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. 2020 Apr 15;8:49. doi: 10.1186/s40478-020-00920-x

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Antibody mediated complement activation of serum samples from rats immunized with AQP4 mimotopes. a: Serum AQP4 antibodies from an AQP4 antibody positive NMOSD patient (a) and the mouse monoclonal E5415A AQP4 antibody were able to activate the complement cascade in the presence of active rat complement on rat-AQP4-EmGFP (green) expressing HEK293 cells resulting in TCC deposition (red) and cell death (DAPI, blue). a and b are positive controls (pos co 1 and 2). c-f: Sera from rats immunized with mimotope-AQP4_268–285/CFA (mimotopes were IV-04 inducing rat AQP4-spec antibodies in a titer of 1:160, III-01 inducing rat AQP4-spec antibodies in a titer of 1:160, I-04 inducing rat AQP4-spec antibodies in a titer of 1:320, and I-13 inducing rat AQP4-spec antibodies in a titer of 1:320) were not able to activate the complement cascade as shown by fewer DAPI (blue) positive cells and no TCC (red) deposition. g: Serum from a III-01-AQP4_268–285/CFA immunized rat with an antibody titer of zero was used as a first negative control (neg co 1) and shows no antibody mediated complement activation. h: Same serum as in A in the presence of heat-inactivated rat complement (second negative control (neg co 2) does not activate the complement cascade and shows no TCC (red) formation. Scale bar = 100 μm