HbA_1c for Melioid and Diabetes Cohorts, and treatment received.

<p>Fig 1A shows boxplots with Tukey whiskers showing the percentage glycated haemoglobin (HbA_1c) for subjects in the <i>Melioid Cohort</i> (culture-confirmed acute melioidosis) at admission to the study with no diagnosis of Diabetes Mellitus (<i>Melioid & no DM</i>), with a new diagnosis of Diabetes Mellitus defined for the study as an HbA_1c value of ≥7% (<i>Melioid & New Diagnosis DM</i>) and with known Diabetes Mellitus (<i>Melioid & Known DM</i>) alongside subjects recruited from diabetes out-patient clinic who are (<i>DM outpatient controls</i>).**** <i>P</i> < 0.0001 by Mann-Whitney test. Fig 1B shows the drug treatment for diabetes on admission for the 117 patients in the <i>Melioid Cohort</i> with previously diagnosed diabetes mellitus.</p

Ex vivo interferon-gamma ELISPOT Responses to Heat killed Burkholderia pseudomallei by diabetes status.

<p>Ex vivo IFN-γ ELIspot responses to heat-inactivated <i>B</i>. <i>pseudomallei</i> are shown for subjects with melioidosis on admission (<i>Week 0)</i> and 12 (<i>Week12</i>) and 52 (<i>Week 52</i>) weeks later according to the presence (+) or absence (-) of a diagnosis of Diabetes Mellitus. Peripheral blood mononuclear cells (PBMC) were stimulated with heat-inactivated <i>B</i>. <i>pseudomallei</i> for 18 hours and IFN-γ secreting cells counted and expressed as spot forming cells per million PBMC (SFC/10⁶ PBMC). Subjects with diabetes had lower responses to <i>B</i>. <i>pseudomallei</i> during acute illness (Week 0) compared to subjects with no diabetes. Boxplots with Tukey whiskers are shown with responses greater than 1000 SFC/million PBMC not displayed. ** <i>P</i> < 0.01, ns = not significant by Kruskal-Wallis testing with Dunn’s multiple comparisons test.</p

Multivariable analysis for prediction of 28-day mortality in patients with acute melioidosis.

<p>The table displays the odds ratios for variables studied as predictors of mortality in 200 patients with acute melioidosis in a multivariable logistic regression model. Bp cell response is the log₁₀ transformed ELIspot response in fresh peripheral blood mononuclear cells to heat-killed <i>B</i>. <i>pseudomallei</i>. Neutrophil count represents the peripheral blood neutrophil count with > 4000 to 8000 neutrophils / μl as the comparator group. Odds ratios are adjusted for the other variables included in the model.</p><p>Multivariable analysis for prediction of 28-day mortality in patients with acute melioidosis.</p

Subject demographics.

<p>The gender, age and diabetes status of subjects in the study are shown. Two thirds (67%) of the patients with acute melioidosis were male, with a mean age of 55 years.</p><p>*Diabetes definitions are as follows: Pre-diagnosed = subjects with a previous diagnosis of diabetes mellitus by a doctor. Study definition = subjects with a previous diagnosis of diabetes mellitus by a doctor and / or having an HbA_1c of ≥ 7%.</p><p>Subject demographics.</p

A nonsense mutation in <i>TLR5</i> is associated with survival and reduced IL-10 and TNF-α levels in human melioidosis

<div><p>Background</p><p>Melioidosis, caused by the flagellated bacterium <i>Burkholderia pseudomallei</i>, is a life-threatening and increasingly recognized emerging disease. Toll-like receptor (TLR) 5 is a germline-encoded pattern recognition receptor to bacterial flagellin. We evaluated the association of a nonsense <i>TLR5</i> genetic variant that truncates the receptor with clinical outcomes and with immune responses in melioidosis.</p><p>Methodology/Principal findings</p><p>We genotyped <i>TLR5</i> c.1174C>T in 194 acute melioidosis patients in Thailand. Twenty-six (13%) were genotype CT or TT. In univariable analysis, carriage of the c.1174C>T variant was associated with lower 28-day mortality (odds ratio (OR) 0.21, 95% confidence interval (CI) 0.05–0.94, <i>P</i> = 0.04) and with lower 90-day mortality (OR 0.25, 95% CI 0.07–086, <i>P</i> = 0.03). In multivariable analysis adjusting for age, sex, diabetes and renal disease, the adjusted OR for 28-day mortality in carriers of the variant was 0.24 (95% CI 0.05–1.08, <i>P</i> = 0.06); and the adjusted OR for 90-day mortality was 0.27 (95% CI 0.08–0.97, <i>P</i> = 0.04). c.1174C>T was associated with a lower rate of bacteremia (<i>P</i> = 0.04) and reduced plasma levels of IL-10 (<i>P</i> = 0.049) and TNF-α (<i>P</i> < 0.0001). We did not find an association between c.1174C>T and IFN-γ ELISPOT (T-cell) responses (<i>P</i> = 0.49), indirect haemagglutination titers or IgG antibodies to bacterial flagellin during acute melioidosis (<i>P</i> = 0.30 and 0.1, respectively).</p><p>Conclusions/Significance</p><p>This study independently confirms the association of <i>TLR5</i> c.1174C>T with protection against death in melioidosis, identifies lower bacteremia, IL-10 and TNF-α production in carriers of the variant with melioidosis, but does not demonstrate an association of the variant with acute T-cell IFN-γ response, indirect haemagglutination antibody titer, or anti-flagellin IgG antibodies.</p></div

Patients’ characteristics and mortality.

<p>Patients’ characteristics and mortality.</p

Immune response measurements by <i>TLR5</i> c.1174C>T genotype.

<p>Immune response measurements by <i>TLR5</i> c.1174C>T genotype.</p

Kaplan-Meier survival curve of melioidosis subjects demonstrated enhanced survival in carriers of <i>TLR5</i> c.1174C>T.

<p>Curves are significantly different by the log rank test (<i>P = 0</i>.<i>03</i>).</p

Optimization of the <i>ex vivo</i> IFN-γ ELISpot assay.

<p>Optimization of the <i>ex vivo</i> IFN-γ ELISpot assay.</p

Adjusted spot counts from <i>ex-vivo</i> IFN-γ ELISpot at each time point.

<p>Adjusted spot counts from <i>ex-vivo</i> IFN-γ ELISpot at each time point.</p