<scp>OMIP 076: High‐dimensional</scp> immunophenotyping of murine T‐cell, B‐cell, and antibody secreting cell subsets

Mincham, Kyle T; Young, Jacob D; Strickland, Deborah H.

doi:10.1002/cyto.a.24474

Cited by 9 publications

(9 citation statements)

References 44 publications

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“…In conclusion, we present a full spectrum, 40-color immunophenotyping panel for robust exploration of all major leukocyte populations across primary and secondary lymphoid tissues. While we have personally focused on cancer research, a panel of such complexity is not limited to a single application and will be extremely valuable for study- with the use of CD3ε, CD4, CD8α, CD44, and CD62L to phenotype T cell subsets in the spleen [130][131][132][133]. OMIP-054 uses CyTOF in an immunocompetent mouse model of glioblastoma, but shares some similarities in the broad immunophenotyping efforts, spleen and bone marrow tissues studied, and marker choice [134].…”

Section: Discussionmentioning

confidence: 99%

“…Our panel shares similarity to OMIPs −031, −032, −076, and − 079 with the use of CD3ε, CD4, CD8α, CD44, and CD62L to phenotype T cell subsets in the spleen [130–133]. OMIP‐054 uses CyTOF in an immunocompetent mouse model of glioblastoma, but shares some similarities in the broad immunophenotyping efforts, spleen and bone marrow tissues studied, and marker choice [134].…”

Section: Discussionmentioning

confidence: 99%

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OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

Kare,

Nichols,

Zermeno

et al. 2023

Cytometry Pt A

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High‐dimensional immunoprofiling is essential for studying host response to immunotherapy, infection, and disease in murine model systems. However, the difficulty of multiparameter panel design combined with a lack of existing murine tools has prevented the comprehensive study of all major leukocyte phenotypes in a single assay. Herein, we present a 40‐color flow cytometry panel for deep immunophenotyping of murine lymphoid tissues, including the spleen, blood, Peyer's patches, inguinal lymph nodes, bone marrow, and thymus. This panel uses a robust set of surface markers capable of differentiating leukocyte subsets without the use of intracellular staining, thus allowing for the use of cells in downstream functional experiments or multiomic analyses. Our panel classifies T cells, B cells, natural killer cells, innate lymphoid cells, monocytes, macrophages, dendritic cells, basophils, neutrophils, eosinophils, progenitors, and their functional subsets by using a series of co‐stimulatory, checkpoint, activation, migration, and maturation markers. This tool has a multitude of systems immunology applications ranging from serial monitoring of circulating blood signatures to complex endpoint analysis, especially in pre‐clinical settings where treatments can modulate leukocyte abundance and/or function. Ultimately, this 40‐color panel resolves a diverse array of immune cells on the axes of time, tissue, and treatment, filling the niche for a modern tool dedicated to murine immunophenotyping.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

Kare,

Nichols,

Zermeno

et al. 2023

Cytometry Pt A

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“…OMIP‐031 focuses on co‐inhibitory molecules on T cell effector and memory subsets, which are also part of this panel [54]. OMIP‐076 phenotypes distinct T and B cell populations [55]. OMIP‐079 was designed to identify T cell subsets and analyze their proliferation using Ki‐67 [56].…”

Section: Similarity To Other Omipsmentioning

confidence: 99%

OMIP‐93: A 41‐color high parameter panel to characterize various co‐inhibitory molecules and their ligands in the lymphoid and myeloid compartment in mice

et al. 2023

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This 41‐color panel has been designed to characterize both the lymphoid and the myeloid compartments in mice. The number of immune cells isolated from organs is often low, whilst an increasing number of factors need to be analyzed to gain a deeper understanding of the complexity of an immune response. With a focus on T cells, their activation and differentiation status, as well as their expression of several co‐inhibitory and effector molecules, this panel also allows the analysis of ligands to these co‐inhibitory molecules on antigen‐presenting cells. This panel enables deep phenotypic characterization of CD4+ and CD8+ T cells, regulatory T cells, γδ T cells, NK T cells, B cells, NK cells, monocytes, macrophages, dendritic cells, and neutrophils. Whilst previous panels have focused on these topics individually, this is the first panel to enable simultaneous analysis of these compartments, thus enabling a comprehensive analysis with a limited number of immune cells/sample size. This panel is designed to analyze and compare the immune response in different mouse models of infectious diseases, but can also be extended to other disease models, for example tumors or autoimmune diseases. Here, we apply this panel to C57BL/6 mice infected with Plasmodium berghei ANKA, a mouse model of cerebral malaria.

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“…This OMIP panel has similarities with OMIP-057, OMIP-076, OMIP-079 for characterization of T Cells [47][48][49]. The panel also overlaps with OMIP-031, which aims to characterize tumorinfiltrating T cells and the expression of immunologic checkpoint in the MC38 model [1].…”

Section: Similarity To Other Published Omipsmentioning

confidence: 99%

A 30-Color Full-Spectrum Flow Cytometry Panel to Characterize the Immune Cell Landscape in Spleen and Tumors within a Syngeneic MC-38 Murine Colon Carcinoma Model

DeNiro

Que

Koo

et al. 2023

Preprint

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This panel was designed to characterize the immune cell landscape in the mouse tumor micro-environment as well as mouse lymphoid tissues (e.g., spleen). Previous Optimized Multicolor Immunofluorescence Panels (OMIP) with conventional cytometry were examples of high-quality fluorescent-based flow cytometry panels to characterize either the T-cell compartment or the myeloid compartment [1, 2]. The advent of spectral cytometry has enabled sufficient markers to be included in a panel to comprehensively characterize the immune cell landscape including both the T-cell and the myeloid cell compartments. In this body of work, we demonstrated that we could measure the frequency and characterize the functional status of CD4 T cells, CD8 T cells, regulatory T cells, NK cells, B cells, macrophages, granulocytes, monocytes, & dendritic cells. This panel is especially useful for understanding the immune landscape in "cold" preclinical tumor models with very low immune cell infiltration, and for investigating how therapeutic treatments may modulate the immune landscape.

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OMIP 076: High‐dimensional immunophenotyping of murine T‐cell, B‐cell, and antibody secreting cell subsets

Cited by 9 publications

References 44 publications

OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

OMIP‐93: A 41‐color high parameter panel to characterize various co‐inhibitory molecules and their ligands in the lymphoid and myeloid compartment in mice

A 30-Color Full-Spectrum Flow Cytometry Panel to Characterize the Immune Cell Landscape in Spleen and Tumors within a Syngeneic MC-38 Murine Colon Carcinoma Model

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