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CISH impairs lysosomal function in activated T cells resulting in mitochondrial DNA release and inflammaging

Nature Aging volume 3, pages 600–616 (2023)Cite this article

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Abstract

Chronic systemic inflammation is one of the hallmarks of the aging immune system. Here we show that activated T cells from older adults contribute to inflammaging by releasing mitochondrial DNA (mtDNA) into their environment due to an increased expression of the cytokine-inducible SH2-containing protein (CISH). CISH targets ATP6V1A, an essential component of the proton pump V-ATPase, for proteasomal degradation, thereby impairing lysosomal function. Impaired lysosomal activity caused intracellular accumulation of multivesicular bodies and amphisomes and the export of their cargos, including mtDNA. CISH silencing in T cells from older adults restored lysosomal activity and prevented amphisomal release. In antigen-specific responses in vivo, CISH-deficient CD4+ T cells released less mtDNA and induced fewer inflammatory cytokines. Attenuating CISH expression may present a promising strategy to reduce inflammation in an immune response of older individuals.

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Fig. 1: Increased expression of CISH in naive CD4+ T-cell responses from older individuals.
Fig. 2: CISH impairs lysosomal activity by promoting proteasomal degradation of ATP6V1A.
Fig. 3: CISH impairs autophagy in activated naive CD4+ T cells from older individuals.
Fig. 4: CISH impairs the clearance of autophagosomes and autolysosomes.
Fig. 5: CISH induces MVB expansion in naive CD4+ T-cell responses from older individuals.
Fig. 6: CISH impairs clearance of damaged mitochondria.
Fig. 7: CISH promotes amphisomal exocytosis of mitochondria into the extracellular environment.
Fig. 8: CISH silencing attenuates mtDNA-induced inflammation in T-cell responses in vivo.

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Data availability

Source data are included with this article. Additional primary experimental data of this study are available from the corresponding author upon request. Sequencing data were obtained from the Sequence Read Archive under accession nos. PRJNA546023 and PRJNA757466.

References

  1. Goronzy, J. J. & Weyand, C. M. Mechanisms underlying T cell ageing. Nat. Rev. Immunol. 19, 573–583 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Nikolich-Zugich, J. The twilight of immunity: emerging concepts in aging of the immune system. Nat. Immunol. 19, 10–19 (2018).

    Article  CAS  PubMed  Google Scholar 

  3. Mittelbrunn, M. & Kroemer, G. Hallmarks of T cell aging. Nat. Immunol. 22, 687–698 (2021).

    Article  CAS  PubMed  Google Scholar 

  4. Franceschi, C., Garagnani, P., Parini, P., Giuliani, C. & Santoro, A. Inflammaging: a new immune-metabolic viewpoint for age-related diseases. Nat. Rev. Endocrinol. 14, 576–590 (2018).

    Article  CAS  PubMed  Google Scholar 

  5. Ferrucci, L. & Fabbri, E. Inflammageing: chronic inflammation in ageing, cardiovascular disease, and frailty. Nat. Rev. Cardiol. 15, 505–522 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Goronzy, J. J. & Weyand, C. M. Successful and maladaptive T cell aging. Immunity 46, 364–378 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Pereira, B. I. et al. Sestrins induce natural killer function in senescent-like CD8+ T cells. Nat. Immunol. 21, 684–694 (2020).

    Article  CAS  PubMed  Google Scholar 

  8. Warrington, K. J., Takemura, S., Goronzy, J. J. & Weyand, C. M. CD4+CD28− T cells in rheumatoid arthritis patients combine features of the innate and adaptive immune systems. Arthritis Rheum. 44, 13–20 (2001).

    Article  CAS  PubMed  Google Scholar 

  9. Henson, S. M., Riddell, N. E. & Akbar, A. N. Properties of end-stage human T cells defined by CD45RA re-expression. Curr. Opin. Immunol. 24, 476–481 (2012).

    Article  CAS  PubMed  Google Scholar 

  10. Kim, C. et al. Activation of miR-21-regulated pathways in immune aging selects against signatures characteristic of memory T cells. Cell Rep. 25, 2148–2162 e2145 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Zhang, H. et al. Aging-associated HELIOS deficiency in naive CD4+ T cells alters chromatin remodeling and promotes effector cell responses. Nat. Immunol. 24, 96–109 (2023).

  12. Kim, C. et al. Histone deficiency and accelerated replication stress in T cell aging. J. Clin. Invest. 131, e143632 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Li, Y. et al. Deficient activity of the nuclease MRE11A induces T cell aging and promotes arthritogenic effector functions in patients with rheumatoid arthritis. Immunity 45, 903–916 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Qi, Q. et al. Defective T memory cell differentiation after Varicella zoster vaccination in older individuals. PLoS Pathog. 12, e1005892 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  15. Lanna, A., Henson, S. M., Escors, D. & Akbar, A. N. The kinase p38 activated by the metabolic regulator AMPK and scaffold TAB1 drives the senescence of human T cells. Nat. Immunol. 15, 965–972 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Mogilenko, D. A. et al. Comprehensive profiling of an aging immune system reveals clonal GZMK+ CD8+ T cells as conserved hallmark of inflammaging. Immunity 54, 99–115 (2021).

  17. Parmigiani, A. et al. Impaired antibody response to influenza vaccine in HIV-infected and uninfected aging women is associated with immune activation and inflammation. PLoS ONE 8, e79816 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  18. Muyanja, E. et al. Immune activation alters cellular and humoral responses to yellow fever 17D vaccine. J. Clin. Invest. 124, 3147–3158 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Berry, M. P. et al. An interferon-inducible neutrophil-driven blood transcriptional signature in human tuberculosis. Nature 466, 973–977 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Fourati, S. et al. Pre-vaccination inflammation and B-cell signalling predict age-related hyporesponse to hepatitis B vaccination. Nat. Commun. 7, 10369 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Vukmanovic-Stejic, M. et al. Enhancement of cutaneous immunity during aging by blocking p38 mitogen-activated protein (MAP) kinase-induced inflammation. J. Allergy Clin. Immunol. 142, 844–856 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Merad, M. & Martin, J. C. Pathological inflammation in patients with COVID-19: a key role for monocytes and macrophages. Nat. Rev. Immunol. 20, 355–362 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Akbar, A. N. & Gilroy, D. W. Aging immunity may exacerbate COVID-19. Science 369, 256–257 (2020).

    Article  CAS  PubMed  Google Scholar 

  24. Doitsh, G. et al. Cell death by pyroptosis drives CD4 T-cell depletion in HIV-1 infection. Nature 505, 509–514 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Li, Y. et al. The DNA repair nuclease MRE11A Functions as a mitochondrial protector and prevents T cell pyroptosis and tissue inflammation. Cell Metab. 30, 477–492 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Jin, J. et al. FOXO1 deficiency impairs proteostasis in aged T cells. Sci. Adv. 6, eaba1808 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Fang, F. et al. Expression of CD39 on activated T cells impairs their survival in older individuals. Cell Rep. 14, 1218–1231 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Matsui, H. et al. Cytosolic dsDNA of mitochondrial origin induces cytotoxicity and neurodegeneration in cellular and zebrafish models of Parkinson’s disease. Nat. Commun. 12, 3101 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Queval, C. J. et al. Mycobacterium tuberculosis controls phagosomal acidification by targeting CISH-mediated signaling. Cell Rep. 20, 3188–3198 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Jadhav, R. R. et al. Epigenetic signature of PD-1+TCF1+ CD8 T cells that act as resource cells during chronic viral infection and respond to PD-1 blockade. Proc. Natl Acad. Sci. USA 116, 14113–14118 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Piessevaux, J., De Ceuninck, L., Catteeuw, D., Peelman, F. & Tavernier, J. Elongin B/C recruitment regulates substrate binding by CIS. J. Biol. Chem. 283, 21334–21346 (2008).

    Article  CAS  PubMed  Google Scholar 

  32. Yoshimura, A., Naka, T. & Kubo, M. SOCS proteins, cytokine signalling and immune regulation. Nat. Rev. Immunol. 7, 454–465 (2007).

    Article  CAS  PubMed  Google Scholar 

  33. Yim, W. W. & Mizushima, N. Lysosome biology in autophagy. Cell Discov. 6, 6 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. He, M. X., McLeod, I. X., Jia, W. & He, Y. W. Macroautophagy in T lymphocyte development and function. Front. Immunol. 3, 22 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Bektas, A. et al. Age-associated changes in human CD4+ T cells point to mitochondrial dysfunction consequent to impaired autophagy. Aging 11, 9234–9263 (2019).

  36. Huotari, J. & Helenius, A. Endosome maturation. EMBO J. 30, 3481–3500 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Yu, Y. R. et al. Disturbed mitochondrial dynamics in CD8+ TILs reinforce T cell exhaustion. Nat. Immunol. 21, 1540–1551 (2020).

  38. Palikaras, K., Lionaki, E. & Tavernarakis, N. Mechanisms of mitophagy in cellular homeostasis, physiology and pathology. Nat. Cell Biol. 20, 1013–1022 (2018).

    Article  CAS  PubMed  Google Scholar 

  39. Ma, K. et al. Mitophagy, mitochondrial homeostasis, and cell fate. Front. Cell Dev. Biol. 8, 467 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  40. Ganesan, D. & Cai, Q. Understanding amphisomes. Biochem. J. 478, 1959–1976 (2021).

    Article  CAS  PubMed  Google Scholar 

  41. Eitan, E., Suire, C., Zhang, S. & Mattson, M. P. Impact of lysosome status on extracellular vesicle content and release. Ageing Res. Rev. 32, 65–74 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Franceschi, C., Garagnani, P., Vitale, G., Capri, M. & Salvioli, S. Inflammaging and ‘Garb-aging’. Trends Endocrinol. Metab. 28, 199–212 (2017).

    Article  CAS  PubMed  Google Scholar 

  43. Pinti, M. et al. Circulating mitochondrial DNA increases with age and is a familiar trait: implications for “inflamm-aging”. Eur. J. Immunol. 44, 1552–1562 (2014).

    Article  CAS  PubMed  Google Scholar 

  44. Riley, J. S. & Tait, S. W. Mitochondrial DNA in inflammation and immunity. EMBO Rep. 21, e49799 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Miller, K. N. et al. Cytoplasmic DNA: sources, sensing, and role in aging and disease. Cell 184, 5506–5526 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Zhang, H., Puleston, D. J. & Simon, A. K. Autophagy and immune senescence. Trends Mol. Med. 22, 671–686 (2016).

    Article  CAS  PubMed  Google Scholar 

  47. Cuervo, A. M. & Macian, F. Autophagy and the immune function in aging. Curr. Opin. Immunol. 29, 97–104 (2014).

    Article  CAS  PubMed  Google Scholar 

  48. Phadwal, K. et al. A novel method for autophagy detection in primary cells: impaired levels of macroautophagy in immunosenescent T cells. Autophagy 8, 677–689 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Raz, Y. et al. Activation-induced autophagy is preserved in CD4+ T-cells in familial longevity. J Gerontol. A Biol. Sci. Med. Sci. 72, 1201–1206 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Alpert, A. et al. A clinically meaningful metric of immune age derived from high-dimensional longitudinal monitoring. Nat. Med. 25, 487–495 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Henson, S. M. et al. p38 signaling inhibits mTORC1-independent autophagy in senescent human CD8+ T cells. J. Clin. Invest. 124, 4004–4016 (2014).

  52. Macian, F. Autophagy in T cell function and aging. Front. Cell Dev. Biol. 7, 213 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  53. Wu, T. et al. The TCF1–Bcl6 axis counteracts type I interferon to repress exhaustion and maintain T cell stemness. Sci. Immunol. 1, eaai8593 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  54. Han, S., Georgiev, P., Ringel, A.E., Sharpe, A.H. & Haigis, M.C. Age-associated remodeling of T cell immunity and metabolism. Cell Metab. 35, 36–55 (2023).

  55. Palmer, D.C. et al. Internal checkpoint regulates T cell neoantigen reactivity and susceptibility to PD1 blockade. Med 3, 682–704.e8 (2022).

  56. Neelapu, S. S. et al. Chimeric antigen receptor T-cell therapy - assessment and management of toxicities. Nat. Rev. Clin. Oncol. 15, 47–62 (2018).

    Article  CAS  PubMed  Google Scholar 

  57. Waldman, A. D., Fritz, J. M. & Lenardo, M. J. A guide to cancer immunotherapy: from T cell basic science to clinical practice. Nat. Rev. Immunol. 20, 651–668 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Mantovani, A., Allavena, P., Sica, A. & Balkwill, F. Cancer-related inflammation. Nature 454, 436–444 (2008).

    Article  CAS  PubMed  Google Scholar 

  59. Arthofer, E. et al. Genetic editing of CISH enhances T cell effector programs independently of immune checkpoint cell surface ligand expression. Preprint at bioRxiv https://doi.org/10.1101/2021.08.17.456714 (2021).

  60. Baixauli, F. et al. Mitochondrial respiration controls lysosomal function during inflammatory T cell responses. Cell Metab. 22, 485–498 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Pence, B. D. et al. Relationship between systemic inflammation and delayed-type hypersensitivity response to Candida antigen in older adults. PLoS ONE 7, e36403 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Watanabe, R. et al. Pyruvate controls the checkpoint inhibitor PD-L1 and suppresses T cell immunity. J. Clin. Invest. 127, 2725–2738 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  63. Teijaro, J. R. et al. Persistent LCMV infection is controlled by blockade of type I interferon signaling. Science 340, 207–211 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Wilson, E. B. et al. Blockade of chronic type I interferon signaling to control persistent LCMV infection. Science 340, 202–207 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. McCausland, M. M. & Crotty, S. Quantitative PCR technique for detecting lymphocytic choriomeningitis virus in vivo. J. Virol. Methods 147, 167–176 (2008).

    Article  CAS  PubMed  Google Scholar 

  66. Steinke, F. C. et al. TCF-1 and LEF-1 act upstream of Th-POK to promote the CD4+ T cell fate and interact with Runx3 to silence Cd4 in CD8+ T cells. Nat. Immunol. 15, 646–656 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank the Mayo Clinic Microscopy and Cell Analysis Core for assistance with flow cytometry (C.A. Moe), cell sorting (Y.H. Han) and TEM (S. Gamb) and R. Ahmed (Emory University) for providing SMARTA mice and LCMV Armstrong. This work was supported by National Institutes of Health grants R01 AR042527, R01 HL117913, R01 AI108906 and R01 HL142068 to C.M.W. and R01 AI108891, R01 AG045779, U19 AI057266 and R01 AI129191 to J.J.G. I.S. was supported by T32AG049672 and by a Glenn Foundation for Medical Research Postdoctoral Fellow. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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  1. These authors contributed equally: Jun Jin, Yunmei Mu.

Authors and Affiliations

  1. Multiscale Research Institute for Complex Systems, Fudan University, Shanghai, China

    Jun Jin

  2. Department of Immunology, Mayo Clinic, Rochester, MN, USA

    Jun Jin, Yunmei Mu, Huimin Zhang, Ines Sturmlechner, Rohit R. Jadhav, Cornelia M. Weyand & Jorg J. Goronzy

  3. Department of Medicine, Stanford University, Stanford, CA, USA

    Jun Jin, Huimin Zhang, Rohit R. Jadhav, Qiong Xia, Cornelia M. Weyand & Jorg J. Goronzy

  4. Department of Medicine, Division of Rheumatology, Mayo Clinic, Rochester, MN, USA

    Chenyao Wang, Cornelia M. Weyand & Jorg J. Goronzy

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Contributions

J.J., Y.M., C.M.W. and J.J.G. designed the study. J.J., Y.M., H.Z., I.S., C.W. and Q.X. performed the experiments. R.J. performed the computational analysis. J.J., Y.M., C.M.W. and J.J.G. analyzed and interpreted the data. J.J., Y.M. and J.J.G. wrote the manuscript with all authors providing feedback.

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Correspondence to Jun Jin or Jorg J. Goronzy.

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Nature Aging thanks Maria Mittelbrunn, Vishwa Dixit, Pedro Rodrigues, and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 TCF1 prevents mtDNA release by promoting lysosomal activity.

a, b, Naïve CD4+ T cells from older 7 females and 7 males were activated for 3 days (a). Cells from 3 young adults were activated in the presence or absence of 10 μM CQ or 5 nM BafA1 (b). mtDNA levels in the culture supernatants were determined. c, Lysosomal acidification was determined by flow cytometry-based analysis of LysoSensor in day-3-stimulated naïve CD4+ T cells comparing 8 young and 8 older individuals. d, Naïve CD4+ T cells from 3 older individuals were transfected with control or CISH siRNA and activated for 3 days. Reduced CISH protein expression after silencing was confirmed by immunoblotting. e, CISH transcripts in naïve CD4+ T cells and naïve CD8+ T cells stimulated for 3 days, in naïve B cells and CD14+ monocytes stimulated for 5 days comparing 6 young and 6 older individuals (left); mtDNA levels in culture supernatants of day-3-activated naïve CD 8+ T cells from 6 older individuals, transfected with siCtrl or siCISH (right). f, TCF protein levels in naïve CD4+ T cells at indicated time points after stimulation comparing 4 young and 4 older individuals. g, Naïve CD4+ T cells from 4 young individuals were transfected with control or TCF7 siRNA and activated for 3 days. Lysosomal acidification (left) and proteolytic activities (right) were determined by flow cytometry-based analysis of LysoSensor and DQ-BSA-treated cells, respectively. h, i, Naïve CD4+ T cells from older individuals were activated for 3 days in the presence of either vehicle or 100 nM of CHIR99021. Lysosomal acidification, proteolytic activities (n = 4) (h) and supernatant mtDNA levels (n = 3) (i) were determined. j, TCF1 and CISH protein in day-3-stimulated naïve CD4+ T cells from 3 older individuals after transfection with pCMV6 control vector or TCF7-expressing vector. Data are presented as mean ±  s.e.m. Comparison by two-tailed unpaired t-test (a, c, e and f) or two-tailed paired t-test (b, d, e and g–j). *P < 0.05, **P < 0.01; NS, not significant.

Source data

Extended Data Fig. 2 Silencing of CISH and ATP6V1A expression in day 3-activated naïve CD4+ T cells.

a, Naïve CD4+ T cells were transfected with control or CISH siRNA and activated for 3 days (n = 3). Lysosomal acidification was determined by flow cytometry-based analysis of LysoSensor-treated cells. b, Naïve CD4+ T cells from a young and an older individual were co-stained with anti-CISH (green) and anti-ATP6V1A Ab (red). Confocal images representative of three independent experiments are shown. c, Proteasome activity in day 3-activated naïve CD4+ T cells from 6 young and 6 older individuals. d, Naïve CD4+ T cells from 3 young individuals were transfected with control or ATP6V1A siRNA and activated for 3 days. Reduced ATP6V1A protein expression after silencing was confirmed by Western blotting. Data are presented as mean ± s.e.m. Comparison by two-tailed paired t-test (a and d) or two-tailed unpaired t-test (c). *P < 0.05; NS, not significant.

Source data

Extended Data Fig. 3 CISH silencing does not affect the number of lysosomes per cell.

Quantitative assessment of lysosome numbers in day 3-activated naïve CD4+ T cells from 5 old individuals after CISH silencing. Image acquisition and analysis were performed by an examiner blinded to the nature of the specimen. Fifteen sections were analyzed for each donor, and mean number per section for each donor is shown NS, not significant. Data are presented as mean ± s.e.m. Comparison by two-tailed unpaired t-test.

Extended Data Fig. 4 Effect of CISH on mitochondria in activated T cells from older adults.

a, COX IV protein expression in day 3-stimulated naïve CD4+ T cells comparing 5 young and 5 older healthy individuals. b, Gating strategy for TMRMhiMtGhi, TMRMloMtGlo and TMRMloMtGhi. TMRMlo cells had reduced cell size (low FSC) and reduced density (low SSC) distinct from apoptotic cells. c, Frequencies of the TMRMloMtGlo population in day 3-activated naïve CD4+ T cells from 4 young and 4 older individuals after indicated gene silencing. Data are presented as mean ± s.e.m. Comparison by two-tailed unpaired t-test (a), one-way ANOVA followed by Tukey’s multiple comparison test (c). *P < 0.05, **P < 0.01 and ***P < 0.001; NS, not significant.

Source data

Extended Data Fig. 5 Confocal image analysis of the effect of CISH on mitochondria and MVBs.

a, b, Naïve CD4+ T cells from a young and an older individual were transfected with indicated siRNA and activated for 3 days. Cells were co-stained with anti-CD63 (as MVB marker) and with anti-LC3B (as autophagosome marker) (a) or anti-COX IV (as mitochondria marker) (b). Confocal images representative of two independent experiments are shown. Scale bars, 1 μm.

Extended Data Fig. 6 CISH-dependent amphisome-rich T cells from older individuals do not show evidence of apoptosis.

a, Naïve CD4+ T cells were transfected with control or indicated siRNA and activated for 3 days (n = 4). Cell apoptosis was examined by staining with Annexin V and 7-AAD. Statistical significance by two-tailed unpaired t-test. NS, not significant. b, Day3-stimulated naïve CD4+ T cells were left untreated or treated with 1 μM Staurosporine for 24 h. Scale bar, 1 μm. Cells were then processed for TEM. Data are presented as mean ± s.e.m.

Extended Data Fig. 7 TCF7 silencing promotes intracellular accumulation of amphisomes.

Naïve CD4+ T cells from young adults were transfected with control or TCF7 siRNA and activated for 3 days. Cells were then harvested and processed for TEM. Representative 30 TEM images (left) and quantitative plots of amphisome numbers in day 3-activated naïve CD4+ T cells from two young individuals after indicated transfection. Data collection and analysis were conducted in a blinded manner. Fifteen cells were analyzed for each donor. Scale bar, 1 μm. Data are presented as mean ± s.e.m. Statistical significance by two-tailed unpaired t-test. ****P < 0.0001.

Extended Data Fig. 8 Effect of CISH on T cell responses in vivo.

a–d, SMARTA cells were transduced and adoptively transferred followed by LCMV infection as described in Fig. 8. Viral RNA levels in the spleen were determined on day 6, n = 10 (a). Frequencies of SMARTA cells with indicated phenotypes in CD4+ donor cells (b–d) on day 6 after LCMV infection (n = 6). e, Mice were reconstituted with shCish or shCtrl retrovirally transduced naïve OT-II CD4+ T cells (n = 3). Frequencies and absolute numbers of TFH and non-TFH OT-II cells in the spleen on day 8 after OVA immunization. Data are presented as mean ± s.e.m. Comparison by two-tailed unpaired t-test (a–e). *P < 0.05; NS, not significant.

Extended Data Fig. 9 Confirmatory studies with a distinct shRNA to silence Cish.

a-d, 1 ×105 Cish shRNA#2 (5′-AAAACAAGTGTTAGAACACAA-3′) or control shRNA retrovirally transduced Amcyan+ naïve SMARTA CD4+ T cells were adoptively transferred into CD45.2+ naïve recipients followed by LCMV infection. a, CISH and ATP6V1A protein expression in transduced cells before adoptive transfer. b, Serum mtDNA copies at day 6 after LCMV infection. c, Serum proinflammatory cytokines levels on day 6 after LCMV infection. d, Gene expression of key inflammatory markers in splenocytes on day 6 after infection. Data are from one experiment with 3–4 mice per group. Data are presented as mean ± s.e.m. Statistical significance by one-way ANOVA followed by Tukey’s multiple comparison test (b and d) or two-tailed unpaired t-test (c). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001; NS, not significant.

Source data

Extended Data Fig. 10

Gating strategy for TFH and non-TFH Amcyan+, successfully transduced cells in the spleen.

Supplementary information

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Supplementary Table 1

Oligonucleotide primer sets used in this study.

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Jin, J., Mu, Y., Zhang, H. et al. CISH impairs lysosomal function in activated T cells resulting in mitochondrial DNA release and inflammaging. Nat Aging 3, 600–616 (2023). https://doi.org/10.1038/s43587-023-00399-w

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