Abstract
Based on the standard mouse phenotyping and intracellular cytokine panels, our work recently focused on establishing CyTOF workflows for the analysis of murine B cells, plasma cells, and regulatory T cells (Tregs). We will give a short update on challenges we experienced and solutions we came up with.
In order to achieve a deep phenotypic profiling of B cells and plasma cells, we used CyTOF analysis to investigate the expression of immunoglobulins, activation markers as well as co-stimulatory molecules in cells of gut-associated lymphoid tissues in our mouse models of inflammatory bowel disease. With the aim to obtain a detailed analysis of Tregs in the tumor microenvironment, we employed a subcutaneous tumor model in the Foxp3eGFP-Cre-ERT2 mouse strain. Tregs were identified with the Treg lineage marker Foxp3, CD25 and the presence of EGFP reporter expression, and suppressive markers expressed on Tregs such as CTLA-4, PD-1, ICOS and Perforin as well as cytokine production were determined.
Biosketch Selina Keppler
Selina currently is a Junior group leader at the Translational Cancer Center (TranslaTUM) at the Klinikum rechts der Isar in Munich. The Keppler lab is especially interested in the crosstalk of B cells with specialized inflammatory niches during autoimmunity, such as the gut or the kidney. In order to understand the complexity of autoimmune processes we combine imaging approaches with high-parametric flow cytometry, mass cytometry (CyTOF) and in vitro culture systems to define drivers of inflammation during homeostatic and inflammatory conditions.In addition to leading her research group, Selina is responsible for the training of users of the Core Facility of Cell Analysis in theory of flow cytometry, handling of the BDCanto and BDFortessa flow cytometers as well as multi-parametric panel design.
Biosketch Marc Rosenbaum
Marc is a senior post-doctoral researcher in the laboratory of Prof. Ruland at the Translational Cancer Center (TranslaTUM) at the Klinikum rechts der Isar in Munich. He has a long-standing interest in immunology, which he developed during studying Molecular Medicine at the University of Freiburg, the University of Western Australia, and the Max Planck Institute of Immunobiology and Epigenetics in Freiburg. Currently, he investigates the role of the CARD11-BCL10-MALT1 signaling complex in regulatory T cells. Besides, he trains users of the Core Facility of Cell Analysis in Amnis Imaging Flow Cytometry. In early 2020, he started working with the CyTOF technology and is interested in a detailed analysis of Tregs in the tumor microenvironment.
Keywords
mouse CyTOF panels, B cells, regulatory T cells
Contact
selina.keppler@tum.de
marc.rosenbaum@tum.de