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Master Thesis Project – Mass cytometry –

Published: 21. Aug 2018

Aim: Development of a mass cytometry assay for deep phenotyping of plasma cells

Background: Plasma cells are highly differentiated, antibody-secreting B lymphocytes. They provide specific humoral immunity and are thereby intimately linked to vaccine development, research in autoimmunity and immune memory. Therapeutic targeting of plasma cells is considered for the treatment of antibody-mediated diseases.

We study plasma cells, and their immediate precursors, the plasmablasts, in the blood and bone marrow as correlates of acute and established immune responses, respectively. Plasmablasts and plasma cells detectable in blood and bone marrow show considerable heterogeneity, e.g. with respect to markers of differentiation state, cell migration and adhesion, proliferation, apoptosis and type of secreted antibody.

The Project: You will establish a mass cytometry (CyTOF) assay for deep phenotyping of plasma cells. By these means you will generate maps of plasmablasts and plasma cells from healthy individuals, creating a basis for analyzing specific changes in autoimmune diseases such as rheumatoid arthritis and Systemic Lupus Erythematosus, and for studying plasma cell diversification in vitro. Results are expected to provide new insight into plasma cell differentiation in humans.

In the lab: You will learn about plasma cells and how to design a complex mass cytometry panel and workflow, generate  and analyze the data using advanced computational tools suited to evaluate high-dimensional cytometry data. The project will further involve heavy metal conjugation of antibodies and isolation of plasma cells from human blood and BM involving magnetic and flow cytometric cell sorting (MACS/FACS).

The DRFZ is an equal opportunity/affirmative action employer, committed to excellence through diversity, and strongly encourages applications and nominations of persons of color, women, and members of under-represented groups.
Individuals with disabilities will be given priority, if qualified accordingly.

Contact
Dr. rer. nat. Henrik Mei
Mass cytometry
Deutsches Rheuma-Forschungszentrum Berlin (DRFZ) Charitéplatz 1, 10117 Berlin, Germany
Phone: +49 030 28 460 774 E-mail: mei@drfz.de


References & Reading

Mass cytometry

  • C. Bendall, G. P. Nolan, M. Roederer, P. K. Chattopadhyay, A deep profiler’s guide to cytometry. Trends Immunol, (2012).
  • D. Tanner, V. I. Baranov, O. I. Ornatsky, D. R. Bandura, T. C. George, An introduction to mass cytometry: fundamentals and applications. Cancer immunology, immunotherapy : CII 62, 955- 965 (2013).
  • H. E. Mei, M. D. Leipold, H. T. Maecker, Platinum-conjugated antibodies for application in mass cytom Cytometry A, (2015).
  • H. E. Mei, M. D. Leipold, A. R. Schulz, C. Chester, H. T. Maecker, Barcoding of live human peripheral blood mononuclear cells for multiplexed mass cytom J Immunol 194, 2022-2031 (2015).

Plasma cells

  • T. Yoshida et , Memory B and memory plasma cells. Immunol Rev 237, 117-139 (2010).
  • H. E. Mei et , A unique population of IgG-expressing plasma cells lacking CD19 is enriched in human bone marrow. Blood 125, 1739-1748 (2015).
  • H. E. Mei et , Steady-state generation of mucosal IgA+ plasmablasts is not abrogated by B-cell depletion therapy with rituximab. Blood 116, 5181-5190 (2010).
  • H. E. Mei et , Blood-borne human plasma cells in steady state are derived from mucosal immune responses. Blood 113, 2461-2469 (2009).
Zentrallabor Massenzytometrie Dr.rer. nat. Henrik Mei Tel +49 (0)30 28460 - mei@drfz.de