158 peripheral immune cell subsets analyzed by flow cytometry
| 1. Total CD4+ T cells | 2. Total CD8+ T cells | 4. Total B cells |
| PD-L1+ CD4 – activation/inhibition | PD-L1+ CD8 – activation/inhibition | PD-L1+ B cells – activation/inhibition |
| PD-1+ CD4 – activation/inhibition | PD-1+ CD8 – activation/inhibition | PD-1+ B cells- activation/inhibition |
| CTLA-4+ CD4 – inhibition | CTLA-4+ CD8 – inhibition | 5. Total NK |
| Tim-3+ CD4 – inhibition | Tim-3+ CD8 – inhibition | PD-L1+ NK – inhibition |
| 41bb+ CD4 – costimulation | 41bb+ CD8 – costimulation | PD-1+ NK – activation/inhibition |
| Ki67+ CD4 – proliferation | Ki67+ CD8 – proliferation | Tim-3+ NK – activation/inhibition |
| CD73+ CD4 – exhausted/suppressive | CD73+ CD8 – exhausted/suppressive | Ki67+ NK – proliferation |
| ICOS+ CD4 – activation | Naïve (CCR7+CD45RA+) CD8 | NKp30+ NK – activation |
| ICOS+ PD-L1+ CD4 – activation/inhibition | Central memory (CCR7+CD45RA-) CD8 | NKp46+ NK – activation |
| ICOS+ PD-1+ CD4 – activation/inhibition | PD-L1+ CM CD8 – activation/inhibition | NKG2D+ NK – activation |
| Naïve (CCR7+CD45RA+) CD4 | PD-1+ CM CD8 – activation/inhibition | CD226+ NK – adhesion/activation |
| Central memory (CCR7+ CD45RA-) CD4 | CTLA-4+ CM CD8 – inhibition | Mature (CD16+ CD56dim) NK – lytic |
| PD-L1+ CM CD4 – activation/inhibition | Tim-3+ CM CD8 – inhibition | PD-L1+ mature NK – inhibition |
| PD-1+ CM CD4 – activation/inhibition | 41bb+ CM CD8 – costimulation | PD-1+ mature NK – activation/inhibition |
| CTLA-4+ CM CD4 – inhibition | Ki67+ CM CD8 – proliferation | Tim-3+ mature NK – activation/inhibition |
| Tim-3+ CM CD4 – inhibition | CD73+ CM CD8 – exhausted/suppressive | Ki67+ mature NK – proliferation |
| 41bb+ CM CD4 – costimulation | Effector memory (CCR7- CD45RA-) CD8 | NKp30+ mature NK – activation |
| Ki67+ CM CD4 – proliferation | PD-L1+ EM CD8 – activation/inhibition | NKp46+ mature NK – activation |
| CD73+ CM CD4 – exhausted/suppressive | PD-1+ EM CD8 - activation/inhibition | NKG2D+ mature NK – activation |
| Effector memory (CCR7- CD45RA-) CD4 | CTLA-4+ EM CD8 – inhibition | CD226+ mature NK – adhesion/activation |
| PD-L1+ EM CD4 – activation/inhibition | Tim-3+ EM CD8 – inhibition | Functional intermediate (CD16+ CD56br) NK – lytic, cytokine production |
| PD-1+ EM CD4 – activation/inhibition | 41bb+ EM CD8 – costimulation | PD-L1+ functional intermediate NK – inhibition |
| CTLA-4+ EM CD4 – inhibition | Ki67+ EM CD8 – proliferation | PD-1+ functional intermediate NK – activation/inhibition |
| Tim-3+ EM CD4 – inhibition | CD73+ EM CD8 – exhausted/suppressive | Tim-3+ functional intermediate NK – activation/inhibition |
| 41bb+ EM CD4 – costimulation | EMRA (CCR7-CD45RA+) CD8 | Ki67+ functional intermediate NK – proliferation |
| Ki67+ EM CD4 – proliferation | PD-L1+ EMRA CD8 – activation/inhibition | NKp30+ functional intermediate NK – activation |
| CD73+ EM CD4 – exhausted/suppressive | PD-1+ EMRA CD8 – activation/inhibition | NKp46+ functional intermediate NK – activation |
| EMRA (CCR7-CD45RA+) CD4 | CTLA-4+ EMRA CD8 – inhibition | NKG2D+ functional intermediate NK – activation |
| PD-L1+ EMRA CD4 – activation/inhibition | Tim-3+ EMRA CD8 – inhibition | CD226+ functional intermediate NK – adhesion/activation |
| PD-1+ EMRA CD4 – activation/inhibition | 41bb+ EMRA CD8 – costimulation | Immature (CD16- CD56br) NK – cytokine production |
| CTLA-4+ EMRA CD4 – inhibition | Ki67+ EMRA CD8 – proliferation | PD-L1+ immature NK – inhibition |
| Tim-3+ EMRA CD4 – inhibition | CD73+ EMRA CD8 – exhausted/suppressive | PD-1+ immature NK – activation/inhibition |
| 41bb+ EMRA CD4 – costimulation | 3. Total Tregs | Tim-3+ immature NK – activation/inhibition |
| Ki67+ EMRA CD4 – proliferation | PD-L1+ Tregs – activation/inhibition | Ki67+ immature NK – proliferation |
| CD73+ EMRA CD4 – exhausted/suppressive | PD-1+ Tregs – suppression | NKp30+ immature NK – activation |
| CTLA-4+ Tregs – suppression | NKp46+ immature NK – activation | |
| ICOS+ Tregs – suppression | NKG2D+ immature NK – activation | |
| CD45RA+ Tregs – highly expandable in vitro | CD226+ immature NK – adhesion/activation | |
| CD49d- Tregs – suppression | Unconventional (CD16- CD56dim) NK – non-lytic, non-cytokine production | |
| Ki67+ Tregs – proliferation | ||
| CD38+ Tregs – suppression | ||
| HLA-DR+ Tregs – suppression | ||
| 6. Total NK-T | 9. Total MDSC | 10. Total Monocytes |
| PD-L1+ NK-T – inhibition | PD-L1+ MDSC – inhibition | PD-L1+ monocytes – inhibition |
| PD-1+ NK-T – activation/inhibition | PD-1+ MDSC – activation/inhibition | PD-1+ monocytes – activation/inhibition |
| Tim-3+ NK-T – inhibition | CD16+ MDSC – immature/suppression | Classical monocytes – phagocytic |
| Ki67+ NK-T – proliferation | Monocytic (CD14+ CD15-) MDSC | PD-L1+ classical monocytes – inhibition |
| 7. Total cDC | PD-L1+ mMDSC – inhibition | PD-1+ classical monocytes- activation/inhibition |
| PD-L1+ cDC – inhibition | PD-1+ mMDSC – activation/inhibition | Intermediate monocytes phagocytic/proinflammatory |
| PD-1+ cDC – activation/inhibition | CD16+ mMDSC – immature/suppression | PD-L1+ intermediate monocytes – inhibition |
| Tim-3+ cDC – inhibition | Granulocytic (CD14- CD15+) MDSC | PD-1+ intermediate monocytes- activation/inhibition |
| Ki67+ cDC – proliferation | PD-L1+ gMDSC – inhibition | Non-classical monocytes – proinflammatory |
| 8. Total pDC | PD-1+ gMDSC – activation/inhibition | PD-L1+ non-classical monocytes – inhibition |
| PD-L1+ pDC – inhibition | CD16+ gMDSC – immature/suppression | PD-1+ non-classical monocytes- activation/inhibition |
| PD-1+ pDC – activation/inhibition | ||
| Tim-3+ pDC – inhibition | ||
| Ki67+ pDC – proliferation |
Ten parental phenotypes are identified as well as refined subsets of each relating to maturation and function. Expected function based on expression of specific markers within each subset is indicated.
cDC, conventional dendritic cells; CM, central memory; CTLA-4, cytotoxic T lymphocyte-associated protein-4; EM, effector memory; EMRA, terminally differentiated effector memory; gMDSCs, granulocytic myeloid-derived suppressor cells; ICOS, inducible T-cell costimulator; mMDSCs, monocytic MDSCs; NK, natural killer; PD-1, programmed cell death-1; pDC, plasmacytoid DC; PD-L1, programmed cell death ligand-1; Tim-3, T-cell immunoglobulin and mucin domain-3; Tregs, regulatory T cells.