Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq

IntroductionThe functional programs of CD4+ T helper (Th) cell clones play a central role in shaping immune responses to different challenges. While advances in single-cell RNA sequencing (scRNA-Seq) have significantly improved our understanding of the diversity of Th cells, the relationship between...

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Main Authors: Daniil K. Lukyanov, Valeriia V. Kriukova, Kristin Ladell, Irina A. Shagina, Dmitry B. Staroverov, Bella E. Minasian, Anna S. Fedosova, Pavel Shelyakin, Oleg N. Suchalko, Alexander Y. Komkov, Konstantin A. Blagodatskikh, Kelly L. Miners, Olga V. Britanova, Andre Franke, David A. Price, Dmitry M. Chudakov
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Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1536302/full
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author Daniil K. Lukyanov
Daniil K. Lukyanov
Valeriia V. Kriukova
Kristin Ladell
Irina A. Shagina
Irina A. Shagina
Dmitry B. Staroverov
Dmitry B. Staroverov
Bella E. Minasian
Anna S. Fedosova
Pavel Shelyakin
Oleg N. Suchalko
Alexander Y. Komkov
Konstantin A. Blagodatskikh
Kelly L. Miners
Olga V. Britanova
Olga V. Britanova
Olga V. Britanova
Andre Franke
David A. Price
David A. Price
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
author_facet Daniil K. Lukyanov
Daniil K. Lukyanov
Valeriia V. Kriukova
Kristin Ladell
Irina A. Shagina
Irina A. Shagina
Dmitry B. Staroverov
Dmitry B. Staroverov
Bella E. Minasian
Anna S. Fedosova
Pavel Shelyakin
Oleg N. Suchalko
Alexander Y. Komkov
Konstantin A. Blagodatskikh
Kelly L. Miners
Olga V. Britanova
Olga V. Britanova
Olga V. Britanova
Andre Franke
David A. Price
David A. Price
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
author_sort Daniil K. Lukyanov
collection DOAJ
description IntroductionThe functional programs of CD4+ T helper (Th) cell clones play a central role in shaping immune responses to different challenges. While advances in single-cell RNA sequencing (scRNA-Seq) have significantly improved our understanding of the diversity of Th cells, the relationship between scRNA-Seq clusters and the traditionally characterized Th subsets remains ambiguous.MethodsIn this study, we introduce TCR-Track, a method leveraging immune repertoire data to map phenotypically sorted Th subsets onto scRNA-Seq profiles.Results and discussionThis approach accurately positions the Th1, Th1-17, Th17, Th22, Th2a, Th2, T follicular helper (Tfh), and regulatory T-cell (Treg) subsets, outperforming mapping based on CITE-Seq. Remarkably, the mapping is tightly focused on specific scRNA-Seq clusters, despite 4-year interval between subset sorting and the effector CD4+ scRNA-Seq experiment. These findings highlight the intrinsic program stability of Th clones circulating in peripheral blood. Repertoire overlap analysis at the scRNA-Seq level confirms that the circulating Th1, Th2, Th2a, Th17, Th22, and Treg subsets are clonally independent. However, a significant clonal overlap between the Th1 and cytotoxic CD4+ T-cell clusters suggests that cytotoxic CD4+ T cells differentiate from Th1 clones. In addition, this study resolves a longstanding ambiguity: we demonstrate that, while CCR10+ Th cells align with a specific Th22 scRNA-Seq cluster, CCR10−CCR6+CXCR3−CCR4+ cells, typically classified as Th17, represent a mixture of bona fide Th17 cells and clonally unrelated CCR10low Th22 cells. The clear distinction between the Th17 and Th22 subsets should influence the development of vaccine- and T-cell-based therapies. Furthermore, we show that severe acute SARS-CoV-2 infection induces systemic type 1 interferon (IFN) activation of naive Th cells. An increased proportion of effector IFN-induced Th cells is associated with a moderate course of the disease but remains low in critical COVID-19 cases. Using integrated scRNA-Seq, TCR-Track, and CITE-Seq data from 122 donors, we provide a comprehensive Th scRNA-Seq reference that should facilitate further investigation of Th subsets in fundamental and clinical studies.
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spelling doaj-art-a2570c81a9a9422faeb85902f7785ab92025-08-20T03:05:43ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-04-011610.3389/fimmu.2025.15363021536302Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-SeqDaniil K. Lukyanov0Daniil K. Lukyanov1Valeriia V. Kriukova2Kristin Ladell3Irina A. Shagina4Irina A. Shagina5Dmitry B. Staroverov6Dmitry B. Staroverov7Bella E. Minasian8Anna S. Fedosova9Pavel Shelyakin10Oleg N. Suchalko11Alexander Y. Komkov12Konstantin A. Blagodatskikh13Kelly L. Miners14Olga V. Britanova15Olga V. Britanova16Olga V. Britanova17Andre Franke18David A. Price19David A. Price20Dmitry M. Chudakov21Dmitry M. Chudakov22Dmitry M. Chudakov23Dmitry M. Chudakov24Dmitry M. Chudakov25Center for Molecular and Cellular Biology, Moscow, RussiaGenomics of Adaptive Immunity Department, Institute of Bioorganic Chemistry, Moscow, RussiaInstitute of Clinical Molecular Biology, Kiel University, Kiel, GermanyDivision of Infection and Immunity, Cardiff University School of Medicine, University Hospital of Wales, Cardiff, United KingdomGenomics of Adaptive Immunity Department, Institute of Bioorganic Chemistry, Moscow, RussiaInstitute of Translational Medicine, Pirogov Russian National Research Medical University, Moscow, RussiaGenomics of Adaptive Immunity Department, Institute of Bioorganic Chemistry, Moscow, RussiaInstitute of Translational Medicine, Pirogov Russian National Research Medical University, Moscow, RussiaAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesInstitute of Translational Medicine, Pirogov Russian National Research Medical University, Moscow, RussiaDivision of Infection and Immunity, Cardiff University School of Medicine, University Hospital of Wales, Cardiff, United KingdomGenomics of Adaptive Immunity Department, Institute of Bioorganic Chemistry, Moscow, RussiaInstitute of Translational Medicine, Pirogov Russian National Research Medical University, Moscow, RussiaAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesInstitute of Clinical Molecular Biology, Kiel University, Kiel, GermanyDivision of Infection and Immunity, Cardiff University School of Medicine, University Hospital of Wales, Cardiff, United KingdomSystems Immunity Research Institute, Cardiff University School of Medicine, University Hospital of Wales, Cardiff, United KingdomCenter for Molecular and Cellular Biology, Moscow, RussiaGenomics of Adaptive Immunity Department, Institute of Bioorganic Chemistry, Moscow, RussiaInstitute of Translational Medicine, Pirogov Russian National Research Medical University, Moscow, RussiaAbu Dhabi Stem Cell Center, Al Muntazah, United Arab EmiratesDepartment of Molecular Medicine, Central European Institute of Technology, Brno, CzechiaIntroductionThe functional programs of CD4+ T helper (Th) cell clones play a central role in shaping immune responses to different challenges. While advances in single-cell RNA sequencing (scRNA-Seq) have significantly improved our understanding of the diversity of Th cells, the relationship between scRNA-Seq clusters and the traditionally characterized Th subsets remains ambiguous.MethodsIn this study, we introduce TCR-Track, a method leveraging immune repertoire data to map phenotypically sorted Th subsets onto scRNA-Seq profiles.Results and discussionThis approach accurately positions the Th1, Th1-17, Th17, Th22, Th2a, Th2, T follicular helper (Tfh), and regulatory T-cell (Treg) subsets, outperforming mapping based on CITE-Seq. Remarkably, the mapping is tightly focused on specific scRNA-Seq clusters, despite 4-year interval between subset sorting and the effector CD4+ scRNA-Seq experiment. These findings highlight the intrinsic program stability of Th clones circulating in peripheral blood. Repertoire overlap analysis at the scRNA-Seq level confirms that the circulating Th1, Th2, Th2a, Th17, Th22, and Treg subsets are clonally independent. However, a significant clonal overlap between the Th1 and cytotoxic CD4+ T-cell clusters suggests that cytotoxic CD4+ T cells differentiate from Th1 clones. In addition, this study resolves a longstanding ambiguity: we demonstrate that, while CCR10+ Th cells align with a specific Th22 scRNA-Seq cluster, CCR10−CCR6+CXCR3−CCR4+ cells, typically classified as Th17, represent a mixture of bona fide Th17 cells and clonally unrelated CCR10low Th22 cells. The clear distinction between the Th17 and Th22 subsets should influence the development of vaccine- and T-cell-based therapies. Furthermore, we show that severe acute SARS-CoV-2 infection induces systemic type 1 interferon (IFN) activation of naive Th cells. An increased proportion of effector IFN-induced Th cells is associated with a moderate course of the disease but remains low in critical COVID-19 cases. Using integrated scRNA-Seq, TCR-Track, and CITE-Seq data from 122 donors, we provide a comprehensive Th scRNA-Seq reference that should facilitate further investigation of Th subsets in fundamental and clinical studies.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1536302/fullhelper T cell subsetsscRNA-SeqscTCR-seqimmune repertoiresT cell memoryTh22
spellingShingle Daniil K. Lukyanov
Daniil K. Lukyanov
Valeriia V. Kriukova
Kristin Ladell
Irina A. Shagina
Irina A. Shagina
Dmitry B. Staroverov
Dmitry B. Staroverov
Bella E. Minasian
Anna S. Fedosova
Pavel Shelyakin
Oleg N. Suchalko
Alexander Y. Komkov
Konstantin A. Blagodatskikh
Kelly L. Miners
Olga V. Britanova
Olga V. Britanova
Olga V. Britanova
Andre Franke
David A. Price
David A. Price
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Dmitry M. Chudakov
Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
Frontiers in Immunology
helper T cell subsets
scRNA-Seq
scTCR-seq
immune repertoires
T cell memory
Th22
title Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
title_full Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
title_fullStr Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
title_full_unstemmed Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
title_short Repertoire-based mapping and time-tracking of T helper cell subsets in scRNA-Seq
title_sort repertoire based mapping and time tracking of t helper cell subsets in scrna seq
topic helper T cell subsets
scRNA-Seq
scTCR-seq
immune repertoires
T cell memory
Th22
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1536302/full
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