Willis Alexandra R, Tamim El Jarkass Hala, Reinke Aaron W
Department of Molecular Genetics, University of Toronto.
Department of Molecular Genetics, University of Toronto;
J Vis Exp. 2022 Apr 6(182). doi: 10.3791/63636.
Inherited immunity describes how some animals can pass on the "memory" of a previous infection to their offspring. This can boost pathogen resistance in their progeny and promote survival. While inherited immunity has been reported in many invertebrates, the mechanisms underlying this epigenetic phenomenon are largely unknown. The infection of Caenorhabditis elegans by the natural microsporidian pathogen Nematocida parisii results in the worms producing offspring that are robustly resistant to microsporidia. The present protocol describes the study of intergenerational immunity in the simple and genetically tractable N. parisii -C. elegans infection model. The current article describes methods for infecting C. elegans and generating immune-primed offspring. Methods are also given for assaying resistance to microsporidia infection by staining for microsporidia and visualizing infection by microscopy. In particular, inherited immunity prevents host cell invasion by microsporidia, and fluorescence in situ hybridization (FISH) can be used to quantify invasion events. The relative amount of microsporidia spores produced in the immune-primed offspring can be quantified by staining the spores with a chitin-binding dye. To date, these methods have shed light on the kinetics and pathogen specificity of inherited immunity, as well as the molecular mechanisms underlying it. These techniques, alongside the extensive tools available for C. elegans research, will enable important discoveries in the field of inherited immunity.
遗传免疫描述了一些动物如何将先前感染的“记忆”传递给它们的后代。这可以增强其后代对病原体的抵抗力并促进生存。虽然在许多无脊椎动物中都报道了遗传免疫，但这种表观遗传现象背后的机制在很大程度上尚不清楚。天然微孢子虫病原体巴黎嗜线虫感染秀丽隐杆线虫会导致线虫产生对微孢子虫具有强大抵抗力的后代。本方案描述了在简单且易于遗传操作的巴黎嗜线虫-秀丽隐杆线虫感染模型中对代际免疫的研究。本文描述了感染秀丽隐杆线虫并产生免疫致敏后代的方法。还给出了通过对微孢子虫进行染色并通过显微镜观察感染情况来检测对微孢子虫感染的抵抗力的方法。特别地，遗传免疫可防止微孢子虫侵入宿主细胞，荧光原位杂交（FISH）可用于量化侵入事件。通过用几丁质结合染料对孢子进行染色，可以量化免疫致敏后代中产生的微孢子虫孢子的相对数量。迄今为止，这些方法已经揭示了遗传免疫的动力学和病原体特异性以及其背后的分子机制。这些技术，连同可用于秀丽隐杆线虫研究的大量工具，将推动遗传免疫领域的重要发现。
