Sensitive intranuclear flow cytometric quantification of IRF4 protein in multiple myeloma and normal human hematopoietic cells

• Published in: STAR protocols Vol. 2; no. 2; p. 100565
• Main Authors: Vora, Ashni A., Mondala, Phoebe K., Costello, Caitlin, MacLeod, A. Robert, Crews, Leslie A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.06.2021; Elsevier
• Subjects: Bone Marrow Cells - metabolism; Cancer; Cell isolation; Flow Cytometry - methods; Flow Cytometry/Mass Cytometry; Humans; Immunology; Interferon Regulatory Factors - metabolism; Limit of Detection; Multiple Myeloma - metabolism; Multiple Myeloma - pathology; Protocol; Stem Cells; Stem Cells; Immunology; Cell isolation; Flow Cytometry/Mass Cytometry; Cancer
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100565

Interferon regulatory factor 4 (IRF4) is a transcription factor that regulates normal and malignant immune cell development and is implicated in multiple myeloma pathogenesis. This protocol describes the use of combined cell surface and intranuclear staining with fluorescent antibodies to measure IRF4 protein expression within myeloma and normal immune cells. IRF4 protein quantification may provide a valuable prognostic tool to predict disease severity and sensitivity to IRF4-targeted therapies. This flow-cytometry-based procedure could also be rapidly translated into a clinically compatible assay. For complete details on the use and execution of this protocol, please refer to Mondala et al. (2021). [Display omitted] •Protocol for the quantification of human IRF4 levels by multi-parameter flow cytometry•Facilitates sensitive detection of IRF4 protein in myeloma cells and normal bone marrow•High-risk myeloma is typified by a population of CD38-high, IRF4-enriched cells Interferon regulatory factor 4 (IRF4) is a transcription factor that regulates normal and malignant immune cell development and is implicated in multiple myeloma pathogenesis. This protocol describes the use of combined cell surface and intranuclear staining with fluorescent antibodies to measure IRF4 protein expression within myeloma and normal immune cells. IRF4 protein quantification may provide a valuable prognostic tool to predict disease severity and sensitivity to IRF4-targeted therapies. This flow-cytometry-based procedure could also be rapidly translated into a clinically compatible assay.