Morphological profiling of human T and NK lymphocytes by high-content cell imaging

  • Cell Rep. 2021 Jul 6;36(1):109318. doi: 10.1016/j.celrep.2021.109318.
Yolla German  1 ,  Loan Vulliard  2 ,  Anton Kamnev  3 ,  Laurène Pfajfer  1 ,  Jakob Huemer  4 ,  Anna-Katharina Mautner  3 ,  Aude Rubio  5 ,  Artem Kalinichenko  4 ,  Kaan Boztug  6 ,  Audrey Ferrand  5 ,  Jörg Menche  7 ,  Loïc Dupré  8
Affiliations
  • 1. Toulouse Institute for Infectious and Inflammatory Diseases (INFINITy), INSERM UMR1291, CNRS UMR5051, Toulouse III Paul Sabatier University, Toulouse, France; Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria.
  • 2. CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Department of Structural and Computational Biology, Max Perutz Labs, University of Vienna, Vienna, Austria.
  • 3. Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria; Department of Dermatology, Medical University of Vienna, Vienna, Austria.
  • 4. Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria; St. Anna Children's Cancer Research Institute (CCRI), Vienna, Austria.
  • 5. IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, 31024 Toulouse, France.
  • 6. Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; St. Anna Children's Cancer Research Institute (CCRI), Vienna, Austria; Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Vienna, Austria; St. Anna Children's Hospital, Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Vienna, Austria.
  • 7. CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Department of Structural and Computational Biology, Max Perutz Labs, University of Vienna, Vienna, Austria; Faculty of Mathematics, University of Vienna, Vienna, Austria.
  • 8. Toulouse Institute for Infectious and Inflammatory Diseases (INFINITy), INSERM UMR1291, CNRS UMR5051, Toulouse III Paul Sabatier University, Toulouse, France; Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria; Department of Dermatology, Medical University of Vienna, Vienna, Austria. Electronic address: [email protected].
PMID: 34233185 DOI: 10.1016/j.celrep.2021.109318
Abstract

The immunological synapse is a complex structure that decodes stimulatory signals into adapted lymphocyte responses. It is a unique window to monitor lymphocyte activity because of development of systematic quantitative approaches. Here we demonstrate the applicability of high-content imaging to human T and natural killer (NK) cells and develop a pipeline for unbiased analysis of high-definition morphological profiles. Our approach reveals how distinct facets of actin Cytoskeleton remodeling shape immunological synapse architecture and affect lytic granule positioning. Morphological profiling of CD8+ T cells from immunodeficient individuals allows discrimination of the roles of the ARP2/3 subunit ARPC1B and the ARP2/3 activator Wiskott-Aldrich syndrome protein (WASP) in immunological synapse assembly. Single-cell analysis further identifies uncoupling of lytic granules and F-actin radial distribution in ARPC1B-deficient lymphocytes. Our study provides a foundation for development of morphological profiling as a scalable approach to monitor primary lymphocyte responsiveness and to identify complex aspects of lymphocyte micro-architecture.

Keywords
ARPC1B deficiency; NK cells; WASP deficiency; actin cytoskeleton; cytotoxic T lymphocytes; high-content imaging; immunological synapse; lytic granules; morphological profiling.
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