GZMK⁺CD8⁺ T cells target a specific acinar cell type in Sjögren's disease

Thomas J F Pranzatelli¹, Paola Perez², Anson Ku³, Bruno Matuck⁴, Khoa Huynh⁵, Shunsuke Sakai⁶, Mehdi Abed², Shyh-Ing Jang², Eiko Yamada², Kalie Dominick², Zara Ahmed², Amanda J Oliver⁷, Rachael Bogle⁸, Quinn T Easter⁴, Alan N Baer⁹, Eileen Pelayo⁹, Zohreh Khavandgar¹⁰, David E Kleiner¹¹, M Teresa Magone¹², Sarthak Gupta¹³, Christopher Lessard¹⁴, A Darise Farris¹⁵, Peter D Burbelo¹⁶, Daniel Martin¹⁷, Robert J Morell¹⁷, Changyu Zheng¹⁷, Nicholas Rachmaninoff¹⁸, Jose Maldonado-Ortiz¹⁹, Katarzyna M Tyc⁵, Xufeng Qu²⁰, Marit Aure²¹, Mohammad H Dezfulian²², Ross Lake²³, Sarah A Teichmann²⁴, Daniel L Barber⁶, Lam C Tsoi⁸, Adam G Sowalsky³, Jinze Liu⁵, Johann Gudjonsson⁸, Kevin M Byrd²⁵, Philip L F Johnson¹⁸, John A Chiorini¹⁶, Blake M Warner²⁶

Affiliations

¹ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Department of Biology, University of Maryland, College Park, Maryland, USA.
² Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
³ Laboratory of Genitourinary Cancer Pathogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
⁴ Lab of Oral & Craniofacial Innovation, Department of Innovation & Technology Research, ADA Science & Research Institute, Gaithersburg, Maryland, USA.
⁵ Department of Biostatistics, Virginia Commonwealth University, Richmond, Virginia, USA.
⁶ T-lymphocyte Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
⁷ Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.
⁸ Department of Dermatology, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, USA.
⁹ Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹⁰ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹¹ Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
¹² Consult Services Section, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
¹³ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Lupus Clinical Trials Unit, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA.
¹⁴ Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
¹⁵ Arthritis & Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
¹⁶ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹⁷ Genomics and Computational Biology Core, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA.
¹⁸ Department of Biology, University of Maryland, College Park, Maryland, USA.
¹⁹ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
²⁰ Massey Cancer Center, Virginia Commonwealth University, Richmond, Virginia, USA.
²¹ Matrix and Morphogenesis Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
²² Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
²³ Laboratory of Genitourinary Cancer Pathogenesis (LCGP) Microscopy Core Facility, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
²⁴ Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK; Ensocell Therapeutics, BioData Innovation Centre, Wellcome Genome Campus, Cambridge, UK; Theory of Condensed Matter, Cavendish Laboratory/Department of Physics, University of Cambridge, Cambridge, UK; Department of Medicine, University of Cambridge, Cambridge, UK; CIFAR Macmillan Multi-scale Human Program, CIFAR, Toronto, Ontario, Canada.
²⁵ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Lab of Oral & Craniofacial Innovation, Department of Innovation & Technology Research, ADA Science & Research Institute, Gaithersburg, Maryland, USA; Department of Oral and Craniofacial Molecular Biology, Philips Institute for Oral Health Research, Virginia Commonwealth University, Richmond, Virginia, USA; VCU Massey Comprehensive Cancer Center, Bioinformatics Shared Resource Core, Virginia Commonwealth University, Richmond, Virginia, USA.
²⁶ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA. Electronic address: blake.warner@nih.gov.

PMID: 41162286
PMCID: PMC12854051 (available on 2026-10-28)
DOI: 10.1016/j.ard.2025.08.014

GZMK⁺CD8⁺ T cells target a specific acinar cell type in Sjögren's disease

Thomas J F Pranzatelli et al. Ann Rheum Dis. 2025.

. 2025 Oct 28:S0003-4967(25)04313-4.

doi: 10.1016/j.ard.2025.08.014. Online ahead of print.

Authors

Affiliations

¹ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Department of Biology, University of Maryland, College Park, Maryland, USA.
² Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
³ Laboratory of Genitourinary Cancer Pathogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
⁴ Lab of Oral & Craniofacial Innovation, Department of Innovation & Technology Research, ADA Science & Research Institute, Gaithersburg, Maryland, USA.
⁵ Department of Biostatistics, Virginia Commonwealth University, Richmond, Virginia, USA.
⁶ T-lymphocyte Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
⁷ Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.
⁸ Department of Dermatology, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, USA.
⁹ Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹⁰ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹¹ Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
¹² Consult Services Section, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
¹³ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Lupus Clinical Trials Unit, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA.
¹⁴ Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
¹⁵ Arthritis & Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
¹⁶ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
¹⁷ Genomics and Computational Biology Core, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA.
¹⁸ Department of Biology, University of Maryland, College Park, Maryland, USA.
¹⁹ Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
²⁰ Massey Cancer Center, Virginia Commonwealth University, Richmond, Virginia, USA.
²¹ Matrix and Morphogenesis Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
²² Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
²³ Laboratory of Genitourinary Cancer Pathogenesis (LCGP) Microscopy Core Facility, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
²⁴ Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK; Ensocell Therapeutics, BioData Innovation Centre, Wellcome Genome Campus, Cambridge, UK; Theory of Condensed Matter, Cavendish Laboratory/Department of Physics, University of Cambridge, Cambridge, UK; Department of Medicine, University of Cambridge, Cambridge, UK; CIFAR Macmillan Multi-scale Human Program, CIFAR, Toronto, Ontario, Canada.
²⁵ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Lab of Oral & Craniofacial Innovation, Department of Innovation & Technology Research, ADA Science & Research Institute, Gaithersburg, Maryland, USA; Department of Oral and Craniofacial Molecular Biology, Philips Institute for Oral Health Research, Virginia Commonwealth University, Richmond, Virginia, USA; VCU Massey Comprehensive Cancer Center, Bioinformatics Shared Resource Core, Virginia Commonwealth University, Richmond, Virginia, USA.
²⁶ Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA; Sjögren's Clinical Investigations Team, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA. Electronic address: blake.warner@nih.gov.

PMID: 41162286
PMCID: PMC12854051 (available on 2026-10-28)
DOI: 10.1016/j.ard.2025.08.014

Abstract

Objectives: Sjögren's disease (SjD) is a systemic autoimmune disorder characterized by dysfunction of exocrine glands, particularly the salivary and lacrimal glands, with no clear etiology or effective therapy. This study explores the complex interplay of varied cell types in the salivary glands and their role in the pathology of Sjögren's disease.

Methods: Utilizing single-cell and spatial transcriptomics alongside spatial immunophenotyping to analyze human minor salivary glands, we developed a comprehensive understanding of the cellular landscape of non-SjD salivary glands and how that landscape changes in SjD patients. In vitro cellular assays and novel patient-derived primary epithelial cells were co-cultured with autologous T cells to confirm effector states and the delivery and effect of disease-associated granzymes.

Results: We identified previously unrecognized heterogeneity among acinar cells, including a PRR4⁺CST3⁺WFDC2⁻ seromucous acinar population that is selectively lost in Sjögren's disease. Expression and organizational changes were linked to clinical features: (i) T cells in the glands of SSA⁺, high-focus score patients showed increased transcriptional signatures of activation, antigen presentation, and apoptosis resistance compared with patients with mild or moderate disease, and (ii) patients with low immune infiltration exhibited distinct epithelial organization. Notably, GZMK⁺CD8⁺ T cells, which accumulate with disease severity, displayed a cytotoxic transcriptional program, degranulated upon stimulation ex vivo, and localized spatially with immune-engaged epithelial cells. Functional assays demonstrated that GZMK activates interferon signaling in vitro, and autologous co-cultures of patient-derived T cells and epithelial cells validated these findings.

Conclusions: Using single-cell and spatial transcriptomics and proteomics, this study identifies a selective loss of PRR4⁺CST3⁺WFDC2⁻ seromucous acinar cells and a rise in GZMK⁺CD8⁺ T cells in Sjögren's disease, revealing distinct immune-mediated epithelial remodeling and interferon-driven dysfunction across diverse clinical presentations. These findings uncover a novel sub-cytolytic effector mechanism by which GZMK⁺CD8⁺ T cells impair mitochondrial integrity and activate innate immune signaling, linking epithelial injury to type I interferon responses and offering new therapeutic targets.

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Conflict of interest statement

Competing interests BMW reports a relationship with Pfizer Inc that includes nonfinancial support. BMW reports a relationship with Mitobridge Inc that includes nonfinancial support. SAT reports a relationship with Sanofi that includes board membership and consulting or advisory. SAT reports a relationship with GlaxoSmithKline Inc that includes board membership, consulting or advisory, and employment. SAT reports a relationship with Foresight Labs that includes board membership and consulting or advisory. SAT reports a relationship with QIAGEN GmbH that includes board membership and consulting or advisory. SAT reports a relationship with Xaira Therapeutics that includes board membership and consulting or advisory. SAT reports a relationship with Ensocell that includes equity or stocks. SAT reports a relationship with Transition Bio, Inc. that includes equity or stocks. SAT, KMB, QTE, BFM, and BMW, are all active members of the Human Cell Atlas. NIAMS has CRADAs with AstraZeneca and Bristol Myers Squibb. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Update of

GZMK+CD8+ T cells Target A Specific Acinar Cell Type in Sjögren's Disease.
Pranzatelli TJF, Perez P, Ku A, Matuck B, Huynh K, Sakai S, Abed M, Jang SI, Yamada E, Dominick K, Ahmed Z, Oliver A, Wasikowski R, Easter QT, Baer AN, Pelayo E, Khavandgar Z, Kleiner DE, Magone MT, Gupta S, Lessard C, Farris AD, Burbelo PD, Martin D, Morell RJ, Zheng C, Rachmaninoff N, Maldonado-Ortiz J, Qu X, Aure M, Dezfulian MH, Lake R, Teichmann S, Barber DL, Tsoi LC, Sowalsky AG, Tyc KM, Liu J, Gudjonsson J, Byrd KM, Johnson PLF, Chiorini JA, Warner BM. Pranzatelli TJF, et al. Res Sq [Preprint]. 2024 Jul 11:rs.3.rs-3601404. doi: 10.21203/rs.3.rs-3601404/v2. Res Sq. 2024. Update in: Ann Rheum Dis. 2025 Oct 28:S0003-4967(25)04313-4. doi: 10.1016/j.ard.2025.08.014. PMID: 38196575 Free PMC article. Updated. Preprint.

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GZMK⁺CD8⁺ T cells target a specific acinar cell type in Sjögren's disease

Affiliations

GZMK⁺CD8⁺ T cells target a specific acinar cell type in Sjögren's disease

Authors

Affiliations

Abstract

Conflict of interest statement

Update of

References

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