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Optimisation of ex vivo memory B cell expansion/differentiation for interrogation of rare peripheral memory B cell subset responses

Muir, Luke, McKay, Paul F., Petrova, Velislava N., Klymenko, Oleksiy V., Kratochvil, Sven, Pinder, Christopher L., Kellam, Paul and Shattock, Robin J. (2018) Optimisation of ex vivo memory B cell expansion/differentiation for interrogation of rare peripheral memory B cell subset responses Wellcome Open Research, 2, 97. pp. 1-25.

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Background: Human memory B cells play a vital role in the long-term protection of the host from pathogenic re-challenge. In recent years the importance of a number of different memory B cell subsets that can be formed in response to vaccination or infection has started to become clear. To study memory B cell responses, cells can be cultured ex vivo, allowing for an increase in cell number and activation of these quiescent cells, providing sufficient quantities of each memory subset to enable full investigation of functionality. However, despite numerous papers being published demonstrating bulk memory B cell culture, we could find no literature on optimised conditions for the study of memory B cell subsets, such as IgM+ memory B cells.

Methods: Following a literature review, we carried out a large screen of memory B cell expansion conditions to identify the combination that induced the highest levels of memory B cell expansion. We subsequently used a novel Design of Experiments approach to finely tune the optimal memory B cell expansion and differentiation conditions for human memory B cell subsets. Finally, we characterised the resultant memory B cell subpopulations by IgH sequencing and flow cytometry.

Results: The application of specific optimised conditions induce multiple rounds of memory B cell proliferation equally across Ig isotypes, differentiation of memory B cells to antibody secreting cells, and importantly do not alter the Ig genotype of the stimulated cells. Conclusions: Overall, our data identify a memory B cell culture system that offers a robust platform for investigating the functionality of rare memory B cell subsets to infection and/or vaccination.

Item Type: Article
Divisions : Faculty of Engineering and Physical Sciences > Chemical and Process Engineering
Authors :
Muir, Luke
McKay, Paul F.
Petrova, Velislava N.
Klymenko, Oleksiy
Kratochvil, Sven
Pinder, Christopher L.
Kellam, Paul
Shattock, Robin J.
Date : 24 January 2018
Funders : Wellcome Trust, Engineering and Physical Sciences Research Council (EPSRC), Dormeur Investment Service Ltd
DOI : 10.12688/wellcomeopenres.11386.2
Copyright Disclaimer : Copyright: © 2018 Muir L et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Uncontrolled Keywords : Memory B cell subsets; Human; Vaccination; Ex vivo expansion/differentiation; Design of Experiments
Depositing User : Clive Harris
Date Deposited : 14 Oct 2019 08:31
Last Modified : 14 Oct 2019 08:31

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