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. 2021 May 11;12:573266. doi: 10.3389/fimmu.2021.573266

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Copyright © 2021 Cole, Angyal, Emes, Mitchell, Dickman and Dockrell

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Pneumolysin leads to repression TNF and PTGS2 production. Monocyte derived macrophages (MDMs) were challenged with either S. pneumoniae, the isogenic pneumolysin negative mutant (Δply), or mock-infected with phosphate buffered saline (MI). After 3 h incubation RNA was extracted from MDMs and RT-qPCR performed to measure the abundance of (A) TNF, (B) PTGS2, (C) C-JUN, (D) IL1RN and (G) NR4A2 mRNA. The Bar chart represents the ΔΔCT fold change for each bacterial challenge demonstrating significantly higher abundance of TNF, PTGS2, C-JUN and IL1RN mRNA following challenge with Δply. (G) There was no significant difference in the abundance of NR4A2 mRNA in response to either bacterial challenge. (E) A representative western blot after probing with anti-PTGS2 antibody is shown, with actin used as a loading control. The blot is representative of four independent experiments. (F) Densitometry was performed to estimate the PTGS2/actin and normalised to mock infected sample. The bar charts represent mean and Standard Deviation, n= 10 for panel (A, B). n=6 for panel (C) and n=7 for panel (D, G); *p < 0.05, **p < 0.01.