Lipid-Oriented Live-Cell Distinction of B and T Lymphocytes

Kwon, Hyeokjin; Das, Raj Kumar; Jung, Gun Tae; Lee, Hong-Guen; Lee, Sun Hyeok; Berry, Stuart N.; Tan, Justin Kok Soon; Park, Solip; Yang, Jun; Park, Soo-Hyun; Baek, Kangkyun; Park, Kyeng Min; Lee, Jae Won; Choi, Yunkyu; Kim, Ki Hean; Kim, Sangho; Kim, Kwang Pyo; Kang, Nam‐Young; Kim, Kimoon; Chang, Young‐Tae

doi:10.1021/jacs.1c00944

Cited by 24 publications

(22 citation statements)

References 28 publications

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“…This is a clear contrast to another B cell probe CDgB, whose mechanism is LOLD, based on the cell membrane flexibility. [6] The membrane flexibility decreases with the increasing chain length of fatty acid and cholesterol contents in the cell membrane. CDgB showed the brightest staining in immature B cells due to the softest membrane structure, and medium in matured B cells.…”

Section: Methodsmentioning

confidence: 99%

“…DOFL approach could not only offer a useful molecular probe, but also provide a chance to discover a novel biomarker for the target cell. Based on the DOFL approach, the first B‐cell‐selective probe CDgB over T cells, was recently reported with Lipid‐Oriented Live‐cell Distinction (LOLD) mechanism [6] . LOLD utilizes the small difference in membrane components and flexibility for cell discrimination [7] .…”

Section: Introductionmentioning

confidence: 99%

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A SLC35C2 Transporter‐Targeting Fluorescent Probe for the Selective Detection of B Lymphocytes Identified by SLC‐CRISPRi and Unbiased Fluorescence Library Screening

et al. 2022

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T and B lymphocytes are two major adaptive immune cells in the human defense system. To real-time monitor their diverse functions, a live-cell-selective probe for only one cell type is need to investigate the complex interaction of the immune cells. Herein, a small-molecule probe CDyB for live B cells is developed by an unbiased fluorescence library screening. The cell selectivity was confirmed by multiparametric single-cell analysis using CyTOF. Through a systematic SLC-CRISPRi library screening, the molecular target of CDyB was identified as SLC35C2 transporter based on a gating-oriented live-cell distinction (GOLD) mechanism. The gene expression analysis and knock-out experiments validated that the SLC35C2 transporter was the target for CDyB distinction. Interestingly, when CDyB was applied to study B cell development, the CDyB fluorescence and SLC35C2 expression were positively correlated with the B cell maturation process, and not involved in the T cell development.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A SLC35C2 Transporter‐Targeting Fluorescent Probe for the Selective Detection of B Lymphocytes Identified by SLC‐CRISPRi and Unbiased Fluorescence Library Screening

et al. 2022

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“…In a related approach, Kwon et al applied a combinatorial lipid-oriented library to distinguish T cells from B cells and identified a B cell selective probe (CDgB). 43 Notably, the authors used CDgB to selectively label B cells in mixtures of splenocytes without the help of antibodies. In another example, fluorescently labeled antibodies were implemented in FRET imaging studies to investigate the cell surface organization of interleukin-9 and interleukin-2 receptors (IL-9R and IL-2R, respectively), which have a crucial regulatory role in T cells.…”

Section: Probes For Imaging T Cells and Nk Cellsmentioning

confidence: 99%

Activatable Fluorophores for Imaging Immune Cell Function

Mendive‐Tapia

Vendrell

2022

Acc. Chem. Res.

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Conspectus Optical imaging has become an essential tool to study biomolecular processes in live systems with unprecedented spatial resolution. New fluorescent technologies and advances in optical microscopy have revolutionized the ways in which we can study immune cells in real time. For example, activatable fluorophores that emit signals after target recognition have enabled direct imaging of immune cell function with enhanced readouts and minimal background. In this Account, we summarize recent advances in the chemical synthesis and implementation of activatable fluorescent probes to monitor the activity and the role of immune cells in different pathological processes, from infection to inflammatory diseases or cancer. In addition to the contributions that our group has made to this field, we review the most relevant literature disclosed over the past decade, providing examples of different activatable architectures and their application in diagnostics and drug discovery. This Account covers the imaging of the three major cell types in the immune system, that is, neutrophils, macrophages, and lymphocytes. Attracted by the tunability and target specificity of peptides, many groups have designed strategies based on fluorogenic peptides whose fluorescence emission is regulated by the reaction with enzymes (e.g., MMPs, cathepsins, granzymes), or through Förster resonance energy transfer (FRET) mechanisms. Selective imaging of immune cells has been also achieved by targeting different intracellular metabolic routes, such as lipid biogenesis. Other approaches involve the implementation of diversity-oriented fluorescence libraries or the use of environmentally sensitive fluorescent scaffolds (e.g., molecular rotors). Our group has made important progress by constructing probes to image metastasis-associated macrophages in tumors, apoptotic neutrophils, or cytotoxic natural killer (NK) cells against cancer cells, among other examples. The chemical probes covered in this Account have been successfully validated in vitro in cell culture systems, and in vivo in relevant models of inflammation and cancer. Overall, the range of chemical structures and activation mechanisms reported to sense immune cell function is remarkable. However, the emergence of new strategies based on new molecular targets or activatable mechanisms that are yet to be discovered will open the door to track unexplored roles of immune cells in different biological systems. We anticipate that upcoming generations of activatable probes will find applications in the clinic to help assessing immunotherapies and advance precision medicine. We hope that this Account will evoke new ideas and innovative work in the design of fluorescent probes for imaging cell function.

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“…Emerging small molecule-based fluorescent probes have the potential to replace the antibodies. Recently, two fluorescent probes, CDgB and CDyB, for B cells’ distinction from T cells, have been developed by our group using unbiased screening with the Diversity-Oriented Fluorescence Library (DOFL), breaking the conventional method ( Figure 1 ) [ 10 , 11 ]. DOFL is a toolbox for cell-type distinction by combinatorial screening [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Human-T-Cell-Selective Fluorescent Probe

Gao

Chang

2022

Cells

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The identification of T and B lymphocytes has relied on using antibodies against different biomarkers as the gold standard. Emerging small molecule-based fluorescent probes have the potential to replace antibodies. Herein, we report the first human-T-cell-selective fluorescent probe, Mito thermo yellow (MTY), achieving the live T cells’ distinction from B cells, which was previously impossible without the help of antibodies. The unexpected cell selectivity of MTY is attributed to the higher mitochondria mass and membrane potential of T cells over B cells. This study enriches the toolbox for live cell distinction from complex cell communities.

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Lipid-Oriented Live-Cell Distinction of B and T Lymphocytes

Cited by 24 publications

References 28 publications

A SLC35C2 Transporter‐Targeting Fluorescent Probe for the Selective Detection of B Lymphocytes Identified by SLC‐CRISPRi and Unbiased Fluorescence Library Screening

A SLC35C2 Transporter‐Targeting Fluorescent Probe for the Selective Detection of B Lymphocytes Identified by SLC‐CRISPRi and Unbiased Fluorescence Library Screening

Activatable Fluorophores for Imaging Immune Cell Function

Human-T-Cell-Selective Fluorescent Probe

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