb-derived excretory-secretory (ES) products potently modulate CD4(+) helper T cell (TH) responses by inducing regulatory T cells, tolerogenic dendritic cells (DCs) and immunoregulatory cytokines. This observation has spiked interest in identifying the immunomodulatory molecules, especially proteins, in ES products from Hpb and other GI nematodes for development as novel therapies to treat individuals with immune-mediated diseases, such as inflammatory bowel diseases (IBDs). In this protocol, we describe how to (i) maintain Hpb in the laboratory for experimental infections, (ii) collect adult worms from infected mice to generate ES products and (iii) evaluate the modulatory effects of ES products on toll-like receptor (TLR) ligand-induced maturation of CD11c(+) DCs. The three major sections of the PROCEDURE can be used independently, and they require ∼6, 10 and 27 h, respectively. Although other methods use a modified Baermann apparatus to collect Hpb adult worms, we describe a method that involves dissection of adult worms from intestinal tissue. The protocol will be useful to investigators studying the host-parasite interface and identifying and analyzing helminth-derived molecules with therapeutic potential.
贝氏多房棘隙吸虫（Hpb）感染小鼠是研究胃肠道（GI）寄生虫感染的病理生理学和免疫学的一种方便模型。Hpb 感染抑制了对旁观者抗原和无关病原体的免疫反应，并且它减缓了免疫介导性疾病的进展并改变了其结果。Hpb 衍生的排泄-分泌（ES）产物通过诱导调节性 T 细胞、耐受性树突状细胞（DC）和免疫调节细胞因子，强烈调节 CD4（+）辅助 T 细胞（TH）反应。这一观察结果激发了人们对鉴定 ES 产物中免疫调节分子（尤其是蛋白质）的兴趣，这些分子来自 Hpb 和其他 GI 线虫，以开发治疗免疫介导性疾病（如炎症性肠病（IBD））患者的新型疗法。在本方案中，我们描述了如何（i）在实验室中维持 Hpb 以进行实验感染，（ii）从感染小鼠中收集成虫以产生 ES 产物，以及（iii）评估 ES 产物对 TLR 配体诱导的 CD11c（+）DC 成熟的调节作用。该 PROCEDURE 的三个主要部分可以独立使用，分别需要约 6、10 和 27 小时。尽管其他方法使用改良的贝氏装置收集 Hpb 成虫，但我们描述了一种涉及从肠组织中分离成虫的方法。该方案将有助于研究宿主-寄生虫界面的研究人员，并鉴定和分析具有治疗潜力的寄生虫衍生分子。
