Flow Cytometry: The Next Revolution DOI Creative Commons
J. Paul Robinson, Raluca Ostafe, Sharath Narayana Iyengar

et al.

Cells, Journal Year: 2023, Volume and Issue: 12(14), P. 1875 - 1875

Published: July 17, 2023

Unmasking the subtleties of immune system requires both a comprehensive knowledge base and ability to interrogate that with intimate sensitivity. That task, considerable extent, has been handled by an iterative expansion in flow cytometry methods, technological capability also accompanying advances informatics. As field fluorescence-based cytomics matured, it reached barrier at around 30 parameter analyses, which stalled until spectral created fundamental transformation will likely lead potential 100 simultaneous analyses within few years. The advance informatics now become watershed moment for as competes mature systematic approaches such genomics proteomics, allowing take seat multi-omics table. In addition, recent try combine speed systems other detection addition fluorescence alone, make flow-based instruments even more indispensable any biological laboratory. This paper outlines current cell analysis discusses traditional microfluidic sorting well next-generation instruments, provides early look future opportunities are arise.

Language: Английский

Marked T cell activation, senescence, exhaustion and skewing towards TH17 in patients with COVID-19 pneumonia DOI Creative Commons
Sara De Biasi, Marianna Meschiari, Lara Gibellini

et al.

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: July 6, 2020

Abstract The immune system of patients infected by SARS-CoV-2 is severely impaired. Detailed investigation T cells and cytokine production in affected COVID-19 pneumonia are urgently required. Here we show that, compared with healthy controls, patients’ cell compartment displays several alterations involving naïve, central memory, effector memory terminally differentiated cells, as well regulatory PD1 + CD57 exhausted cells. Significant exist also lineage-specifying transcription factors chemokine receptors. Terminally from proliferate less than those whereas their mitochondria functionality similar CD4 both groups. Patients display significant increases proinflammatory or anti-inflammatory cytokines, including helper type-1 type-2 chemokines galectins; lymphocytes produce more tumor necrosis factor (TNF), interferon-γ, interleukin (IL)-2 IL-17, the last observation implying that blocking IL-17 could provide a novel therapeutic strategy for COVID-19.

Language: Английский

Citations

730
Impaired immune cell cytotoxicity in severe COVID-19 is IL-6 dependent DOI Open Access
Alessio Mazzoni, Lorenzo Salvati, Laura Maggi

et al.

Journal of Clinical Investigation, Journal Year: 2020, Volume and Issue: 130(9), P. 4694 - 4703

Published: May 28, 2020

BACKGROUND. Coronavirus disease 19 (COVID-19) is an emerging infectious caused by SARS-CoV-2. Antiviral immune response crucial to achieve pathogen clearance; however, in some patients excessive and aberrant host can lead acute respiratory distress syndrome. The comprehension of the mechanisms that regulate elimination, immunity, pathology essential better characterize progression widen spectrum therapeutic options.

Language: Английский

Citations

503
Directed Evolution: Methodologies and Applications DOI
Yajie Wang, Pu Xue, Mingfeng Cao

et al.

Chemical Reviews, Journal Year: 2021, Volume and Issue: 121(20), P. 12384 - 12444

Published: July 23, 2021

Directed evolution aims to expedite the natural process of biological molecules and systems in a test tube through iterative rounds gene diversifications library screening/selection. It has become one most powerful widespread tools for engineering improved or novel functions proteins, metabolic pathways, even whole genomes. This review describes commonly used diversification strategies, screening/selection methods, recently developed continuous strategies directed evolution. Moreover, we highlight some representative applications nucleic acids, genetic circuits, viruses, cells. Finally, discuss challenges future perspectives

Language: Английский

Citations

450
Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) DOI Creative Commons
Andrea Cossarizza, Hyun‐Dong Chang, Andreas Radbruch

et al.

European Journal of Immunology, Journal Year: 2021, Volume and Issue: 51(12), P. 2708 - 3145

Published: Dec. 1, 2021

Abstract The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes functional assays all major human murine immune cell subsets. Notably, contain helpful tables highlighting differences between cells. Another useful feature this is analysis clinical samples with examples applications in context autoimmune diseases, cancers as well acute chronic infectious diseases. Furthermore, there are detailing tips, tricks pitfalls avoid. All written peer‐reviewed by leading experts immunologists, making an essential state‐of‐the‐art handbook for basic researchers.

Language: Английский

Citations

315
Meta-Analysis of Leukocyte Diversity in Atherosclerotic Mouse Aortas DOI Open Access
Alma Zernecke, Holger Winkels, Clément Cochain

et al.

Circulation Research, Journal Year: 2020, Volume and Issue: 127(3), P. 402 - 426

Published: July 16, 2020

The diverse leukocyte infiltrate in atherosclerotic mouse aortas was recently analyzed 9 single-cell RNA sequencing and 2 mass cytometry studies. In a comprehensive meta-analysis, we confirm 4 known macrophage subsets-resident, inflammatory, interferon-inducible cell, Trem2 (triggering receptor expressed on myeloid cells-2) foamy macrophages-and identify new subset resembling cavity macrophages. We also find that monocytes, neutrophils, dendritic cells, natural killer innate lymphoid cells-2, CD (cluster of differentiation)-8 T cells form prominent separate immune cell populations aortas. Many CD4 express IL (interleukin)-17 the chemokine CXCR (C-X-C receptor)-6. A small number regulatory helper 1 is identified. Immature naive are present both healthy Our meta-analysis overcomes limitations individual studies that, because their experimental approach, over- or underrepresent certain populations. Mass demonstrate surface phenotype provides valuable information beyond transcriptomes. analysis helps resolve some long-standing controversies field. First,

Language: Английский

Citations

296
Treg-expressed CTLA-4 depletes CD80/CD86 by trogocytosis, releasing free PD-L1 on antigen-presenting cells DOI Open Access
Murat Tekgüç, James B. Wing, Motonao Osaki

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2021, Volume and Issue: 118(30)

Published: July 23, 2021

Foxp3-expressing CD4+CD25+ regulatory T cells (Tregs) constitutively and highly express the immune checkpoint receptor cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4), whose Treg-specific deficiency causes severe systemic autoimmunity. As a key mechanism of Treg-mediated suppression, Treg-expressed CTLA-4 down-regulates expression CD80/CD86 costimulatory molecules on antigen-presenting (APCs). Here, we show that facilitated Treg-APC conjugation synapse formation. The synapses thus formed provided stable platform whereby Tregs were able to deplete APCs by extracting them via CTLA-4-dependent trogocytosis. depletion occurred even with solely expressing mutant form lacking cytoplasmic portion required for its endocytosis. trogocytosis also accelerated in vitro vivo passive transfer other membrane proteins lipid from without their significant reduction APC surface. Furthermore, CD80 down-regulation or blockade soluble CTLA-4-immunoglobulin (CTLA-4-Ig), respectively, disrupted cis-CD80/programmed death ligand-1 (PD-L1) heterodimers increased free PD-L1 dendritic (DCs), expanding phenotypically distinct population CD80lo PD-L1hi DCs. Thus, are inhibit cell stimulatory activity reducing This is exert dual suppressive effects responses limiting costimulation naïve increasing available inhibition programmed death-1 (PD-1)-expressing effector cells. Blockade PD-1/PD-L1 combination may therefore synergistically hinder thereby effectively enhancing responses, including tumor immunity.

Language: Английский

Citations

270
Microbiota-Induced Type I Interferons Instruct a Poised Basal State of Dendritic Cells DOI Creative Commons
Laura Schaupp, Sabine Muth,

Leif Rogell

et al.

Cell, Journal Year: 2020, Volume and Issue: 181(5), P. 1080 - 1096.e19

Published: May 1, 2020

Language: Английский

Citations

204
Untimely TGFβ responses in COVID-19 limit antiviral functions of NK cells DOI Creative Commons
Mario Witkowski, Caroline Tizian, Marta Ferreira‐Gomes

et al.

Nature, Journal Year: 2021, Volume and Issue: 600(7888), P. 295 - 301

Published: Oct. 25, 2021

Language: Английский

Citations

201
Precursors for Nonlymphoid-Tissue Treg Cells Reside in Secondary Lymphoid Organs and Are Programmed by the Transcription Factor BATF DOI Creative Commons
Michael Delacher, Charles D. Imbusch, Agnes Hotz‐Wagenblatt

et al.

Immunity, Journal Year: 2020, Volume and Issue: 52(2), P. 295 - 312.e11

Published: Jan. 7, 2020

Specialized regulatory T (Treg) cells accumulate and perform homeostatic regenerative functions in nonlymphoid tissues. Whether common precursors for nonlymphoid-tissue Treg exist how they differentiate remain elusive. Using transcription factor nuclear factor, interleukin 3 regulated (Nfil3) reporter mice single-cell RNA-sequencing (scRNA-seq), we identified two precursor stages of 33 (IL-33) receptor ST2-expressing tissue cells, which resided the spleen lymph nodes. Global chromatin profiling revealed a stepwise acquisition accessibility reprogramming toward cell phenotype. Mechanistically, validated Batf as driver molecular program precursors. Understanding this development will help to harness properties therapy.

Language: Английский

Citations

192
Discrete populations of isotype-switched memory B lymphocytes are maintained in murine spleen and bone marrow DOI Creative Commons
René Riedel, Richard Addo, Marta Ferreira‐Gomes

et al.

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: May 22, 2020

Abstract At present, it is not clear how memory B lymphocytes are maintained over time, and whether only as circulating cells or also residing in particular tissues. Here we describe distinct populations of isotype-switched (Bsm) murine spleen bone marrow, identified according to individual transcriptional signature cell receptor repertoire. A population marginal zone-like located exclusively the spleen, while a quiescent Bsm found marrow. Three further resident populations, present represent transitional follicular B1 cells, respectively. representing 10-20% marrow one qualifying circulating. In all individually dock onto VCAM1 + stromal and, reminiscent T plasma void activation, proliferation mobility.

Language: Английский

Citations

188