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The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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CD3-positive B cells: a storage-dependent phenomenon.

Angela Nagel1, Christian Möbs2, Hartmann Raifer3

  • 1Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany; Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.

Plos One
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PubMed
Summary
This summary is machine-generated.

Human B cells can show T cell antigen CD3 after ex vivo blood storage. This antigen exchange depends on temperature and duration, potentially misclassifying cell types in studies.

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Area of Science:

  • Immunology
  • Cell Biology
  • Clinical Research

Background:

  • Clinical studies often require extended handling of human specimens, including blood samples for shipping and analysis.
  • Ex vivo storage of blood samples can alter antigen expression on lymphocyte surfaces, impacting experimental outcomes.

Purpose of the Study:

  • To investigate the effect of ex vivo blood storage on antigen expression in human lymphocytes.
  • To determine the mechanism and influencing factors of observed antigen changes on B cells.

Main Methods:

  • Analysis of human peripheral blood samples subjected to varying ex vivo storage conditions (temperature, duration).
  • Flow cytometry and phenotypic analysis to detect T cell antigen CD3 expression on B cells.

Main Results:

  • The T cell antigen CD3 was detected on the surface of human B cells following ex vivo storage.
  • The extent of CD3 detection on B cells was significantly dependent on storage temperature and time post-collection.
  • This phenomenon is attributed to contact-dependent antigen exchange between T and B cells, not de novo production.

Conclusions:

  • Ex vivo storage can lead to the misclassification of peripheral blood cell subsets due to antigen transfer.
  • Minimizing ex vivo storage time for peripheral blood mononuclear cells (PBMCs) is crucial for accurate in vivo representation.
  • Standardizing and specifying storage conditions are essential for ensuring reproducibility in multicenter studies and research.