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. 2020 Aug 6;10:13274. doi: 10.1038/s41598-020-70203-x

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© The Author(s) 2020

Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Generation of pathogenic anti-AQP4 recombinant antibodies derived from patients’ plasmablasts. (A) CD3^- CD19^int CD138⁺ plasmablasts are isolated from patients’ CSF lymphocytes. (B) The highest percentage of plasmablasts is observed in the sample derived from patient 4 (arrow head). (C) The interclonal diversity of V regions of eight clones which showed positive binding to AQP4 are shown in red. (D) Immunoreactivity to AQP4-expressing HEK293 cells is assessed by FACS analysis. The clones GK15, 41, 74 and 89 show remarkable binding to AQP4. (E) The immunoreactivity of clones GK89 to AQP4-expressing HEK293 cells is shown. (F) The percentage of damaged cells is measured by LDH assay. Clone GK89 has the highest capacity to induce complement-dependent cytotoxicity (arrow head). Error bars indicate SEM.