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Immune profiling of influenza-specific B- and T-cell responses in macaques using flow cytometry-based assays.
Immunology and Cell Biology ( IF 4 ) Pub Date : 2020-08-01 , DOI: 10.1111/imcb.12383
Marios Koutsakos 1 , Toshiki Sekiya 1, 2, 3 , Brendon Y Chua 1, 2, 3 , Thi Hoang Oanh Nguyen 1 , Adam K Wheatley 1 , Jennifer A Juno 1 , Marumi Ohno 2 , Naoki Nomura 2 , Yuki Ohara 4 , Tomohiro Nishimura 4 , Masafumi Endo 4 , Saori Suzuki 5 , Hirohito Ishigaki 5 , Misako Nakayama 5 , Cong T Nguyen 5 , Yasushi Itoh 5 , Masashi Shingai 2, 3 , Kazumasa Ogasawara 5 , Yoichiro Kino 6 , Stephen J Kent 1, 7, 8 , David C Jackson 1, 2, 3 , Lorena E Brown 1, 2, 3 , Hiroshi Kida 2, 3, 9 , Katherine Kedzierska 1, 3
Affiliation  

Influenza remains a significant global public health burden, despite substantial annual vaccination efforts against circulating virus strains. As a result, novel vaccine approaches are needed to generate long‐lasting and universal broadly cross‐reactive immunity against distinct influenza virus strains and subtypes. Several new vaccine candidates are currently under development and/or in clinical trials. The successful development of new vaccines requires testing in animal models, other than mice, which capture the complexity of the human immune system. Importantly, following vaccination or challenge, the assessment of adaptive immunity at the antigen‐specific level is particularly informative. In this study, using peripheral blood mononuclear cells (PBMCs) from cynomolgus macaques, we describe detection methods and in‐depth analyses of influenza virus‐specific B cells by recombinant hemagglutinin probes and flow cytometry, as well as the detection of influenza virus‐specific CD8+ and CD4+ T cells by stimulation with live influenza A virus and intracellular cytokine staining. We highlight the potential of these assays to be used with PBMCs from other macaque species, including rhesus macaques, pigtail macaques and African green monkeys. We also demonstrate the use of a human cytometric bead array kit in detecting inflammatory cytokines and chemokines from cynomolgus macaques to assess cytokine/chemokine milieu. Overall, the detection of influenza virus‐specific B and T cells, together with inflammatory responses, as described in our study, provides useful insights for evaluating novel influenza vaccines. Our data deciphering immune responses toward influenza viruses can be also adapted to understanding immunity to other infections or vaccination approaches in macaque models.

中文翻译:

使用基于流式细胞术的检测对猕猴流感特异性 B 和 T 细胞反应的免疫分析。

尽管每年都针对流行的病毒株进行了大量的疫苗接种工作,但流感仍然是一个重大的全球公共卫生负担。因此,需要新的疫苗方法来产生针对不同流感病毒株和亚型的持久且普遍的广泛交叉反应免疫。几种新的候选疫苗目前正在开发和/或临床试验中。新疫苗的成功开发需要在小鼠以外的动物模型中进行测试,这些模型捕捉了人类免疫系统的复杂性。重要的是,在接种疫苗或激发后,抗原特异性水平的适应性免疫评估特别有用。在这项研究中,使用食蟹猴的外周血单核细胞 (PBMC),+和 CD4 +通过用活甲型流感病毒和细胞内细胞因子染色刺激 T 细胞。我们强调了这些检测与来自其他猕猴物种的 PBMC 一起使用的潜力,包括恒河猴、猪尾猕猴和非洲绿猴。我们还展示了使用人细胞计数珠阵列试剂盒检测食蟹猴的炎性细胞因子和趋化因子,以评估细胞因子/趋化因子环境。总体而言,如我们的研究中所述,流感病毒特异性 B 和 T 细胞的检测以及炎症反应为评估新型流感疫苗提供了有用的见解。我们对流感病毒免疫反应的破译数据也可以用于理解猕猴模型中对其他感染或疫苗接种方法的免疫力。
更新日期:2020-08-01
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