Meyer Christine, Haberthur Kristen, Asquith Mark, Messaoudi Ilhem
Division of Pathobiology and Immunology, Oregon National Primate Research Center, Beaverton, OR, USA.
Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR, USA.
Methods Mol Biol. 2015;1343:65-80. doi: 10.1007/978-1-4939-2963-4_6.
Flow cytometry is an invaluable technique that can be used to phenotypically and functionally characterize immune cell populations ex vivo. This technology has greatly advanced our ability to gain critical insight into age-related changes in immune function, commonly known as immune senescence. Rodents have been traditionally used to investigate the molecular mechanisms of immune senescence because they offer the distinct advantages of an extensive set of reagents, the presence of genetically modified strains, and a short lifespan that allows for longevity studies of short duration. More recently, nonhuman primates (NHPs), and specifically rhesus macaques, have emerged as a leading translational model to study various aspects of human aging. In contrast to rodents, they share significant genetic homology as well as physiological and behavioral characteristics with humans. Furthermore, rhesus macaques are a long-lived outbred species, which makes them an ideal translational model. Therefore, NHPs offer a unique opportunity to carry out mechanistic studies under controlled laboratory conditions (e.g., photoperiod, temperature, diet, and medications) in a species that closely mimics human biology. Moreover similar techniques (e.g., activity recording and MRI) can be used to measure physiological parameters in NHPs, making direct comparisons between NHP and human data sets possible. In addition, the outbred genetics of NHPs enables rigorous validation of research findings that goes beyond proof of principle. Finally, self-selection bias that is often unavoidable in human clinical trials can be completely eliminated with NHP studies. Here we describe flow cytometry-based methods to phenotypically and functionally characterize innate immune cells as well as T and B lymphocyte subsets from isolated peripheral blood mononuclear cells (PBMC) in rhesus macaques.
流式细胞术是一项非常有价值的技术，可用于在体外对免疫细胞群体进行表型和功能特征分析。这项技术极大地提升了我们深入了解免疫功能中与年龄相关变化（即通常所说的免疫衰老）的能力。传统上，啮齿动物被用于研究免疫衰老的分子机制，因为它们具有一系列独特优势，如大量的试剂、存在基因修饰品系以及寿命较短，这使得进行短期的长寿研究成为可能。最近，非人类灵长类动物（NHPs），特别是恒河猴，已成为研究人类衰老各个方面的主要转化模型。与啮齿动物不同，它们与人类具有显著的基因同源性以及生理和行为特征。此外，恒河猴是一种长寿的远交物种，这使其成为理想的转化模型。因此，非人类灵长类动物提供了一个独特的机会，能够在受控的实验室条件下（如光照周期、温度、饮食和药物），在一个与人类生物学非常相似的物种中开展机制研究。此外，类似的技术（如活动记录和磁共振成像）可用于测量非人类灵长类动物的生理参数，从而使非人类灵长类动物和人类数据集之间的直接比较成为可能。此外，非人类灵长类动物的远交遗传学能够对研究结果进行严格验证，而不仅仅是原理验证。最后，在人类临床试验中常常不可避免的自我选择偏差，在非人类灵长类动物研究中可以完全消除。在此，我们描述基于流式细胞术的方法，用于对恒河猴分离的外周血单个核细胞（PBMC）中的固有免疫细胞以及T和B淋巴细胞亚群进行表型和功能特征分析。
