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Cells 2018, 7(6), 50; https://doi.org/10.3390/cells7060050

B Cells and B Cell Blasts Withstand Cryopreservation While Retaining Their Functionality for Producing Antibody

1
Research & Development Department, Cellular Technology Limited, Shaker Heights, OH 44122, USA
2
Institute of Anatomy and Cell Biology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, Germany
*
Author to whom correspondence should be addressed.
Received: 6 April 2018 / Revised: 2 May 2018 / Accepted: 22 May 2018 / Published: 31 May 2018
(This article belongs to the Special Issue Recent Advances in ELISPOT Research)
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Abstract

In individuals who have once developed humoral immunity to an infectious/foreign antigen, the antibodies present in their body can mediate instant protection when the antigen re-enters. Such antigen-specific antibodies can be readily detected in the serum. Long term humoral immunity is, however, also critically dependent on the ability of memory B cells to engage in a secondary antibody response upon re-exposure to the antigen. Antibody molecules in the body are short lived, having a half-life of weeks, while memory B cells have a life span of decades. Therefore, the presence of serum antibodies is not always a reliable indicator of B cell memory and comprehensive monitoring of humoral immunity requires that both serum antibodies and memory B cells be assessed. The prevailing view is that resting memory B cells and B cell blasts in peripheral blood mononuclear cells (PBMC) cannot be cryopreserved without losing their antibody secreting function, and regulated high throughput immune monitoring of B cell immunity is therefore confined to—and largely limited by—the need to test freshly isolated PBMC. Using optimized protocols for freezing and thawing of PBMC, and four color ImmunoSpot® analysis for the simultaneous detection of all immunoglobulin classes/subclasses we show here that both resting memory B cells and B cell blasts retain their ability to secrete antibody after thawing, and thus demonstrate the feasibility of B cell immune monitoring using cryopreserved PBMC. View Full-Text
Keywords: four color B cell ELISPOT; immune monitoring; freeze-thawing PBMC; plasma cells; antibody secretion; immunoglobulins; antibodies; immunoglobulin classes and subclasses; antibody-secreting cells; IgA; IgE; IgD; IgM; IgG1; IgG2; IgG3; IgG4; multiplex immune assay four color B cell ELISPOT; immune monitoring; freeze-thawing PBMC; plasma cells; antibody secretion; immunoglobulins; antibodies; immunoglobulin classes and subclasses; antibody-secreting cells; IgA; IgE; IgD; IgM; IgG1; IgG2; IgG3; IgG4; multiplex immune assay
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Fecher, P.; Caspell, R.; Naeem, V.; Karulin, A.Y.; Kuerten, S.; Lehmann, P.V. B Cells and B Cell Blasts Withstand Cryopreservation While Retaining Their Functionality for Producing Antibody. Cells 2018, 7, 50.

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