Sensitive intranuclear flow cytometric quantification of IRF4 protein in multiple myeloma and normal human
Ashni A Vora1, Phoebe K Mondala1, Caitlin Costello2,3
1Division of Regenerative Medicine, Department of Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
Insights
Quantifying Interferon regulatory factor 4 (IRF4) protein in immune cells using flow cytometry can help predict multiple myeloma severity and treatment response. This method offers a potential prognostic tool for personalized cancer therapy.
Area of Science:
- Immunology
- Molecular Biology
- Oncology
Background:
- Interferon regulatory factor 4 (IRF4) is a key transcription factor in immune cell development.
- IRF4 plays a critical role in the pathogenesis of multiple myeloma, a hematologic malignancy.
Purpose of the Study:
- To describe a flow cytometry-based protocol for measuring IRF4 protein expression.
- To evaluate the potential of IRF4 quantification as a prognostic biomarker in multiple myeloma.
Main Methods:
- Combined cell surface and intranuclear staining using fluorescent antibodies.
- Flow cytometry for protein quantification in myeloma and normal immune cells.
Main Results:
- The protocol enables measurement of IRF4 protein levels within specific immune cell populations.
- IRF4 quantification shows potential as a prognostic indicator for disease severity and therapy response.
Conclusions:
- IRF4 protein measurement via flow cytometry is a feasible approach.
- This assay could serve as a valuable prognostic tool and be adapted for clinical use in multiple myeloma management.
Abstract:
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).
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