鞭虫感染muris：在肠道内的2型免疫和炎症模型
Summary
鞭muris感染肠道模型是一种Th2型免疫抗小鼠Th2反应产生保护和易感小鼠产生一种病态的Th1反应。
Abstract
鞭muris是一个自然的小鼠的病原体和鞭虫物种感染人类和牲畜 1的生物学和抗原性相似。感染性虫卵经口灌胃，在远端小肠孵化，侵入肠上皮细胞（IECS），该行盲肠和近端结肠隐窝和成熟后的蠕虫释放到环境中的鸡蛋1。这个模型是一个功能强大的工具来检查因素控制的CD4 +辅助性T细胞（TH）的激活，以及在肠上皮细胞的变化。免疫反应的抗近交系C57BL / 6和BALB / C，如发生的特点，是由Th2细胞极化的细胞因子（IL – 4，IL – 5和IL – 13）和驱逐的蠕虫病毒，而Th1相关的细胞因子（IL -12，IL – 18，IFN -γ）促进在遗传易感性的AKR / J 小鼠 2-6的慢性感染。 Th2细胞因子，促进生理肠道微环境的变化，包括IECS的快速周转，杯状细胞的分化，招聘和上皮通透性和平滑肌收缩，所有这一切都已经在蠕虫驱逐7-15牵连的变化。在这里，我们详细为传播鞭muris可以在随后的实验中使用的鸡蛋的的协议。我们还提供了为感染后的分析建议的样品实验收获。总体而言，该协议将提供的基本工具的研究人员，执行鞭muris小鼠感染模型，它可以用来解决有关的问题在胃肠道以及免疫效应功能的IECS钍倾向。
Protocol
1。传播鞭muris蛋要生成新的批次的鞭muris蛋，感染20-30免疫缺陷小鼠（如NOD.Cg Prkdc SCID IL2RG tm1Wjl / SZJ（NSG）或129S6/SvEvTac-Rag2 tm1Fwa（RAG2 – / – ））或遗传易感性小鼠（如AKR / J）6-8周约300鞭灌胃muris鸡蛋。 CO 2窒息牺牲后32-35天的小鼠。 揭露鼠标的腹侧方和湿用70％乙醇的腹部。 使用镊子把握腹部皮肤，使一个小切口，?…
Discussion
该协议的细节了一个标准的高剂量急性鞭muris感染研究者的要求，可以修改。例如，老鼠可以牺牲和组织在不同的日子里收获。要确定小鼠已成功建立了完整的蠕虫负担，他们可以被牺牲的第14天，在这一点所有小鼠，应随身携带了约200蠕虫的负担。也可被感染小鼠的32天检测到任何蠕虫将达到成熟阶段，并会维持其生命期限的主机。此外，该协议可以适应慢性鞭虫muris感染模型。要?…
Divulgaciones
The authors have nothing to disclose.
Acknowledgements
这项工作是支持的加拿大卫生研究院（MSH – 95368，澳门币89773和MOP – 106623，以锆石）和加拿大创新基金会。 SCM是一个胃肠病学博士后CIHR /加拿大协会的收件人。锆石是一种新CIHR调查。
Materials
| Name of the reagent | Company | Catalogue number | Comments |
|---|---|---|---|
| Animal Feeding Needles (18 x 1½”) | Popper | 7912 | |
| Smooth curved Forceps | Roboz | RS-5047 | |
| DMEM | Gibco | 11965 | |
| NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) | Jackson Laboratories | 005557 | These are the mice we used, however, any immunodeficient mice or susceptible strain should work. |
| RNAlater | Qiagen | 76104 | |
| 2 ml tubes | Axygen | MCT-200-C | |
| 15 ml tubes | Falcon | 352096 | |
| 6 well plates | Falcon | 353046 | |
| Paraformaldehyde | Electron Microscopy Science | 15710 | |
| α-RELMβ antibody | PeproTech Inc | 0694270Rb |
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Tags
Trichuris Muris InfectionType 2 ImmunityInflammation In The GutIntestinal Epithelial CellsCD4+ T Helper Cell ActivationResistant Inbred StrainsTh2 Polarized CytokinesTh1-associated CytokinesIntestinal Microenvironment ChangesWorm ExpulsionTrichuris Muris EggsExperimental HarvestPost-infection Analysis
Citar este artículo
Antignano, F., Mullaly, S. C., Burrows, K., Zaph, C. Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut. J. Vis. Exp. (51), e2774, doi:10.3791/2774 (2011).