<scp>OMIP</scp>‐070: <scp>NKp46</scp>‐Based 27‐Color Phenotyping to Define Natural Killer Cells Isolated From Human Tumor Tissues

Frutoso, Marie; Mair, Florian; Prlic, Martin

doi:10.1002/cyto.a.24230

Cited by 7 publications

(5 citation statements)

References 54 publications

(68 reference statements)

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“…More CCL20 and CXCL16 was detected in periodontitis gingival tissue compared to healthy tissue which may act to recruit MAIT cells via CCR6 or CXCR6, respectively 98 . It is important to note that a comparison between blood and tissue derived MAIT cells needs to be done carefully and take any collagenase‐mediated effects on protein and transcript expression into account 99,100 . Further insight about potential changes is needed from studies that examine MAIT cells at the single‐cell level in the same tissue, but across a gradient of mild to severe inflammation (which is feasible with gingival tissues 101 ), which would allow for the assessment of inflammation‐associated MAIT cell activation states.…”

Section: Mait Cell Phenotype and Function In A Sterile Versus Bacteri...mentioning

confidence: 99%

Signals that control MAIT cell function in healthy and inflamed human tissues

Konecny,

Huang,

Setty

et al. 2024

Immunological Reviews

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SummaryMucosal‐associated invariant T (MAIT) cells have a semi‐invariant T‐cell receptor that allows recognition of antigen in the context of the MHC class I‐related (MR1) protein. Metabolic intermediates of the riboflavin synthesis pathway have been identified as MR1‐restricted antigens with agonist properties. As riboflavin synthesis occurs in many bacterial species, but not human cells, it has been proposed that the main purpose of MAIT cells is antibacterial surveillance and protection. The majority of human MAIT cells secrete interferon‐gamma (IFNg) upon activation, while some MAIT cells in tissues can also express IL‐17. Given that MAIT cells are present in human barrier tissues colonized by a microbiome, MAIT cells must somehow be able to distinguish colonization from infection to ensure effector functions are only elicited when necessary. Importantly, MAIT cells have additional functional properties, including the potential to contribute to restoring tissue homeostasis by expression of CTLA‐4 and secretion of the cytokine IL‐22. A recent study provided compelling data indicating that the range of human MAIT cell functional properties is explained by plasticity rather than distinct lineages. This further underscores the necessity to better understand how different signals regulate MAIT cell function. In this review, we highlight what is known in regards to activating and inhibitory signals for MAIT cells with a specific focus on signals relevant to healthy and inflamed tissues. We consider the quantity, quality, and the temporal order of these signals on MAIT cell function and discuss the current limitations of computational tools to extrapolate which signals are received by MAIT cells in human tissues. Using lessons learned from conventional CD8 T cells, we also discuss how TCR signals may integrate with cytokine signals in MAIT cells to elicit distinct functional states.

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Section: Mait Cell Phenotype and Function In A Sterile Versus Bacteri...mentioning

confidence: 99%

Signals that control MAIT cell function in healthy and inflamed human tissues

Konecny,

Huang,

Setty

et al. 2024

Immunological Reviews

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“…Thus, sample type should be selected based on the questions being asked. Of note, the collagenase digestion required to extract immune cells from tissues affects the transcriptional profile of T cells 78 and may also affect the detection of some surface proteins by antibodies, 79 and this must be considered and carefully assessed when using biopsies. Similarly, proper study design and analysis methods are required to avoid introduction of batch effects in studies 80 .…”

Section: How To Assess Trm Cells In the Human Frt—common Sampling Met...mentioning

confidence: 99%

Advances and challenges in studying the tissue‐resident T cell compartment in the human female reproductive tract

Lund

Hladik

Prlic

2023

Immunological Reviews

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Summary Tissue‐resident memory T cells (TRM) are considered to be central to maintaining mucosal barrier immunity and tissue homeostasis. Most of this knowledge stems from murine studies, which provide access to all organs. These studies also allow for a thorough assessment of the TRM compartment for each tissue and across tissues with well‐defined experimental and environmental variables. Assessing the functional characteristics of the human TRM compartment is substantially more difficult; thus, notably, there is a paucity of studies profiling the TRM compartment in the human female reproductive tract (FRT). The FRT is a mucosal barrier tissue that is naturally exposed to a wide range of commensal and pathogenic microbes, including several sexually transmitted infections of global health significance. We provide an overview of studies describing T cells within the lower FRT tissues and highlight the challenges of studying TRM cells in the FRT: different sampling methods of the FRT greatly affect immune cell recovery, especially of TRM cells. Furthermore, menstrual cycle, menopause, and pregnancy affect FRT immunity, but little is known about changes in the TRM compartment. Finally, we discuss the potential functional plasticity of the TRM compartment during inflammatory episodes in the human FRT to maintain protection and tissue homeostasis, which are required to ensure reproductive fitness.

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“…In short, single stain controls were made for Qdot 605 :: CD2 where 1 mM EDTA final concentration was added to each buffer during and after the staining process or not. The MFI of Qdot 605 was compared between the two conditions to determine whether quenching was taking place due to contamination (16). Only non-contaminated reagents were used without the addition of EDTA going forward (data not shown).…”

Section: Online Materialsmentioning

confidence: 99%

50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells

Konecny,

Mage,

Tyznik

et al. 2023

Preprint

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We report the development of an optimized 50-color spectral flow cytometry panel designed for the in-depth analysis of the immune system in human blood and tissues, with the goal of maximizing the amount of information that can be collected using currently available flow cytometry platforms. We established and tested this panel using peripheral blood mononuclear cells (PBMCs), but included CD45 to enable its use for the analysis of human tissue samples. The panel contains lineage markers for all major immune cell subsets, and an extensive set of phenotyping markers focused on the activation and differentiation status of the T cell and dendritic cell (DC) compartment.We outline the biological insight that can be gained from the simultaneous measurement of such a large number of proteins and propose that this approach provides a unique opportunity for the comprehensive exploration of the immune status in tissue biopsies and other human samples with a limited number of cells. Of note, we tested the panel to be compatible with cell sorting for further downstream applications. Furthermore, to facilitate the wide-spread implementation of such a panel across different cohorts and samples, we established a trimmed-down 45-color version which can be used with different spectral cytometry platforms.Finally, to generate this panel, we utilized not only existing panel design guidelines, but also developed new metrics to systematically identify the optimal combination of 50 fluorochromes and evaluate fluorochrome-specific resolution in the context of a 50-color unmixing matrix.

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OMIP‐070: NKp46‐Based 27‐Color Phenotyping to Define Natural Killer Cells Isolated From Human Tumor Tissues

Cited by 7 publications

References 54 publications

Signals that control MAIT cell function in healthy and inflamed human tissues

Signals that control MAIT cell function in healthy and inflamed human tissues

Advances and challenges in studying the tissue‐resident T cell compartment in the human female reproductive tract

50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells

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