tetano
Editor, Senior Moderator
Immunol Cell Biol
. 2020 Aug 1.
doi: 10.1111/imcb.12383. Online ahead of print.
Immune profiling of influenza-specific B cell and T cell responses in macaques using flow cytometry-based assays
Marios Koutsakos[SUP] 1 [/SUP], Toshiki Sekiya[SUP] 1 2 3 [/SUP], Brendon Y Chua[SUP] 1 2 3 [/SUP], Thi O H Nguyen[SUP] 1 [/SUP], Adam K Wheatley[SUP] 1 [/SUP], Jennifer A Juno[SUP] 1 [/SUP], Marumi Ohno[SUP] 2 [/SUP], Naoki Nomura[SUP] 2 [/SUP], Yuki Ohara[SUP] 4 [/SUP], Tomohiro Nishimura[SUP] 4 [/SUP], Masafumi Endo[SUP] 4 [/SUP], Saori Suzuki[SUP] 5 [/SUP], Hirohito Ishigaki[SUP] 5 [/SUP], Misako Nakayama[SUP] 5 [/SUP], Cong Thanh Nguyen[SUP] 5 [/SUP], Yasushi Itoh[SUP] 5 [/SUP], Masashi Shingai[SUP] 2 3 [/SUP], Kazumasa Ogasawara[SUP] 5 [/SUP], Yoichiro Kino[SUP] 6 [/SUP], Stephen J Kent[SUP] 1 7 8 [/SUP], David C Jackson[SUP] 1 2 3 [/SUP], Lorena E Brown[SUP] 1 2 3 [/SUP], Hiroshi Kida[SUP] 2 3 9 [/SUP], Katherine Kedzierska[SUP] 1 [/SUP]
Affiliations
Abstract
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[SUP]+[/SUP] and CD4[SUP]+[/SUP] 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 cells 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 towards influenza viruses can be also adapted to understanding immunity to other infections or vaccination approaches in macaque models.
Keywords: flow cytometry; inflammation; influenza-specific B cells; influenza-specific T cells; macaques.
. 2020 Aug 1.
doi: 10.1111/imcb.12383. Online ahead of print.
Immune profiling of influenza-specific B cell and T cell responses in macaques using flow cytometry-based assays
Marios Koutsakos[SUP] 1 [/SUP], Toshiki Sekiya[SUP] 1 2 3 [/SUP], Brendon Y Chua[SUP] 1 2 3 [/SUP], Thi O H Nguyen[SUP] 1 [/SUP], Adam K Wheatley[SUP] 1 [/SUP], Jennifer A Juno[SUP] 1 [/SUP], Marumi Ohno[SUP] 2 [/SUP], Naoki Nomura[SUP] 2 [/SUP], Yuki Ohara[SUP] 4 [/SUP], Tomohiro Nishimura[SUP] 4 [/SUP], Masafumi Endo[SUP] 4 [/SUP], Saori Suzuki[SUP] 5 [/SUP], Hirohito Ishigaki[SUP] 5 [/SUP], Misako Nakayama[SUP] 5 [/SUP], Cong Thanh Nguyen[SUP] 5 [/SUP], Yasushi Itoh[SUP] 5 [/SUP], Masashi Shingai[SUP] 2 3 [/SUP], Kazumasa Ogasawara[SUP] 5 [/SUP], Yoichiro Kino[SUP] 6 [/SUP], Stephen J Kent[SUP] 1 7 8 [/SUP], David C Jackson[SUP] 1 2 3 [/SUP], Lorena E Brown[SUP] 1 2 3 [/SUP], Hiroshi Kida[SUP] 2 3 9 [/SUP], Katherine Kedzierska[SUP] 1 [/SUP]
Affiliations
- PMID: 32741011
- DOI: 10.1111/imcb.12383
Abstract
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[SUP]+[/SUP] and CD4[SUP]+[/SUP] 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 cells 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 towards influenza viruses can be also adapted to understanding immunity to other infections or vaccination approaches in macaque models.
Keywords: flow cytometry; inflammation; influenza-specific B cells; influenza-specific T cells; macaques.