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. 2019 Nov 27;9:17732. doi: 10.1038/s41598-019-54053-w

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© The Author(s) 2019

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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1A8 mAb potently suppressed BAL neutrophil numbers during SP infection. (A–C) Mice were inoculated with a low inoculum of SP (10⁵ CFU) and treated with 1A8 mAb or isotype control (ISO) one day prior and one day after SP infection. At day 2 post SP infection, bronchoalveolar lavage (BAL) was performed. (A) The total BAL cell count was increased in SP-infected mice and 1A8 treatment significantly reduced this response to control/uninfected levels. (B) Differential cell counts were performed and showed that BAL macrophages were not significantly altered by SP infection or 1A8 treatment. (C) BAL neutrophil numbers were markedly increased in response to SP infection and 1A8 mAb treatment significantly suppressed this response by over 90%. (D–F) Mice were inoculated with a high inoculum of SP (3 × 10⁶ CFU) and treated with 1A8 mAb or isotype control (ISO) one day prior and one day after SP infection. At day 1 post SP infection BAL was performed, and differential counts showed that (D) BAL macrophages were not significantly altered by SP infection or 1A8 treatment. (E) BAL neutrophil numbers were markedly increased in response to SP infection and 1A8 mAb treatment significantly suppressed this response by over 90%. (F) Assessment of net dsDNA levels in the BALF was used a marker for netosis, where levels were markedly increased by SP-ISO infection, and 1A8 mAb treatment significantly reduced dsDNA levels by over 90%. (G) Lung myeloperoxidase (MPO) activity was quantified as a marker for tissue neutrophil content, where increased activity in SP-ISO treated mice was significantly reduced by 1A8 mAb to control/uninfected levels. n = 5–6 ^*p < 0.05, one-way ANOVA.