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. 2001 Aug;69(8):4823–4830. doi: 10.1128/IAI.69.8.4823-4830.2001

FIG. 5.

FIG. 5

Anti-Omp25 antibodies specifically reverse the inhibition of TNF-α in human macrophage infection. (A and B) WT B. suis (A) and Δomp25 B. suis (B) were incubated for 30 min with anti-Omp25 or anti-Omp31 (dilution, 1/5) or with an irrelevant IgG2a. The bacteria were then extensively washed and incubated for 30 min with an anti-mouse FITC-F(ab′)2 fragment. Their fluorescence was then measured by flow cytometry. 1, WT B. suis plus anti-CD14 (irrelevant IgG2a) plus anti-mouse FITC-F(ab′)2; 2, WT B. suis plus anti-Omp25 plus anti-mouse FITC-F(ab′)2; 3, WT B. suis plus anti-Omp31 plus anti-mouse FITC-F(ab′)2. FL1-H, fluorescence arbitrary units. (C) WT B. suis was incubated for 30 min with different dilutions of anti-Omp25 (●) or anti-Omp31 (○) antibodies or with medium alone. (x axis, percentages of hybridoma supernatants [HS] during incubation.) The bacteria were then washed and used to infect THP-1 cells as described in Materials and Methods (MOI, 20). Seven hours later, macrophage culture supernatants were harvested and TNF-α concentrations in cell supernatants were measured (y axis). The values shown are the means ± SD of four different experiments. In this set of experiment, noninfected cells released 73 ± 10 pg of TNF-α per ml. Differences in TNF-α production between cells infected with WT B. suis, anti-Omp25-treated WT B. suis, and anti Omp31-treated B. suis were analyzed by paired t tests.