In Situ Multiplex Immunofluorescence Analysis of Plasma Cell Myeloma Tissue Sections. (21st September 2018)

• Record Type: Journal Article
• Authors: Sood, Anup; Ginty, Fiona; Chadwick, Chrystal; Janz, Siegfried; Holman, Carol
• Is Part Of: American journal of clinical pathology. Volume 150(2018)Supplement 1
• Page Start: S161
• Page End: S162
• Publication Date: 2018-09-21
• Subjects: Diagnosis, Laboratory -- Periodicals; Pathology -- Periodicals; 616.07
• DOI: 10.1093/ajcp/aqy112.375
• Languages: English
• ISSNs: 0002-9173

Abstract

Abstract: Plasma cell myeloma (PCM), with a highly heterogeneous clinical behavior and response to therapy, accounts for approximately 10% of hematologic malignancies. Current diagnostic and prognostic tests include serum-based assays, multiparametric flow cytometric analysis of bone marrow aspirates, morphologic and immunohistochemical analysis of bone marrow sections, cytogenetic analysis, and in vivo imaging. None of these tools allow for determination of expression of large numbers of antigens on a single myeloma cell in spatial context, which may be useful in order to better understand disease progression and therapy response. In a pilot study, we sought to evaluate the performance of a new in situ hyperplex immunofluorescence platform, Cell-DIVE, that allows spatial and subcellular localization of over 60 markers on three bone marrow samples (two non-decalcified clot sections and one decalcified core biopsy) from PCM cases. The platform utilizes a signal cycling process wherein repeated cycles of staining with fluorophore-conjugated antibodies, imaging, and signal removal are performed on the same tissue sections. Postcycling, images from multiple cycles are registered, corrected for tissue autofluorescence, and segmented into single cells and subcellular compartments using compartment-specific markers. To demonstrate feasibility, a panel of five markers that included plasma cell markers (CD138, CD45), markers of clonality (kappa and lambda light chains), and the proliferation marker Ki-67 were evaluated on a single histologic section. A virtual H&E image was also generated from the same tissue section. Both clot and core biopsy samples showed staining patterns similar to immunohistochemistry—the PCM cells expressed CD138 and either kappa or lambda light chains but lacked CD45. The cellular localization of staining was also as expected, with CD138 displaying surface staining, light chains displaying cytoplasmic staining, and Ki-67 displaying nuclear staining. This new immunofluorescence platform blends the strengths of immunohistochemistry (preservation of tissue architecture) with those of flow cytometry (analysis of many antigens on the same cell). Our preliminary data suggest that the in situ hyperplex immunofluorescence platform is a powerful new approach to study PCM biology, which allows for preservation of the spatial context of malignant plasma cells relative to a heterogeneous tumor microenvironment, assessing more antigens on a single cell than has been possible previously. Additionally, this platform gives us a new method for assessing heterogeneity at the level of the tumor cell clone—an important research tool that shows great promise for synergizing with other newly emerging, single cell-based technologies.