Monitoring B cell alloresponses in rats.

Louisa Steines, Mona Scharf, Petra Hoffmann, Antonia Schuster, Bernhard Banas, Tobias Bergler

Journal of immunological methods 2022 Feb

filter terms:

Antibody-mediated rejection is a major cause of graft failure in organ transplantation. For this reason, B cell responses are of particular interest to transplantation research. Rats are important model organisms for transplant studies, but B cell alloimmune assays and B cell subset markers are poorly established in rats. We alloimmunized rats by donor blood injection using the high responder rat strain combination Brown Norway (donor) and Lewis (recipient) rats. Using splenocytes from alloimmunized and control rats, we established assays to assess allospecific B cell proliferation and the capacity to generate allospecific B memory cells and alloantibody-secreting cells after antigenic rechallenge in vitro using a mixed lymphocyte reaction. Furthermore, we defined a simple gating and sorting strategy for pre- and post-germinal center follicular B cells, as well as non-switched and switched plasmablasts. Our protocols for assessing B cell alloresponses and B cell subsets in rats may help to accelerate research into the role of B cells and manipulation of humoral alloresponses in transplant research. Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.

Citation

Louisa Steines, Mona Scharf, Petra Hoffmann, Antonia Schuster, Bernhard Banas, Tobias Bergler. Monitoring B cell alloresponses in rats. Journal of immunological methods. 2022 Feb;501:113212