Weight trends.

<p>(<b>A</b> and <b>B</b>) Mice inoculated with <i>M</i>. <i>abscessus</i> trended towards greater weight loss than mice inoculated with sterile thrombin/ fibrinogen. (N = 19–40 mice in each <i>M</i>. <i>abscessus</i> group, N = 6–16 mice in each sterile inoculum group). Mice receiving smooth morphotype inoculation that had some colony conversion to rough morphotype on either BALF, lung, or spleen cultures had trend towards greater weight loss and slower weight gain. (N = 8–10 for WT mice, 4–6 for CF mice). Data displayed as mean of group, pooled from 2–3 replicate experiments for each morphotype.</p

BALF macrophages.

<p>(<b>A</b>, <b>B</b>, <b>C</b> and <b>D</b>) Levels of BALF macrophages did not differ between groups. (N = 9–20 mice in each <i>M</i>. <i>abscessus</i> group, N = 3–8 mice in each sterile group). Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype.</p

Fibrin plug model.

<p>Thrombin and fibrinogen solutions, combined here on the bench top, form gelatinous fibrin plugs that retain the bacteria in the distal airways.</p

BALF neutrophil percentage.

<p>(<b>A</b>, <b>B</b>, <b>C</b> and <b>D</b>) Inoculation with rough morphotype causes greater BALF neutrophil percentage than smooth morphotype at 14 days post-inoculation in CF mice (<b>D</b>). Red circles indicate smooth morphotype inoculation with conversion to rough morphotype. Minimal BALF neutrophilia was seen after sterile thrombin/fibrinogen inoculation. (N = 9–20 mice in each <i>M</i>. <i>abscessus</i> group, N = 3–8 mice in each sterile group). * p< 0.05. Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype.</p

Pulmonary and splenic CFU.

<p>(<b>A</b>, <b>B</b>, <b>C</b> and <b>D</b>) Intratracheal inoculation with <i>M</i>. <i>abscessus</i> (smooth and rough morphotypes) suspended in fibrin plugs results in predominance of pulmonary infection with minimal systemic infection. (N = 9–20 mice in each group at 3 and 14 day time points). * p< 0.05. Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype.</p

BALF neutrophils.

<p>(<b>A</b>, <b>B</b>, <b>C</b> and <b>D</b>) Inoculation with rough morphotype causes greater BALF neutrophilia than smooth morphotype at 14 days post-inoculation in both WT (<b>B</b>) and CF (<b>D</b>) mice. Red circles indicate smooth morphotype inoculation with conversion to rough morphotype. Minimal BALF neutrophilia was seen after sterile thrombin/fibrinogen inoculation. (N = 4–20 mice in each <i>M</i>. <i>abscessus</i> group, N = 3–8 mice in each sterile group). * p< 0.05. Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype.</p

BALF cytokines.

<p>(<b>A</b> and <b>B</b>) BALF cytokines 3 days post-inoculation did not differ between groups. (N = 3–18 in each WT group, N = 5–6 in each CF group). Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype. Dashed line indicates lower limit of detection of kit.</p

Changes in pulmonary and splenic CFU over time.

<p>(<b>A</b> and <b>B</b>) Pulmonary infection wanes over time with both smooth and rough morphotypes and in both WT and CF mice. Data displayed as mean ± SEM of group, pooled from 2–3 replicate experiments for each morphotype.</p