Differentiated somatic mammalian cells putatively exhibit species-specific division limits that impede cancer but may constrain lifespans1,2,3. To provide immunity, transiently stimulated CD8+ T cells undergo unusually rapid bursts of numerous cell divisions, and then form quiescent long-lived memory cells that remain poised to reproliferate following subsequent immunological challenges. Here we addressed whether T cells are intrinsically constrained by chronological or cell-division limits. We activated mouse T cells in vivo using acute heterologous prime–boost–boost vaccinations4, transferred expanded cells to new mice, and then repeated this process iteratively. Over 10 years (greatly exceeding the mouse lifespan)5 and 51 successive immunizations, T cells remained competent to respond to vaccination. Cells required sufficient rest between stimulation events. Despite demonstrating the potential to expand the starting population at least 1040-fold, cells did not show loss of proliferation control and results were not due to contamination with young cells. Persistent stimulation by chronic infections or cancer can cause T cell proliferative senescence, functional exhaustion and death6. We found that although iterative acute stimulations also induced sustained expression and epigenetic remodelling of common exhaustion markers (including PD1, which is also known as PDCD1, and TOX) in the cells, they could still proliferate, execute antimicrobial functions and form quiescent memory cells. These observations provide a model to better understand memory cell differentiation, exhaustion, cancer and ageing, and show that functionally competent T cells can retain the potential for extraordinary population expansion and longevity well beyond their organismal lifespan.
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We thank members of the laboratories of D.M. and V.V. for helpful discussions. We thank the University of Minnesota Flow Cytometry resource for cell sorting (J. Motl, T. Martin and R. Arora). This study was supported by National Institutes of Health grants R01 AI084913, R01 AI146032 (D.M.) and T32HL007741 (A.G.S.), and Swiss National Science Foundation grant P2BSP3_200187 (M.K.).
The authors declare no competing interests.
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Extended data figures and tables
a) CD45.1+ 48° ISTC memory CD8 T cells were transferred to naïve mice, followed by three heterologous prime-boost-boost immunizations. Non-lymphoid tissues were analyzed 48 days after the 51° boost. Flow cytometry plots are gated on live CD8a+ lymphocytes that were not stained by intravascular in vivo antibody labeling. b) CD69 expression on endogenous 3° cells (CD45.1-) and 48° memory CD8 T cells (CD45.1+).Plots concatenated from four mice and experiment is representative of three similar experiments with similar results.
Gene set enrichment analysis was performed on differentially expressed genes between 45° ISTCs and primary memory cells using the category “Biological process” of the Gene Ontology database. The top 20 upregulated (activated) and top 20 downregulated (suppressed) pathways in ISTCs are shown. Statistical significance was determined using Over-Representation analysis.
Extended Data Fig. 3 A single naïve CD8 T cell shows the potential to produce >1040 memory cell progeny.
a) Schematic showing estimate of cell expansion potential after 51 HPBB immunizations. HPBB: three successive Heterologous Prime, Boost, Boost immunizations. b) Schematic comparing the volume of earth with the theoretical volume of memory T cells implied by the calculated proliferation potential of a naïve CD8 T cell.
This file lists the differentially expressed genes between 45° cells and 3° cells, 1° or naive CD8+ T cells. Empirical Bayes moderation was used to test for significance.
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Soerens, A.G., Künzli, M., Quarnstrom, C.F. et al. Functional T cells are capable of supernumerary cell division and longevity. Nature 614, 762–766 (2023). https://doi.org/10.1038/s41586-022-05626-9
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