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Multiplex Immunofluorescence of Bone Marrow Core Biopsies: Visualizing the Bone Marrow Immune Contexture
Denise K Walters1, Diane F Jelinek1
1Department of Immunology, College of Medicine and Science, Mayo Clinic, Rochester, Minnesota and Scottsdale, Arizona.
Insights
Multiplex immunofluorescence analysis is now achievable in bone marrow core biopsies. This breakthrough allows researchers to visualize immune cells near tumor cells, aiding understanding of cancer progression in monoclonal gammopathies.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Visualizing immune and tumor cell proximity offers insights into cancer biology.
- Multiplex immunofluorescence (MIF) is a powerful technique for analyzing multiple targets in tissue sections.
- MIF has not been previously applied to bone marrow core biopsies.
Purpose of the Study:
- To adapt multiplex immunofluorescence analysis for bone marrow core biopsies.
- To investigate the spatial relationship between immune cells and malignant plasma cells in multiple myeloma.
- To enable novel analyses of the immune microenvironment in monoclonal gammopathies.
Main Methods:
- Developed a strategy to quench autofluorescence in bone marrow tissues.
- Implemented a multiplex staining protocol using a stripping solution for sequential antibody removal.
- Validated the efficiency and preservation of antigenicity and tissue integrity.
Main Results:
- Successfully adapted multiplex immunofluorescence for bone marrow core biopsies.
- The developed method efficiently visualizes multiple epitopes without compromising tissue quality.
- Demonstrated the feasibility of analyzing immune cell proximity to malignant plasma cells.
Conclusions:
- Multiplex immunofluorescence is now a viable technique for bone marrow core biopsies.
- This advancement opens new avenues for studying immune cell interactions in cancer.
- Facilitates deeper understanding of immune contexture in monoclonal gammopathies and disease progression.
Abstract:
The ability to visualize and quantify the spatial arrangement and geographic proximity of immune cells with tumor cells provides valuable insight into the complex mechanisms underlying cancer biology and progression. Multiplexing, which involves immunofluorescence labeling and the visualization of multiple epitopes within formalin-fixed paraffin embedded tissue sections, is a methodology that is being increasingly employed. Despite the power of immunofluorescence multiplex analysis, application of this technology to bone marrow core biopsies has not yet been realized. Given our specific long term goal to identify immune cells in proximity to bone marrow malignant plasma cells in multiple myeloma patients, we describe in this study adaptation of multiplex immunofluorescence analysis to this tissue. We first identified a blocking strategy that quenched autofluorescence. We next employed a multiplex strategy that uses a simple stripping solution to remove primary and secondary antibodies prior to subsequent rounds of staining. This method was found to be highly efficient and did not significantly alter antigenicity or tissue integrity. Our studies illustrate for the first time that immunofluorescence multiplexing is achievable in bone marrow core biopsies and will provide a novel opportunity to analyze the role of the immune contexture in disease progression of the monoclonal gammopathies.
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