This study discusses substantive advances in T cell proliferation analysis, with the aim to provoke a re-evaluation of the generally-held view that Ki-67 is a reliable proliferation marker per se , and to offer a more sensitive and effective method for T cell cycle analysis, with informative examples in mouse and human settings. We summarize recent experimental work from our labs showing that, by Ki-67/DNA dual staining and refined flow cytometric methods, we were able to identify T cells in the S-G 2 /M phases of the cell-cycle in the peripheral blood (collectively termed “T Double S” for T cells in S -phase in S anguine : in short “T DS ” cells). Without our refinement, such cells may be excluded from conventional lymphocyte analyses. Specifically, we analyzed clonal expansion of antigen-specific CD8 T cells in vaccinated mice, and demonstrated the potential of T DS cells to reflect immune dynamics in human blood samples from healthy donors, and patients with type 1 diabetes, infectious mononucleosis, and COVID-19. The Ki-67/DNA dual staining, or T DS assay, provides a reliable approach by which human peripheral blood can be used to reflect the dynamics of human lymphocytes, rather than providing mere steady-state phenotypic snapshots. The method does not require highly sophisticated “-omics” capabilities, so it should be widely-applicable to health care in diverse settings. Furthermore, our results argue that the T DS assay can provide a window on immune dynamics in extra-lymphoid tissues, a long-sought potential of peripheral blood monitoring, for example in relation to organ-specific autoimmune diseases and infections, and cancer immunotherapy.
【저자키워드】 T cells, flow cytometry, cell cycle, Ki-67, DNA dye, 【초록키워드】 COVID-19, Cancer immunotherapy, Type 1 diabetes, Health care, Lymphocytes, T cells, diabetes, immune, cell cycle, Ki-67, Antigen, Peripheral blood, Autoimmune disease, lymphocyte, T cell, Health, infections, cells, mice, Patient, Autoimmune diseases, Care, CD8 T cells, marker, Analysis, CD8 T cell, proliferation, dual, clonal expansion, infectious mononucleosis, tissues, healthy donors, blood sample, window, phenotypic, antigen-specific CD8 T cells, human peripheral blood, re-evaluation, staining, experimental work, offer, lab, approach, effective, Cell, T cell proliferation, flow cytometric, analyzed, identify, example, can be used, provide, demonstrated, excluded, analyses, organ-specific, provoke, 【제목키워드】 Potential,