Bednar Kyle J, Hardy Lakeya, Smeekens Johanna, Raghuwanshi Dharmendra, Duan Shiteng, Kulis Mike D, Macauley Matthew S
Janssen R&D.
Department of Microbiology and Immunology, University of North Carolina; UNC Food Allergy Initiative, University of North Carolina.
J Vis Exp. 2018 Oct 25(140):58285. doi: 10.3791/58285.
Antibody responses provide critical protective immunity to a wide array of pathogens. There remains a high interest in generating robust antibodies for vaccination as well as understand how pathogenic antibody responses develop in allergies and autoimmune disease. Generating robust antigen-specific antibody responses is not always trivial. In mouse models, it often requires multiple rounds of immunizations with adjuvant that leads to a great deal of variability in the levels of induced antibodies. One example is in mouse models of peanut allergies where more robust and reproducible models that minimize mouse numbers and the use of adjuvant would be beneficial. Presented here is a highly reproducible mouse model of peanut allergy anaphylaxis. This new model relies on two key factors: (1) antigen-specific splenocytes are adoptively transferred from a peanut-sensitized mouse into a naïve recipient mouse, normalizing the number of antigen-specific memory B- and T-cells across a large number of mice; and (2) recipient mice are subsequently boosted with a strong multivalent immunogen in the form of liposomal nanoparticles displaying the major peanut allergen (Ara h 2). The major advantage of this model is its reproducibility, which ultimately lowers the number of animals used in each study, while minimizing the number of animals receiving multiple injections of adjuvant. The modular assembly of these immunogenic liposomes provides relatively facile adaptability to other allergic or autoimmune models that involve pathogenic antibodies.
抗体反应为多种病原体提供关键的保护性免疫。人们对于研发用于疫苗接种的强效抗体以及了解致病性抗体反应在过敏和自身免疫性疾病中如何发展仍有着浓厚兴趣。产生强效的抗原特异性抗体反应并非总是轻而易举的。在小鼠模型中,通常需要用佐剂进行多轮免疫,这会导致诱导抗体水平出现很大差异。一个例子是在花生过敏的小鼠模型中,若能有更强大且可重复的模型,将小鼠数量和佐剂的使用降至最低,那将是有益的。本文介绍了一种高度可重复的花生过敏过敏反应小鼠模型。这个新模型依赖于两个关键因素:(1)将抗原特异性脾细胞从花生致敏小鼠过继转移到未致敏的受体小鼠体内,使大量小鼠体内抗原特异性记忆B细胞和T细胞的数量标准化;(2)随后用展示主要花生过敏原(Ara h 2)的脂质体纳米颗粒形式的强效多价免疫原对受体小鼠进行加强免疫。该模型的主要优点是其可重复性,这最终减少了每项研究中使用的动物数量,同时将接受多次佐剂注射的动物数量降至最低。这些免疫原性脂质体的模块化组装为涉及致病性抗体的其他过敏或自身免疫模型提供了相对简便的适应性。