Additional file 1: of Association between six-minute walk test parameters and the health-related quality of life in patients with pulmonary Mycobacterium avium complex disease

Supplemental analyses. Table S1. Clinical characteristics of patients with pulmonary Mycobacterium avium complex disease in the never smoker group (n = 92). Table S2. Results of the six-minute walk test for patients with pulmonary Mycobacterium avium complex disease in the never smoker group (n = 92). Table S3. Spearman’s correlations among six-minute walk test parameters and clinical parameters for patients with pulmonary Mycobacterium avium complex disease in the never smoker group (n = 92). Table S4. Spearman’s correlations among six-minute walk test parameters and 36-Item Short Form Health Survey and St George’s Respiratory Questionnaire scores for patients with pulmonary Mycobacterium avium complex disease in the never smoker group (n = 92). Table S5. Multivariate analysis for predictors of 36-Item Short Form Health Survey and St George’s Respiratory Questionnaire scores for patients with pulmonary Mycobacterium avium complex disease in the never smoker group (n = 92). Table S6. Comparisons of six-minute walk test parameters and 36-Item Short Form Health Survey and St George’s Respiratory Questionnaire scores with or without cavitary lesions (n = 103). Table S7. Comparisons of clinical characteristics in pulmonary Mycobacterium avium complex disease patients between male and female (n = 103). Table S8. Multivariate analysis for predictors of 36-Item Short Form Health Survey and St George’s Respiratory Questionnaire scores for patients with pulmonary Mycobacterium avium complex disease in female alone (n = 80). Table S9. Comparisons of 36-Item Short Form Health Survey and St George’s Respiratory Questionnaire scores with or without several clinical parameters (n = 103). (DOCX 55 kb

Additional file 1: Table S1. of The efficacy, safety, and feasibility of inhaled amikacin for the treatment of difficult-to-treat non-tuberculous mycobacterial lung diseases

Five different conditions were tried for the nebulization of amikacin sulphate. (DOCX 24 kb

Exotic hadrons bend the rules

The equipment used to measure the aerosolized particle sizes (Mastersizer 2000: Malvern Instruments Ltd., Worcestershire, UK). (DOCX 251 kb

Additional file 2: Table S2. of The efficacy, safety, and feasibility of inhaled amikacin for the treatment of difficult-to-treat non-tuberculous mycobacterial lung diseases

Nebulizing time and speed, final remaining dose and the particle size in the compressor nebulizer. A digital weighing scale was used to measure the mean nebulizing speed and the final remaining dose. The particle sizes were measured using a Mastersizer 2000 (Malvern Instruments Ltd., Worcestershire, UK). (DOCX 186 kb

Clarithromycin expands CD11b⁺Gr-1⁺ cells via the STAT3/Bv8 axis to ameliorate lethal endotoxic shock and post-influenza bacterial pneumonia

<div><p>Macrolides are used to treat various inflammatory diseases owing to their immunomodulatory properties; however, little is known about their precise mechanism of action. In this study, we investigated the functional significance of the expansion of myeloid-derived suppressor cell (MDSC)-like CD11b⁺Gr-1⁺ cells in response to the macrolide antibiotic clarithromycin (CAM) in mouse models of shock and post-influenza pneumococcal pneumonia as well as in humans. Intraperitoneal administration of CAM markedly expanded splenic and lung CD11b⁺Gr-1⁺ cell populations in naïve mice. Notably, CAM pretreatment enhanced survival in a mouse model of lipopolysaccharide (LPS)-induced shock. In addition, adoptive transfer of CAM-treated CD11b⁺Gr-1⁺ cells protected mice against LPS-induced lethality via increased IL-10 expression. CAM also improved survival in post-influenza, CAM-resistant pneumococcal pneumonia, with improved lung pathology as well as decreased interferon (IFN)-γ and increased IL-10 levels. Adoptive transfer of CAM-treated CD11b⁺Gr-1⁺ cells protected mice from post-influenza pneumococcal pneumonia. Further analysis revealed that the CAM-induced CD11b⁺Gr-1⁺ cell expansion was dependent on STAT3-mediated Bv8 production and may be facilitated by the presence of gut commensal microbiota. Lastly, an analysis of peripheral blood obtained from healthy volunteers following oral CAM administration showed a trend toward the expansion of human MDSC-like cells (Lineage⁻HLA-DR⁻CD11b⁺CD33⁺) with increased arginase 1 mRNA expression. Thus, CAM promoted the expansion of a unique population of immunosuppressive CD11b⁺Gr-1⁺ cells essential for the immunomodulatory properties of macrolides.</p></div

CAM-treated CD11b⁺Gr-1⁺ cells exhibit an immunosuppressive phenotype.

<p>(A) Top 25 upregulated and downregulated genes determined by a microarray analysis in splenic CD11b⁺Gr-1⁺ cells sorted from vehicle- and CAM-treated mice. *<i>Stxbp6</i> (chromosome 12:45956210–46175345). **<i>Stxbp6</i> (chromosome 12:45953470–45956090). Results are presented as fold changes relative to the expression levels of each gene in vehicle-treated CD11b⁺Gr-1⁺ cells. (B) Arginase activity in the spleen of vehicle- and CAM-treated mice (n = 4 per group). N.D., not detected. (C) Immunofluorescence staining of Gr-1 and arginase-1 in the lungs of mice treated with CAM daily for three consecutive days (n = 4 per group). Scale bar, 200 μm. (D) The concentration of nitric oxide (NO) in spleen extracts of vehicle- and CAM-treated mice (n = 4 per group) ***<i>p</i> < 0.001 by the Mann–Whitney U-test. (E) Expression of the surface marker CD244 on splenic CD11b⁺Ly-6G⁺ cells determined by flow cytometry (n = 4 per group). (F–H) Cytokine profile of the culture supernatant from bone marrow-derived macrophages (BMDMs) with or without equal numbers of vehicle-treated or CAM-treated CD11b⁺Gr-1⁺ cells (5 × 10⁵ cells) in the spleen: TNF-α (F), IFN-γ (G), and IL-10 (H). Representative data for three independent experiments are shown. Data are expressed as the mean ± SEM. ***<i>p</i> < 0.001 by a one-way ANOVA with Tukey’s multiple comparison tests.</p

Additional file 5: Figure S2. of The efficacy, safety, and feasibility of inhaled amikacin for the treatment of difficult-to-treat non-tuberculous mycobacterial lung diseases

Particle sizes and solution volumes in condition 1, condition 2 and condition 5. The aerosolized particle sizes were measured using a Mastersizer 2000 (Malvern Instruments Ltd., Worcestershire, UK). (DOCX 243 kb

Additional file 1: Table S1. of Clinical efficacy and safety of multidrug therapy including thrice weekly intravenous amikacin administration for Mycobacterium abscessus pulmonary disease in outpatient settings: a case series

Comparison of patient characteristics between patients receiving AMK and patients not receiving AMK (DOCX 14 kb

CAM ameliorates LPS-endotoxin shock via the essential contribution of CD11b⁺Gr-1⁺ cells.

<p>(A) Survival rate for LPS (50 mg/kg)-endotoxin shock in mice pretreated with vehicle or CAM (100 mg/day) daily for three consecutive days (n = 36 per group). *<i>p</i> = 0.0009 by the log-rank test. (B–D) Cytokine profiles in serum 12 h after LPS challenge in vehicle- or CAM-treated mice: TNF-α (B), IFN-γ (C), and IL-10 (D). (n = 5–6 per group). Data are represented as the mean ± SEM. **<i>p</i> < 0.01. ***<i>p</i> < 0.001 by the Mann–Whitney U-tests. (E and F) Representative two-parameter dot plots of CD11b⁺Gr-1⁺ cells in the spleen (E) and lungs (F) of mice intraperitoneally treated with vehicle or CAM (100 mg/day) daily for three consecutive days, followed by intraperitoneal injection with PBS or LPS (50 mg/kg) (n = 4 per group). (G) Quantification of CD11b⁺Gr-1⁺ cells in the spleen and lungs sorted from intraperitoneally vehicle- and CAM-treated (once a day for 3 days), followed by intraperitoneally LPS-treated mice (n = 4 per group). *<i>p</i> < 0.05, **<i>p</i> < 0.01 by Mann–Whitney U-tests. (H) Survival rate for LPS-endotoxin shock in vehicle- and CAM-injected mice pretreated with either anti-Gr-1 antibody (250 μg/mouse) or control IgG (n = 20–21 per group) 24 h before LPS challenge. *<i>p</i> = 0.0128 by the log-rank test. (I) Survival rate for LPS-endotoxin shock in vehicle- and CAM-injected mice pretreated with either anti-Gr-1 antibody (250 μg/mouse) or control IgG (n = 25–26 per group) 1 h before initiation of CAM treatment (i.e., 73 h before LPS challenge). Combined data for two independent experiments are shown. ***<i>p</i> < 0.001 by the log-rank test. (J) Adoptive transfer of CAM-treated CD11b⁺Gr-1⁺ cells improved the survival rate in LPS endotoxin shock (n = 24 per group). *<i>p</i> = 0.0023 by the log-rank test. (K-M) TNF-α (K), IFN-γ (L), and IL-10 (M) levels in serum at 12 h after intraperitoneal LPS injection (n = 5–6 per group). Data are presented as the mean ± SEM. *<i>p</i> < 0.05. **<i>p</i> < 0.01. ***<i>p</i> < 0.001 by the Mann–Whitney U-tests.</p

CAM improves survival in post-influenza pneumococcal pneumonia via an essential contribution of CD11b⁺Gr-1⁺ cells.

<p>(A) Survival rate of post-influenza pneumococcal pneumonia mice treated with vehicle, ampicillin (ABPC) (100 mg/kg), or clarithromycin (CAM) (100 mg/kg) (n = 37 per group). *<i>p</i> < 0.05, ***<i>p</i> < 0.001 by the log-rank test. (B) Cell counts in bronchoalveolar lavage fluid (BALF) obtained from mice with post-influenza pneumococcal pneumonia treated with vehicle, ABPC (100 mg/kg), or CAM (100 mg/kg) (n = 7–8 per group). Data are presented as the mean ± SEM. *<i>p</i> < 0.05. **<i>p</i> < 0.01, ***<i>p</i> < 0.001 by a two-way ANOVA with Tukey’s multiple comparison tests. (C and D) Bacterial load in the lungs (C) and blood (D) of post-influenza pneumococcal pneumonia mice treated with vehicle, ABPC (100 mg/kg), or CAM (100 mg/kg) at 18 and 36 h after pneumococcal infection (n = 15–16 per group). Data are presented as the mean ± SEM. *<i>p</i> < 0.05 by a two-way ANOVA with Tukey’s multiple comparison tests. (E) Lung H&E staining at 48 h after pneumococcal infection. Representative data for 5 mice per group are shown. Scale bar, 100 μm. (F–I) Levels of IFN-γ (F) and IL-10 (G) in BALF were measured by ELISA. The levels of IFN-γ (H) and IL-10 (I) in serum were measured by ELISA (n = 7–8 per group). Data are presented as the mean ± SEM. *<i>p</i> < 0.05. **<i>p</i> < 0.01. ***<i>p</i> < 0.001 by a two-way ANOVA with Tukey’s multiple comparison tests. (J) Survival rate of mice following adoptive transfer of CD11b⁺Gr-1⁺ cells treated with vehicle, ABPC (100 mg/kg), or CAM (100 mg/kg) in post-influenza pneumococcal pneumonia mice (n = 34 per group). Combined data for two independent experiments are shown. *<i>p</i> < 0.05. **<i>p</i> < 0.01 by the log-rank test. (K) Survival rate of vehicle-, ABPC-, and CAM-treated mice intranasally inoculated with recombinant IFN-γ (16 μg/kg) or PBS at 30 min and 24 h after pneumococcal infection (n = 20 per group). (L) Survival rate of vehicle- or CAM-treated WT and <i>Ifng</i>^<i>-/-</i> mice with post-influenza pneumococcal pneumonia (n = 7–16 per group).</p