Phagocytic capacity of peripheral blood monocytes in patients with chronic liver disease (CLD).

<p>(A) Representative flow cytometry plots highlighting CD14⁺ monocyte uptake of E.Coli FITC-labelled BioParticles. The proportion of monocytes undergoing phagocytosis (B and C) and monocyte phagocytic capacity (D and E) in control subjects and CLD patients with no advanced fibrosis (No adv.), cirrhosis or decompensated cirrhosis (Decomp). (F and G) Correlation between monocyte phagocytic capacity and patient serum ALT levels. (Data represented as mean +SEM, * P<0.05, **P<0.01)</p

Peripheral blood monocyte subset distribution in chronic liver disease (CLD).

<p>(A) Representative flow cytometry plot of the 3 main monocyte subsets, i—CD14^highCD16^- “Classical”, ii—CD14^highCD16⁺ “intermediate”, iii—CD14⁺CD16⁺ “non-classical” monocytes. (B) Distribution of peripheral leukocyte lineages obtained from patient haematology full blood counts. (C) Proportion of monocytes in isolated peripheral blood mononuclear cell (PBMC) in control subjects and CLD patients with no-advanced fibrosis (No adv.), cirrhosis or decompensated cirrhosis (Decomp). (D) Distribution of monocyte subsets in the PBMC fraction of control subjects and CLD patients. (Data represented as mean +SEM, ** ^{++ ## ψψ} P<0.01; D, significance shown vs corresponding control subset; Neut, neutrophils; Lymph, lymphocytes; Mono, monocytes; Eosin, eosinophils; Baso, basophils)</p

Characterisation of monocyte phenotype in chronic liver disease (CLD).

<p>(A) Proportion of monocytes positive for common subset markers in control subjects and patients with HCV or NAFLD. (B) Level of expression (median fluorescence intensity, MFI) of selected markers on the 3 main monocyte subsets in patients with HCV. Figures are representative of subset expression in patient and control cohorts. (C) Monocyte CCR2 and HLA-DR expression in control subjects and CLD patients with no advanced fibrosis, cirrhosis or decompensated cirrhosis. (Data represented as mean +SEM, * P<0.05, **P<0.01, **P<0.001)</p

Alterations in chemokine receptor expression and adhesion molecules implicated in monocyte recruitment in patients with chronic liver disease (CLD).

<p>(A) Proportion of monocytes positive for selected chemokine receptors and adhesion molecules in control subjects and patients with HCV or NAFLD. (B) Level of expression (median fluorescence intensity, MFI) of selected markers on the 3 main monocyte subsets in patients with HCV. Figures are representative of subset expression in patient and control cohorts. (C) Expression of selected markers on monocytes from control subjects and CLD patients with no advanced fibrosis, cirrhosis or decompensated cirrhosis. (Data represented as mean +SEM, *P<0.05, **P<0.01, ***P<0.001)</p

Demographic and clinical characteristics of patient cohorts.

<p>Demographic and clinical characteristics of patient cohorts.</p

Distribution of bacterial phyla identified by 16S sequence analysis of ascites bacterial DNA.

<p>Boxes represent median values and error bars show interquartile range</p

Flow cytometry gating strategy for ascites leukocyte characterisation.

<p>(A) Lymphocytes and myeloid cells were distinguished on the basis of side scatter properties and CD14 expression. CD14Hi and CD14 Low/^negative cells were further characterised for CD16, HLA-DR and CD66B expression (top right panels). Lymphoid cells were classified as B cells (CD19+), T cells (CD3+), and NK cells (CD56+/CD16+/-) (bottom left panels). (B) Myeloid populations were further investigated for surface CD11C, CCR2, CD163 and CX3CR1 expression.</p

Ascites bacterial DNA burden associated with reduced HLA-DR expression on ascitic fluid macrophages.

<p>(A) distribution of leukocyte lineages in ascites fluid (n = 18, midline represents median, box represents 25^th-75^th percentile, whiskers indicate minimum and maximum values,-ve indicates lack of staining for markers employed in this study) (B) Correlation between %CD14+ macrophages and neutrophil count in ascitic fluid. Correlation between surface HLA-DR expression on CD14+ ascites cells (MFI) and (C) ascites bacterial burden (CFU/ml), (D) serum ascites albumin gradient (SAAG) and (E) time to next hospital admission.</p

Bacterial DNA burden in ascitic fluid is associated with poor clinical outcomes.

<p>(A) Ascitic bacterial DNA burden in patients who had received antibiotics within the previous 2 weeks (2/52) (p = 0.28). (B) Correlation between bacterial DNA burden and the number of neutrophils/ml ascitic fluid (r_s = 0.5, p = 0.012). (C) Bacterial DNA burden in patients with a previous history of SBP (p = 0.027). Correlation between bacterial DNA burden and (D) ascites total protein content (r_s = -0.42, p = 0.045) and (E) time to hospital readmission (r_s = -0.50, p = 0.024). (F) Bacterial DNA burden in patients who survived and those who died (black squares) or developed SBP (grey squares, # developed SBP and died, p = 0.006).</p

Ascites cohort antibiotic history and 6-month outcomes.

<p>Antibiotic treatment history prior to the study paracentesis and 6 month outcomes are depicted for each patient, in relation to their ascitic bacterial DNA burden (CFU/ml).</p