Early transcriptional and epigenetic divergence of CD8+ T cells responding to acute versus chronic infection

During a microbial infection, responding CD8+ T cells give rise to effector cells that provide acute host defense and memory cells that provide sustained protection. An alternative outcome is exhaustion, a state of T cell dysfunction that occurs in the context of chronic infections and cancer. Although it is evident that exhausted CD8+ T (TEX) cells are phenotypically and molecularly distinct from effector and memory CD8+ T cells, the factors regulating the earliest events in the differentiation process of TEX cells remain incompletely understood. Here, we performed single-cell RNA-sequencing and single-cell ATAC-sequencing of CD8+ T cells responding to LCMV-Armstrong (LCMV-Arm) or LCMV-Clone 13 (LCMV-Cl13), which result in acute or chronic infections, respectively. Compared to CD8+ T cells that had undergone their first division in response to LCMV-Arm (Div1ARM) cells, CD8+ T cells that had undergone their first division in response to LCMV-Cl13 (Div1CL13) expressed higher levels of genes encoding transcription factors previously associated with exhaustion, along with higher levels of Ezh2, the catalytic component of the Polycomb Repressive Complex 2 (PRC2) complex, which mediates epigenetic silencing. Modulation of Ezh2 resulted in altered expression of exhaustion-associated molecules by CD8+ T cells responding to LCMV-Cl13, though the specific cellular and infectious contexts, rather than simply the level of Ezh2 expression, likely determine the eventual outcome. Taken together, these findings suggest that the differentiation paths of CD8+ T cells responding to acute versus chronic infections may diverge earlier than previously appreciated.

[1]  V. Buchholz,et al.  MYB orchestrates T cell exhaustion and response to checkpoint inhibition , 2022, Nature.

[2]  John T. Chang,et al.  Tissue-resident memory CD8+ T cells possess unique transcriptional, epigenetic and functional adaptations to different tissue environments , 2022, Nature Immunology.

[3]  John T. Chang,et al.  The long noncoding RNA Malat1 regulates CD8+ T cell differentiation by mediating epigenetic repression , 2022, The Journal of experimental medicine.

[4]  C. Lareau,et al.  Single-cell chromatin state analysis with Signac , 2021, Nature Methods.

[5]  E. Wherry,et al.  Epigenetic scarring of exhausted T cells hinders memory differentiation upon eliminating chronic antigenic stimulation , 2021, Nature Immunology.

[6]  E. Wherry,et al.  Memory T-Cell Heterogeneity and Terminology. , 2021, Cold Spring Harbor perspectives in biology.

[7]  Anushya Muruganujan,et al.  PANTHER version 16: a revised family classification, tree-based classification tool, enhancer regions and extensive API , 2020, Nucleic Acids Res..

[8]  David Chisanga,et al.  Early precursor T cells establish and propagate T cell exhaustion in chronic infection , 2020, Nature Immunology.

[9]  John T. Chang,et al.  Heterogenous Populations of Tissue-Resident CD8+ T Cells Are Generated in Response to Infection and Malignancy. , 2020, Immunity.

[10]  John T. Chang,et al.  Early precursors and molecular determinants of tissue-resident memory CD8+ T lymphocytes revealed by single-cell RNA sequencing , 2020, Science Immunology.

[11]  E. Wherry,et al.  Developmental Relationships of Four Exhausted CD8+ T Cell Subsets Reveals Underlying Transcriptional and Epigenetic Landscape Control Mechanisms. , 2020, Immunity.

[12]  Lun Song,et al.  Novel antibody epitopes dominate the antigenicity of spike glycoprotein in SARS-CoV-2 compared to SARS-CoV , 2020, Cellular & Molecular Immunology.

[13]  G. Freeman,et al.  Proliferating Transitory T Cells with an Effector-like Transcriptional Signature Emerge from PD-1+ Stem-like CD8+ T Cells during Chronic Infection. , 2019, Immunity.

[14]  S. Berger,et al.  TCF-1-Centered Transcriptional Network Drives an Effector versus Exhausted CD8 T Cell-Fate Decision. , 2019, Immunity.

[15]  Fabian J Theis,et al.  Generalizing RNA velocity to transient cell states through dynamical modeling , 2019, Nature Biotechnology.

[16]  E. Wherry,et al.  Defining ‘T cell exhaustion’ , 2019, Nature Reviews Immunology.

[17]  Elisabeth F. Heuston,et al.  Single-Cell RNA-Seq Reveals TOX as a Key Regulator of CD8+ T cell Persistence in Chronic Infection , 2019, Nature Immunology.

[18]  M. Delorenzi,et al.  TOX reinforces the phenotype and longevity of exhausted T cells in chronic viral infection , 2019, Nature.

[19]  Yong Liu,et al.  TOX is a critical regulator of tumour-specific T cell differentiation , 2019, Nature.

[20]  S. Berger,et al.  TOX transcriptionally and epigenetically programs CD8+ T cell exhaustion , 2019, Nature.

[21]  P. Hogan,et al.  TOX and TOX2 transcription factors cooperate with NR4A transcription factors to impose CD8+ T cell exhaustion , 2019, Proceedings of the National Academy of Sciences.

[22]  W. Telford,et al.  miR-155 harnesses Phf19 to potentiate cancer immunotherapy through epigenetic reprogramming of CD8+ T cell fate , 2019, Nature Communications.

[23]  E. Wherry,et al.  CD8 T Cell Exhaustion During Chronic Viral Infection and Cancer. , 2019, Annual review of immunology.

[24]  Howard Y. Chang,et al.  Massively parallel single-cell chromatin landscapes of human immune cell development and intratumoral T cell exhaustion , 2019, Nature Biotechnology.

[25]  A. Oxenius,et al.  Modulation of asymmetric cell division as a mechanism to boost CD8+ T cell memory , 2019, Science Immunology.

[26]  Olga Tanaseichuk,et al.  Metascape provides a biologist-oriented resource for the analysis of systems-level datasets , 2019, Nature Communications.

[27]  Lu Huang,et al.  Integrative network modeling reveals mechanisms underlying T cell exhaustion , 2019, Scientific Reports.

[28]  Jianjun Hu,et al.  The Transcription Factor TCF1 Preserves the Effector Function of Exhausted CD8 T Cells During Chronic Viral Infection , 2019, Front. Immunol..

[29]  A. Yoshimura,et al.  Nr4a transcription factors limit CAR T cell function in solid tumors , 2019, Nature.

[30]  Erik Sundström,et al.  RNA velocity of single cells , 2018, Nature.

[31]  Fabian J Theis,et al.  SCANPY: large-scale single-cell gene expression data analysis , 2018, Genome Biology.

[32]  W. Shi,et al.  Transcription Factor IRF4 Promotes CD8+ T Cell Exhaustion and Limits the Development of Memory‐like T Cells during Chronic Infection , 2017, Immunity.

[33]  Christina S. Leslie,et al.  Chromatin states define tumor-specific T cell dysfunction and reprogramming , 2017, Nature.

[34]  Steven H. Kleinstein,et al.  Polycomb Repressive Complex 2‐Mediated Chromatin Repression Guides Effector CD8+ T Cell Terminal Differentiation and Loss of Multipotency , 2017, Immunity.

[35]  A. Hoffmann,et al.  Exhaustion-associated regulatory regions in CD8+ tumor-infiltrating T cells , 2017, Proceedings of the National Academy of Sciences.

[36]  John T. Chang,et al.  Early transcriptional and epigenetic regulation of CD8+ T cell differentiation revealed by single-cell RNA-seq , 2017, Nature Immunology.

[37]  Todd M. Allen,et al.  The epigenetic landscape of T cell exhaustion , 2016, Science.

[38]  S. Berger,et al.  Epigenetic stability of exhausted T cells limits durability of reinvigoration by PD-1 blockade , 2016, Science.

[39]  A. Bhandoola,et al.  CD8+ T Lymphocyte Self-Renewal during Effector Cell Determination. , 2016, Cell reports.

[40]  Matheus C. Bürger,et al.  Defining CD8+ T cells that provide the proliferative burst after PD-1 therapy , 2016, Nature.

[41]  Sandra P. Calderon-Copete,et al.  T Cell Factor 1-Expressing Memory-like CD8(+) T Cells Sustain the Immune Response to Chronic Viral Infections. , 2016, Immunity.

[42]  Greg M. Delgoffe,et al.  Asymmetric inheritance of mTORC1 kinase activity during division dictates CD8 T cell differentiation , 2016, Nature Immunology.

[43]  D. Green,et al.  Metabolic Maintenance of Cell Asymmetry following Division in Activated T Lymphocytes , 2016, Nature.

[44]  David C. Norris,et al.  Integrated genome browser: visual analytics platform for genomics , 2015, bioRxiv.

[45]  E. Wherry,et al.  Molecular and cellular insights into T cell exhaustion , 2015, Nature Reviews Immunology.

[46]  H. Lähdesmäki,et al.  The transcription factor NFAT promotes exhaustion of activated CD8⁺ T cells. , 2015, Immunity.

[47]  John T. Chang,et al.  Molecular regulation of effector and memory T cell differentiation , 2014, Nature Immunology.

[48]  John T. Chang,et al.  Early specification of CD8+ T lymphocyte fates during adaptive immunity revealed by single-cell gene expression analyses , 2014, Nature Immunology.

[49]  R. Schreiber,et al.  Persistent LCMV Infection Is Controlled by Blockade of Type I Interferon Signaling , 2013, Science.

[50]  Burton E. Barnett,et al.  Progenitor and Terminal Subsets of CD8+ T Cells Cooperate to Contain Chronic Viral Infection , 2012, Science.

[51]  Barbara Hausmann,et al.  T cell affinity regulates asymmetric division, effector cell differentiation, and tissue pathology. , 2012, Immunity.

[52]  ENCODEConsortium,et al.  An Integrated Encyclopedia of DNA Elements in the Human Genome , 2012, Nature.

[53]  R. Flavell,et al.  Plasmacytoid dendritic cells are productively infected and activated through TLR-7 early after arenavirus infection. , 2012, Cell host & microbe.

[54]  E. Wherry,et al.  Progressive Loss of Memory T Cell Potential and Commitment to Exhaustion during Chronic Viral Infection , 2012, Journal of Virology.

[55]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[56]  E John Wherry,et al.  T cell exhaustion , 2011 .

[57]  John T. Chang,et al.  Asymmetric proteasome segregation as a mechanism for unequal partitioning of the transcription factor T-bet during T lymphocyte division. , 2011, Immunity.

[58]  Clifford A. Meyer,et al.  Model-based Analysis of ChIP-Seq (MACS) , 2008, Genome Biology.

[59]  John T. Chang,et al.  Asymmetric T Lymphocyte Division in the Initiation of Adaptive Immune Responses , 2007, Science.

[60]  K. Campbell,et al.  Differences in Affinity of Binding of Lymphocytic Choriomeningitis Virus Strains to the Cellular Receptor α-Dystroglycan Correlate with Viral Tropism and Disease Kinetics , 2001, Journal of Virology.

[61]  Andreas Holz,et al.  Immunosuppression and Resultant Viral Persistence by Specific Viral Targeting of Dendritic Cells , 2000, The Journal of experimental medicine.

[62]  P. Borrow,et al.  Virus-induced immunosuppression: immune system-mediated destruction of virus-infected dendritic cells results in generalized immune suppression , 1995, Journal of virology.