Mass cytometry staining for human bone marrow clinical samples

Hallisey, Margaret; Dennis, Jenna; Abrecht, Charlotte; Pistofidis, Romanos Sklavenitis; Schork, Abigail N.; Lightbody, Elizabeth D.; Heilpern-Mallory, Daniel; Severgnini, Mariano; Ghobrial, Irène M.; Hodi, F. Stephen; Bagińska, Joanna

doi:10.1016/j.xpro.2022.101163

“…The following morning, cells were washed with Maxpar® Cell Staining Buffer followed by Maxpar® Cell Acquisition Solution (CAS), filtered through a 35μm cell strainer and resuspended in Maxpar® CAS containing 0.1× EQ Four Element Calibration Beads at a cell concentration of 1 × 10 6 cells/ml. Cells were run on the Helios (Standard BioTools) instrument at Longwood Medical Area CyTOF Core at Dana-Farber Cancer Institute (DFCI) according to the standardized good clinical laboratory practice procedures 90 , 91 .…”

Section: Methodsmentioning

confidence: 99%

Immune profiling of age and adjuvant-specific activation of human blood mononuclear cells in vitro

Schüller,

Barman,

Mendez-Giraldez

et al. 2024

Commun Biol

2

0

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Vaccination reduces morbidity and mortality due to infections, but efficacy may be limited due to distinct immunogenicity at the extremes of age. This raises the possibility of employing adjuvants to enhance immunogenicity and protection. Early IFNγ production is a hallmark of effective vaccine immunogenicity in adults serving as a biomarker that may predict effective adjuvanticity. We utilized mass cytometry (CyTOF) to dissect the source of adjuvant-induced cytokine production in human blood mononuclear cells (BMCs) from newborns (~39-week-gestation), adults (~18-63 years old) and elders (>65 years of age) after stimulation with pattern recognition receptors agonist (PRRa) adjuvants. Dimensionality reduction analysis of CyTOF data mapped the BMC compartment, elucidated age-specific immune responses and profiled PRR-mediated activation of monocytes and DCs upon adjuvant stimulation. Furthermore, we demonstrated PRRa adjuvants mediated innate IFNγ induction and mapped NK cells as the key source of TLR7/8 agonist (TLR7/8a) specific innate IFNγ responses. Hierarchical clustering analysis revealed age and TLR7/8a-specific accumulation of innate IFNγ producing γδ T cells. Our study demonstrates the application of mass cytometry and cutting-edge computational approaches to characterize immune responses across immunologically distinct age groups and may inform identification of the bespoke adjuvantation systems tailored to enhance immunity in distinct vulnerable populations.

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“…The following morning, cells were washed with Maxpar® Cell Staining Buffer followed by Maxpar® Cell Acquisition Solution (CAS), filtered through a 35μm cell strainer and resuspended in Maxpar® CAS containing 0.1× EQ Four Element Calibration Beads at a cell concentration of 1 × 10 6 cells/ml. Cells were run on the Helios (Standard BioTools) instrument at Longwood Medical Area CyTOF Core at Dana-Farber Cancer Institute (DFCI) according to the standardized good clinical laboratory practice procedures 85,86 .…”

Section: Mass Cytometrymentioning

confidence: 99%

Immune profiling of age and adjuvant-specific activation of human blood mononuclear cells in vitro

Dowling,

Schüller,

Barman

et al. 2023

Preprint

1

0

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Vaccination reduces morbidity and mortality due to infections, but efficacy may be limited due to distinct immunogenicity at the extremes of age. This raises the possibility of employing adjuvants to enhance immunogenicity and protection. Early IFNγ production is a hallmark of effective vaccine immunogenicity in adults serving as a biomarker that may predict effective adjuvanticity. We utilized mass cytometry (CyTOF) to dissect the source of adjuvant-induced cytokine production in human blood mononuclear cells (BMCs) from newborns (~39-week-gestation), adults (~18-63 years old) and elders (>65 years of age) after stimulation with pattern recognition receptors agonist (PRRa) adjuvants. Dimensionality reduction analysis of CyTOF data mapped the BMC compartment, elucidated age-specific immune responses and profiled PRR-mediated activation of monocytes and DCs upon adjuvant stimulation. Furthermore, we demonstrated PRRa adjuvants mediated innate IFNγ induction and mapped NK cells as the key source of TLR7/8 agonist (TLR7/8a) specific innate IFNγ responses. Hierarchical clustering analysis revealed age and TLR7/8a-specific accumulation of innate IFNγ producing γδ T cells. Our study demonstrates the application of mass cytometry and cutting-edge computational approaches to characterize immune responses across immunologically distinct age groups and may inform identification of the bespoke adjuvantation systems tailored to enhance immunity in distinct vulnerable populations.

show abstract