Standardization and quality control for high-dimensional mass cytometry studies of human samples

Katja Kleinsteuber; Björn Corleis; Narges Rashidi; Nzuekoh Nchinda; Antonella Lisanti; Josalyn L Cho; Benjamin D Medoff; Douglas Kwon; Bruce D Walker

doi:10.1002/cyto.a.22935

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Standardization and quality control for high-dimensional mass cytometry studies of human samples

Journal article

Peer reviewed

Standardization and quality control for high-dimensional mass cytometry studies of human samples

Katja Kleinsteuber, Björn Corleis, Narges Rashidi, Nzuekoh Nchinda, Antonella Lisanti, Josalyn L Cho, Benjamin D Medoff, Douglas Kwon and Bruce D Walker

Cytometry. Part A, Vol.89(10), pp.903-913

10/2016

DOI: 10.1002/cyto.a.22935

PMCID: PMC5495108

PMID: 27575385

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Abstract

Mass cytometry (CyTOF), a mass spectrometry-based single cell phenotyping technology, allows utilization of over 35 antibodies in a single sample and is a promising tool for translational human immunology studies. Although several analysis tools are available to interpret the complex data sets generated, a robust method for standardization and quality control within and across studies is needed. Here we report an efficient and easily adaptable method to monitor quality of individual samples in human immunology studies and to facilitate reproducible data analysis. Samples to be assessed are spiked with a defined amount of reference peripheral blood mononuclear cells from a healthy donor, derived from a single large blood draw. The presence of known standardized numbers and phenotypic profiles of these reference cells greatly facilitates sample analysis by allowing for: 1) quality control for consistent staining of each antibody in the panel, 2) identification of potential batch effects, and 3) implementation of a robust gating strategy. We demonstrate the utility of this method using peripheral blood and bronchoalveolar lavage samples from HIV patients by characterizing their CD8 T-cell phenotypes and cytokine expression, respectively. Our results indicate that this method allows quality control of experimental conditions and results in highly reproducible population frequencies through a robust gating strategy. © 2016 International Society for Advancement of Cytometry.

CD8-Positive T-Lymphocytes - cytology

Leukocytes, Mononuclear - metabolism

Flow Cytometry - methods

Reproducibility of Results

Antibodies - metabolism

Cytokines - metabolism

Humans

Bronchoalveolar Lavage - methods

Reference Standards

CD8-Positive T-Lymphocytes - metabolism

Immunophenotyping - methods

Leukocytes, Mononuclear - cytology

Mass Spectrometry - methods

HIV Infections - metabolism

Quality Control

Details

Title: Subtitle: Standardization and quality control for high-dimensional mass cytometry studies of human samples
Creators: Katja Kleinsteuber - Heinrich-Pette Institute for Experimental Virology, Hamburg, Germany
Björn Corleis - Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA
Narges Rashidi - Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA
Nzuekoh Nchinda - Howard Hughes Medical Institute (HHMI) Chevy Chase, Maryland, USA
Antonella Lisanti - Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA
Josalyn L Cho - Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA
Benjamin D Medoff - Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA
Douglas Kwon - Infectious Disease Division, Massachusetts General Hospital, Boston, Massachusetts, USA
Bruce D Walker - Infectious Disease Division, Massachusetts General Hospital, Boston, Massachusetts, USA. bwalker@mgh.harvard.edu
Resource Type: Journal article
Publication Details: Cytometry. Part A, Vol.89(10), pp.903-913
DOI: 10.1002/cyto.a.22935
PMID: 27575385
PMCID: PMC5495108
NLM abbreviation: Cytometry A
ISSN: 1552-4922
eISSN: 1552-4930
Grant note: UM1 AI100663 / NIAID NIH HHS P30 AI060354 / NIAID NIH HHS U01 HL121827 / NHLBI NIH HHS
Language: English
Date published: 10/2016
Academic Unit: Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
Record Identifier: 9984094328802771

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