OMIP‐022: Comprehensive assessment of antigen‐specific human T‐cell functionality and memory

Graves, Andrew J.; Padilla, Marcelino; Hokey, David A.

doi:10.1002/cyto.a.22478

Cited by 18 publications

(18 citation statements)

References 32 publications

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“…Cryopreserved PBMC (5 donors) and CBMC (14 donors) were thawed and incubated overnight (18 h) in RPMI 1640 medium supplemented with 10% FBS at 37 • C, 5% CO 2 . The cells were then washed with buffered saline and stained with a viability marker (Fixable Aqua Dead Cell Stain Kit, Thermo Fisher Scientific, San Diego, CA) prior to surface and intracellular staining with fluorescently conjugated antibodies using standard techniques as described (31). The staining panel used to identify T cell subsets and phenotypes was based on the Optimized Multicolor Immunofluorescence Panel 22 (OMIP-22) described previously (31).…”

Section: Phenotypic Characterization Of T Cellsmentioning

confidence: 99%

“…The cells were then washed with buffered saline and stained with a viability marker (Fixable Aqua Dead Cell Stain Kit, Thermo Fisher Scientific, San Diego, CA) prior to surface and intracellular staining with fluorescently conjugated antibodies using standard techniques as described (31). The staining panel used to identify T cell subsets and phenotypes was based on the Optimized Multicolor Immunofluorescence Panel 22 (OMIP-22) described previously (31). Fluorescently conjugated Abs used for cell surface staining were: APC-eFluor780-CD4 (clone RPA-T4, ebioscience, San Diego, CA); Horizon V500-CD14 (clone M5E2, BD Biosciences, San Jose, CA); Horizon V500-CD19 (clone HIB19, BD Biosciences); BV785-CD45RO (clone UCHL1, Biolegend, San Diego, CA); BV605-CCR7 (clone G043H7, Biolegend); and APC-CD366 (clone F38-2E2, Biolegend).…”

Section: Phenotypic Characterization Of T Cellsmentioning

confidence: 99%

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Redirection of Cord Blood T Cells and Natural Killer Cells for Elimination of Autologous HIV-1-Infected Target Cells Using Bispecific DART® Molecules

et al. 2020

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Mother-to-child transmission of HIV-1 remains a major global health challenge. Currently, HIV-1-infected infants require strict lifelong adherence to antiretroviral therapy to prevent replication of virus from reservoirs of infected cells, and to halt progression of disease. There is a critical need for immune interventions that can be deployed shortly after infection to eliminate HIV-1-infected cells in order to promote long-term remission of viremia, or to potentially cure pediatric HIV-1-infection. Bispecific HIV × CD3 DART ® molecules able to co-engage the HIV-1 envelope protein on the surface of infected cells and CD3 on cytolytic T cells have been previously shown to eliminate HIV-1 infected cells in vitro and are candidates for passive immunotherapy to reduce the virus reservoir. However, their potential utility as therapy for infant HIV-1 infection is unclear as the ability of these novel antibody-based molecules to work in concert with cells of the infant immune system had not been assessed. Here, we use human umbilical cord blood as a model of the naïve neonatal immune system to evaluate the ability of HIV x CD3 DART molecules to recruit and redirect neonatal effector cells for elimination of autologous CD4 + T cells infected with HIV-1 encoding an envelope gene sequenced from a motherto-child transmission event. We found that HIV × CD3 DART molecules can redirect T cells present in cord blood for elimination of HIV-infected CD4 + T cells. However, we observed reduced killing by T cells isolated from cord blood when compared to cells isolated from adult peripheral blood-likely due to the absence of the memory and effector CD8 + T cells that are most cytolytic when redirected by bispecific DART molecules. We also found that newly developed HIV × CD16 DART molecules were able to recruit CD16-expressing natural killer cells from cord blood to eliminate HIV-infected cells, and the activity of cord blood natural killer cells could be substantially increased Pollara et al. DART-Molecule Recruitment of Neonatal Cells by priming with IL-15. Our results support continued development of HIV-specific DART molecules using relevant preclinical animal models to optimize strategies for effective use of this immune therapy to reduce HIV-1 infection in pediatric populations.

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Section: Phenotypic Characterization Of T Cellsmentioning

confidence: 99%

Section: Phenotypic Characterization Of T Cellsmentioning

confidence: 99%

Redirection of Cord Blood T Cells and Natural Killer Cells for Elimination of Autologous HIV-1-Infected Target Cells Using Bispecific DART® Molecules

et al. 2020

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“…This panel is an expansion and modification of the OMIP‐014 panel , which now includes differentiation and T FH markers, whereas MIP‐1β and CD107a were removed. In addition, it presents similarities to other OMIP panels with intracellular cytokine staining to identify antigen specific T cells: OMIP‐001 , OMIP‐008 , OMIP‐009 , and OMIP‐022 . However, our panel includes T FH markers, the NK marker CD56, and cytokine IL‐21, which are not addressed in any of the other ICS OMIP panels.…”

Section: Similarity To Published Omipsmentioning

confidence: 96%

OMIP‐025: Evaluation of human T‐ and NK‐cell responses including memory and follicular helper phenotype by intracellular cytokine staining

et al. 2014

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Purpose and Appropriate Sample Types This panel was developed to assess antigen-specific T cells using peptide pools to various antigens of interest, although other types of antigens such as recombinant proteins or whole pathogens could be considered using different stimulation times. In addition to multiple functional markers, the panel includes differentiation markers and markers to assess follicular helper T cells and NK cells (Table 1). It was optimized using cryopreserved peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV) uninfected and HIV infected adults with known cytomegalovirus (CMV) responses and it underwent assay qualification. The panel is being used to evaluate the responses to HIV and malaria vaccine candidates in adults and children from different geographic areas.

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“…There are currently 22 unique panels described in this publication format, to query, e.g. antigen‐specific cells , chemokine receptors , B cells , innate cells , regulatory T cells , and gamma‐delta T cells .…”

Section: The Miner's Toolbox: Available Single Cell Technologiesmentioning

confidence: 99%

A Mine Is a Terrible Thing to Waste: High Content, Single Cell Technologies for Comprehensive Immune Analysis

Chattopadhyay

Roederer

2015

American Journal of Transplantation

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In recent years, an incredible variety of single cell technologies have become available to analyze immune responses. These technologies include polychromatic flow cytometry, mass cytometry, highly multiplexed single cell qPCR, RNA sequencing, microtools, and high‐resolution imaging. In this article, we review these platforms, describing their power and limitations for comprehensive analysis of the immune system. We relate the properties of these technologies to the various cellular states relevant to an immune response, in order to address which technologies are most appropriate for which settings.

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OMIP‐022: Comprehensive assessment of antigen‐specific human T‐cell functionality and memory

Cited by 18 publications

References 32 publications

Redirection of Cord Blood T Cells and Natural Killer Cells for Elimination of Autologous HIV-1-Infected Target Cells Using Bispecific DART® Molecules

Redirection of Cord Blood T Cells and Natural Killer Cells for Elimination of Autologous HIV-1-Infected Target Cells Using Bispecific DART® Molecules

OMIP‐025: Evaluation of human T‐ and NK‐cell responses including memory and follicular helper phenotype by intracellular cytokine staining

A Mine Is a Terrible Thing to Waste: High Content, Single Cell Technologies for Comprehensive Immune Analysis

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