title: Standardization and quality control for high-dimensional mass cytometry studies of human samples.
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.
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute (HHMI) Chevy Chase, Maryland, USA.|Heinrich-Pette Institute for Experimental Virology, Hamburg, Germany.
authors: Katja
authors: K
authors: Kleinsteuber
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.
authors: Björn
authors: B
authors: Corleis
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.
authors: Narges
authors: N
authors: Rashidi
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute (HHMI) Chevy Chase, Maryland, USA.
authors: Nzuekoh
authors: N
authors: Nchinda
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.
authors: Antonella
authors: A
authors: Lisanti
authors: Division of Pulmonary and Critical Care Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.|Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
authors: Josalyn L
authors: JL
authors: Cho
authors: Division of Pulmonary and Critical Care Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.|Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
authors: Benjamin D
authors: BD
authors: Medoff
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.|Infectious Disease Division, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors: Douglas
authors: D
authors: Kwon
authors: Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA. bwalker@mgh.harvard.edu.|Howard Hughes Medical Institute (HHMI) Chevy Chase, Maryland, USA. bwalker@mgh.harvard.edu.|Infectious Disease Division, Massachusetts General Hospital, Boston, Massachusetts, USA. bwalker@mgh.harvard.edu.
authors: Bruce D
authors: BD
authors: Walker
mesh_terms: D000906:Antibodies / Q000378:metabolism; D018893:Bronchoalveolar Lavage / Q000379:methods; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D015658:HIV Infections / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D016207:Cytokines
keywords: CyTOF; HIV; Mass cytometry; clinical studies; human immunology