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. 2023 Sep 8;42:235. doi: 10.1186/s13046-023-02798-8

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© The Author(s) 2023

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 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 — Ascites dialysis partially reverses inhibitory properties caused by non-protein and heat resistant components. (A) Graphical illustration of protein depletion in ascites. Ascites sample was processed via ultracentrifugal filtration and protein-rich retentate and protein-less permeate are collected. (B) Composition of ascites before and after protein depletion. Heatmap showing altered composition of protein-depleted ascites samples and protein-rich retentates. (C) Electrophoresis of ascites sample. Representative electrophoresis blot of ascites sample 16 and corresponding permeate. (D) Proliferation of activated T cells in presence of protein-depleted ascites. Prestained donor T-cells were stimulated using CD2/CD3/CD28 activator complex (1:40) in protein-less ascites permeate. (E) NK ADCC in presence of ascites permeate. Resting NK cells were coincubated in 1:1 ratio with IGROV1 cells with addition of ADCC-inducing anti-EGFR antibody Cetuximab in presence of unmodified or protein-less ascites permeate. (F) NK ADCC in presence of heat inactivated ascites permeate. Resting NK cells were coincubated in 1:1 ratio with IGROV1-cells and ADCC-inducing anti-EGFR antibody Cetuximab with protein-depleted ascites permeate which was heat inactivated at 90 °C for 1 h. (G) Graphical illustration of ascites dialysis in cell culture medium. Ascites samples were processed in medium overnight using 1 kDa or 25 kDa cutoff dialysis columns. (H) Composition of ascites before and after dialysis. Heatmap showing altered composition of ascites samples processed by dialysis. (I) NK ADCC in presence of dialyzed ascites. Resting NK cells were coincubated in 1:1 ratio with IGROV1 cells with addition of ADCC-inducing anti-EGFR antibody Cetuximab in presence of unmodified or dialyzed ascites. After 6 h expression of CD107a on NK cells was determined by flow cytometry. Each datapoint represents one healthy donor. For significance testing ordinary one-way ANOVA followed by Dunnet posthoc test (4.D) and two-way ANOVA followed by Sidak posthoc test was used (4.E, F and I). ns (non-significant), * (p < 0.05), ** (p < 0.01), **** (p < 0.0001)