Bri Munnich is a research technician for the Krysiak Lab at Washington University in Saint Louis. The focus of our lab is to develop a method to isolate and characterize Hodgkin and Reed-Sternberg cells using a size based microfilter to isolate the larger cells of interest from the periphery found in the lymph node. Previously, Bri worked in industry for an ingredient company, Futureceuticals, as a part of their research and development team creating optimal blends of phytonutrients to reduce gut inflammation associated with IBS and Crohn’s Disease. She also acted as an adjunct instructor at Olivet Nazarene University, her alma mater, working with students in the cellular biology and biochemistry labs prior to coming to WashU. She plans to pursue her doctorate in pathobiology and continue working in academic research.
Brianna Munnicha, Becca Browna, Abdulrahman Saadallab, Richard Cotea, Todd Fehnigera, Siddarth Rawala, Kilannin Krysiaka
aWashington University in St. Louis School of Medicine, Saint Louis, MO, United States; bARUP Laboratories, Salt Lake City, UT, United States
Hodgkin lymphoma (HL) is a rare B-cell derived hematologic malignancy that affects the lymph nodes. Large, multinucleated Reed-Sternberg (50-100 µm) and mononuclear Hodgkin cells (20-30 µm) are pathognomonic features of HL. These cells are rare and associated with T-cell aggregates making isolation and characterization challenging. Previous methods employed to isolate Hodgkin and Reed-Sternberg (HRS) cells are technically challenging, time consuming, and produce limited outputs unsuitable for clinical assays, making an alternative methodology imperative for the continued study of HL.
To address these challenges, we turned to a size-based microfiltration system developed for the capture of circulating tumor cells (CTC) from peripheral blood. The system is designed to allow red blood cells, platelets and leukocytes (< 8 µm) to pass through while capturing and fixing CTCs (~10-16 µm) on the microfilter for on-chip downstream analysis. This process is rapid (< 10 min) and has established efficacy in the capture of rare CTCs across various malignancies. Additionally, protocols for immunofluorescence, molecular analysis, and fluorescence in situ hybridization (FISH) have been established for cells captured on the microfilter. Due to the large size of HRS cells, we hypothesized that this system could be utilized to enrich HRS cells from HL affected lymph nodes for use in downstream assays. Using L428s, a HL cell line, immunofluorescent staining, and QuPath, a digital pathology software, we aim to illustrate the degree of enrichment achievable on the microfilter and optimize this process for use with clinical samples.