Survival of TRAMP mice with (<i>Ncf1 */*</i>, n=23) and without (<i>Ncf1 +/+</i>, n=42) <i>Ncf1</i>^<i>m1J</i> mutation displayed as Kaplan-Meier curve, log rank test.

<p>Survival of TRAMP mice with (<i>Ncf1 */*</i>, n=23) and without (<i>Ncf1 +/+</i>, n=42) <i>Ncf1</i>^<i>m1J</i> mutation displayed as Kaplan-Meier curve, log rank test.</p

The Development of the B16-FLT3L tumors is restricted in the absence of functional NOX2 complex.

<p><b>A</b>) The weights of the B16-FLT3L tumors in <i>Ncf1</i>^<i>m1J</i> mutated (<i>Ncf1 */*</i>, n=7) and wild type (<i>Ncf1 +/+</i>, n=5) mice. All data presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084148#pone-0084148-g001" target="_blank">Figure 1</a> was collected d14 after tumor implantation. <b>B</b>) The total number of spleen cells in the <i>Ncf1</i>^<i>m1J</i> mutated (Ncf1*/*) and wild type (Ncf1 +/+) B16-FLT3L tumor bearing (FLT3L expressing tumor) and in naïve wild type mice (<i>Ncf1 +/+</i>, naïve). <b>C</b>) The percentages of B cells (B220+), dendritic cells (DC, CD11c-HIGH) neutrophils (Nf, Gr-1+) and T cells (CD3+) were gated from all splenocytes. Oxidative burst in splenic <b>D</b>) DC (CD11c-HIGH) and <b>E</b>) Nf (Gr-1+). The studied cell populations were phenotypically gated from all splenocytes. n=5 in all groups, bars represent mean values ±SEM, Mann-Whitney U test, p<0.05 is denoted with * and p<0.01 with **.</p

Lung parameters in response to BCG infection.

<p>(A) Lung/weight ratio of wild-type (n = 9), <i>Ncf1</i> mutant (n = 5) and <i>Ncf1</i> rescue (n = 8) mice without BCG infection and at 3 days and 4 weeks post infection. (B) Determination of alveolar space score (occupied lung tissue <i>vs.</i> free space) in lung sections at 4 weeks post infection. Data are represented as the mean of alveolar space score ± SD in 4 mice per group with at least 3 lobes analyzed per mouse. (C) Number of viable bacteria was determined at 3 days and 4 weeks following BCG infection. Data are shown as mean log of CFU per organ (±SEM; 3–5 mice per group). (D) iNOS protein expression in lung was detected by western blot 4 weeks after BCG infection. Results are expressed as mean ± SEM of relative units of iNOS/actin (n = 4, per group) after quantification by Image Quant software. (E) Nitrotyrosine quantification by ELISA was done in lungs, 4 weeks after BCG infection. Results are expressed as mean ± SEM of nM per lung (n = 4–5, per group). (***: p<0.001, **: p<0.05, *: p<0.01).</p

Cytokine and chemokine responses to BCG infection.

<p>TNF-α (A), IL-17 (B), IL-12p40 (C), IFN-γ (D), CXCL1 (E) and CCL5 (F) were assessed in lung homogenates obtained 3 days and 4 weeks after BCG infection. Results are presented as the mean ± SEM (n = 4–7 mice per group). (**: p<0.05, *: p<0.01).</p

Analysis of published cases of mycobacterial infections in CGD patients.

<p>Our literature research identified a total of 297 published cases of mycobacterial disease in CGD patients. (A) Mycobacterial species recovered in mycobacterial disease in CGD patients. (B) Clinical presentations of <i>Mycobacterium bovis</i> BCG and <i>Mycobacterium tuberculosis</i> infections in CGD patients. The numbers indicated on top of each column represent the percentage with respect to the total number of BCG or <i>M. tuberculosis</i> cases, respectively. The terms “systemic” refers to disseminated or to lung infections.</p

Impact of CGD mutation on mortality and weight loss in response to BCG infection.

<p><i>Ncf1</i> mutant (loss of function mutation in p47^phox), <i>Ncf1</i> rescue (expression of wild-type p47^phox in mononuclear phagocytes) with C57Bl/10.Q background, <i>Ncf1</i> mutant (loss of function mutation in p47^phox) with C57Bl/6 background, <i>Cybb</i><b>-</b>deficient and their respective wild-type controls were injected intravenously with BCG (10⁷ CFU). Survival was monitored over the 4 weeks period following BCG inoculation in (A) C57Bl/10.Q wild-type (n = 15), <i>Ncf1</i> mutant (n = 15) and <i>Ncf1</i> rescue (n = 11), in (C) C57Bl/6 wild-type (n = 7), <i>Ncf1</i> mutant (n = 6) and (E) C57Bl/6 wild-type (n = 7), <i>Cybb</i><b>-</b>deficient (n = 9) mice. (B, D and F) Body weight changes as a function of time after BCG inoculation. Survival (percent of initial number of mice) is shown in brackets. Statistics shown in the figures are the comparison between respective wild-type and <i>Ncf1</i> mutant or <i>Cybb</i><b>-</b>deficient (***: p<0.001, **: p<0.05, *: p<0.01) and the comparison between <i>Ncf1</i> mutant and rescue (§§§: p<0.001 and §§: p<0.01). Note that no significant differences were observed between wild-type and <i>Ncf1</i> rescue mice.</p