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. 2023 Mar 15;13:1125901. doi: 10.3389/fcimb.2023.1125901

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Copyright © 2023 Rowe, Lebman and Ohman

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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Effects of alginate production on the phagocytosis of P. aeruginosa. (A) Percentage of THP-1 macrophages that have phagocytized FRD1 and FRD1131 at 30 min. THP-1 macrophages were infected with GFP-expressing Alg⁺ FRD1 or Alg^- FRD1131 for 30 min. Percent phagocytosis indicates the percent of macrophages that are associated with GFP-expressing P. aeruginosa as determined by flow cytometry. Phagocytosis of FRD1 was significantly inhibited compared to FRD1131. Data are shown as mean and standard deviation of 3 experiments. (B) Number of bacteria phagocytized by THP-1 macrophages in 1 h. THP-1 macrophages were infected with FRD1, FRD1131 or FRD1131C’ (Alg⁺) for 1 h. CFU indicates the number of colony-forming units recovered from macrophages. Phagocytosis of FRD1 and FRD1131C’ was significantly inhibited compared to FRD1131. Data are shown as mean and standard deviation of a representative triplicate experiment. (C) Percentage of MH-S macrophages that have phagocytized bacteria at 30 min. MH-S macrophages were infected with GFP-expressing FRD1 or FRD1131 for 30 min. Percent phagocytosis indicates the percent of macrophages that are associated with GFP-expressing bacteria as detected by flow cytometry. Phagocytosis of FRD1 was significantly inhibited compared to FRD1131. Data are shown as mean and standard deviation of 3 separate experiments. (D) Number of bacteria phagocytized by MH-S macrophages after 1 h. MH-S macrophages were infected with FRD1, FRD1131 or FRD1131C’ for 1 h. CFU indicates the number of colony-forming units recovered from macrophages. Phagocytosis of Alg⁺ FRD1 and FRD1131C’ were significantly inhibited compared to Alg^- FRD1131. Data are shown as mean and standard deviation of 3 separate experiments. (E) Percentage of MH-S macrophages that have phagocytized opsonized bacteria after 30 min. MH-S macrophages were infected with GFP-expressing FRD or FRD1131 for 30 min in the presence of 0.5% human serum. Percent phagocytosis indicates the percent of macrophages that are associated with GFP-expressing bacteria as detected by flow cytometry. Opsonic phagocytosis of FRD1 was significantly inhibited compared to FRD1131. Data are shown as mean and standard deviation of 3 separate experiments. (F) Number of bacteria phagocytized by THP-1 macrophages after 1 hour in the presence of 0.5% human serum. THP-1 macrophages were infected with opsonized FRD1, FRD1131 or FRD1131C’ for 1 h in the presence of 0.5% human serum. CFU indicates the number of colony-forming units recovered from macrophages. Opsonic phagocytosis of Alg⁺ FRD1 and FRD1131C’ were significantly inhibited compared to Alg^- FRD1131. Data are shown as mean and standard deviation of a representative triplicate experiment. Statistical significance was determined by ANOVA (* P < 0.05, ** P < 0.01, *** P < 0.001).