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PLoS One. 2015 Mar 23;10(3):e0121546. doi: 10.1371/journal.pone.0121546. eCollection 2015.

A method for identification and analysis of non-overlapping myeloid immunophenotypes in humans.

Author information

1
Human Cellular Therapy Laboratory, Division of Transfusion Medicine, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States of America.
2
Division of Hematology, Department of Medicine, Mayo Clinic, Rochester, MN, United States of America.
3
Department of Immunology, Mayo Clinic, Rochester, MN, United States of America.
4
Pulmonary and Critical Care Medicine, Mayo Clinic, Rochester, MN, United States of America.
5
Human Cellular Therapy Laboratory, Division of Transfusion Medicine, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States of America; Division of Hematology, Department of Medicine, Mayo Clinic, Rochester, MN, United States of America.

Abstract

The development of flow cytometric biomarkers in human studies and clinical trials has been slowed by inconsistent sample processing, use of cell surface markers, and reporting of immunophenotypes. Additionally, the function(s) of distinct cell types as biomarkers cannot be accurately defined without the proper identification of homogeneous populations. As such, we developed a method for the identification and analysis of human leukocyte populations by the use of eight 10-color flow cytometric protocols in combination with novel software analyses. This method utilizes un-manipulated biological sample preparation that allows for the direct quantitation of leukocytes and non-overlapping immunophenotypes. We specifically designed myeloid protocols that enable us to define distinct phenotypes that include mature monocytes, granulocytes, circulating dendritic cells, immature myeloid cells, and myeloid derived suppressor cells (MDSCs). We also identified CD123 as an additional distinguishing marker for the phenotypic characterization of immature LIN-CD33+HLA-DR- MDSCs. Our approach permits the comprehensive analysis of all peripheral blood leukocytes and yields data that is highly amenable for standardization across inter-laboratory comparisons for human studies.

PMID:
25799053
PMCID:
PMC4370675
DOI:
10.1371/journal.pone.0121546
[Indexed for MEDLINE]
Free PMC Article

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