Multi-color flow cytometry is the standard approach for immunophenotyping clinical samples. With the recent advances in cancer immunotherapy, myeloid-derived suppressor cells (MDSC), immature myeloid-lineage cells in cancer patient blood and the tumor microenvironment, are highlighted as an important immune cell population that correlates with prognosis and therapeutic efficacy. In contrast to their clear functions and existence, immunophenotyping of MDSC is not consistent among investigators due to surface antigens overlapping with many normal hematopoietic lineage cell populations. We performed a clinical study and analyzed more than 1000 breast cancer patients blood samples to quantitate MDSC during breast cancer progression. In this methodology manuscript, we described detailed procedures for study design, sample logistics and handling, staining and flow cytometric analysis. This protocol used a 7-color fluorochrome-conjugated antibody panel to analyze polymorphonuclear (PMN)- and monocytic (M)-MDSC subsets simultaneously. The interim analysis results of this study showed that both PMN and M-MDSC populations are increased in patients with bone metastasis compared with patients with visceral organ metastasis. In conclusion, this work provides a versatile, comprehensive, and practical protocol to measure MDSC in patient blood samples.
Bibliographical noteFunding Information:
The Korea University Research Grant; National R&D Program for Cancer Control , the Ministry of Health and Welfare, the Republic of Korea ( HA17C0040 ); The BK21 Plus Program, Korea University College of Medicine; and the National Research Foundation of the Republic of Korea ( 2020R1F1A1076996 and 2020R1A2C1012966 ).
© 2022 Elsevier B.V.
- Breast cancer
- Flow cytometry
- Myeloid-derived suppressor cells
ASJC Scopus subject areas
- Immunology and Allergy