OMIP-075
A 22-color panel for the measurement of antigen-specific T-cell responses in human and nonhuman primates
Cytometry PART A, Volume 99, Issue 9, 884-887 (2021)
Takuto Nogimori, Eiko Moriishi, Mami Ikeda, Shokichi Takahama, Takuya Yamamoto
Description: This panel was designed to assess the antigen-specific CD4+ T-cell responses in human and non-human primates in order to monitor immune response kinetics in infectious diseases and to evaluate efficacy and safety in the development of vaccines against COVID-19. This panel separates CD4+ T cells into Th1, Th2, Th17, and peripheral Tfh (pTfh) by measuring the expression of chemokine receptors (CXCR3, CCR4, CCR6,and CXCR5). Furthermore, it includes antibodies that detect an activation marker (CD154), a degranulation marker (CD107a), cytokines (IFNγ,TNF, IL-2, IL-4, IL-13, IL-17, and IL-21), a chemokine (MIP-1β/CCL4), and immune checkpoint/activation molecules (PD-1 and ICOS). We have optimized this panel for cryopreserved human and cynomolgus macaque peripheral blood mononuclear cells (PBMCs) with antigen-specific stimulation. This panel will be useful not only for finding immunological correlates of protection against infectious diseases and/or cancer but also for evaluating immune responses after vaccination.
Species: Human, Cynomolgus macaque
Tissue: Peripheral blood
Cell Subset: T cell, Ctl, T helper, T memory, Tcrgd, Th1, Th17, Th2, Tfh
Keyword: Antigen-specific t cell response, Cryopreserved pbmc, Immune checkpoint, Vaccination
Machine: FACSymphony A5
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| Live/Dead Fix Blue | All Species | Live/Dead Fix Blue | Viability | |||
| CD27 | Human | 1A4CD27 | PE-Cy5 | Differentiation for human | Hu mem, Th1, Th2, Th17, Tfh | Med |
| CD28 | Human | CD28.2 | Brilliant Ultraviolet 805 | Differentiation for monkey | Macaque mem | Med |
| PD-1 | Human | EH12.2H7 | Brilliant Violet 750 | Immune checkpoint | T helper | Low |
| CD278 | Human | C398.4A | Brilliant Ultraviolet 395 | Immune checkpoint | T helper | Low |
| CD183 | Human | G025H7 | PE | Th1 marker | Th1 | Med |
| CD185 | Human | MU5UBEE | PE-eFluor 610 | pTfh marker | Tfh | Low |
| CD194 | Human | L291H4 | Brilliant Violet 510 | Th2 marker | Th2 | Med |
| CCR6 | Human | G034E3 | APC-Fire 750 | Th17 marker | Th17 | Low |
| CD40L | Human | TRAP1 | FITC | Activation marker | Hu mem, Macaque mem, Th1, Th2, Th17, Tfh | Med |
| IL-4 | Human | 8D4-8 | PE-Cy7 | Cytokine | Th2, Tfh | Low |
| IL-13 | Human | JES10-5A2 | Brilliant Violet 421 | Cytokine | Tfh | Low |
| IL-17A | Human | BL168 | Brilliant Violet 605 | Cytokine | Th17 | Low |
| IL-21 | Human | 3A3-N2.1 (also known as 3A3-N2) | Alexa Fluor 647 | Cytokine | Tfh | Low |
| MIP-1 beta | Human | D21-1351 | Alexa Fluor 700 | Inflammation | T helper, CTL | Med |
| IFN-gamma | Human | 4S.B3 | Brilliant Violet 786 | Cytokine | T helper, CTL | Med |
| CD107a | Human | H4A3 | Brilliant Violet 711 | Degranulation | CTL | Med |
| CD95 | Human | Dx2 | PE-Cy5 | Differentiation for monkey | Tmem | Low |
| CD8a | Human | RPA-T8 | Brilliant Ultraviolet 563 | Lineage | CTL, Macaque mem, Hu mem | High |
| TNF | Human | Mab11 | Brilliant Violet 650 | Cytokine | T helper, CTL | Med |
| IL-2 | Human | MQ1-17H12 | Brilliant Ultraviolet 737 | Intracellular Staining | Th1, Tfh | Med |
| CD4 | Human | S3.5 | PE-Cy5.5 | Lineage | Th1, Th2, Th17, Tfh, Tmem | High |
| CD3 | Human | SP34-2 | Brilliant Ultraviolet 615 | Lineage | T helper, CTL, Th1, Th2, Th17, Tfh, Macaque mem, Hu mem | High |
| CD45RO | Human | UCHL1 | Brilliant Ultraviolet 805 | Differentiation for human | Hu mem, T helper, CTL, Th1, Th2, Th17, Tfh | Med |
