Introduction Patients with alcohol-related cirrhosis (ARC) are particularly prone to infection which is frequently a precipitant of organ failure and death. Neutrophil dysfunction has been reported in patients with ARC. Sepsis and associated endotoxemia occur in approximately 40% of hospitalised patients. Neutrophil TLRs are able to sense pathogens and induce inflammatory responses but whether active alcohol drinking is protective or detrimental in this context remains unknown. The aim of this study was to characterise the neutrophil phenotype, TLR2/4/9 expression and plasma cytokine profile in patients with ARC who were abstinent (n=13) compared to those who were drinking (n=16), split by MELD score15 compared to healthy controls (n=12).
Methods Neutrophils isolated from patients with ARC were studied ex vivo at baseline, and following 2-h stimulation with lipopolysaccharide (LPS) and the bacterial degradation protein fMLP. Neutrophil phenotype and TLR expression were determined using anti-CD16 (PE); -CD11b (APC-Cy7); -TLR2 (Alexa Fluor 488); -TLR4 (biotin conjugated PE-Cy7 Streptavidin) and -TLR9 (APC) by flow cytometry. Intracellular cytokine production pre- and post-stimulation will be measured by CBA.
Results The severity of cirrhosis (MELD15) or abstinent/active drinking status did not significantly impact on resting CD16, CD11b, TLR2 and 9 expression. TLR 2/9 expression on exposure to LPS and fMLP showed downregulation in those who were active alcohol-drinkers compared to the baseline. Conversely TLR4 expression was upregulated in the abstinent group with MELD>15; an effect significantly abrogated by the effect of active drinking (p=0.004). Active alcohol consumption did not however change TLR4 expression in those with MELD.
Conclusion Active alcohol consumption was shown to downregulate functional TLR2/9 expression resulting in immunoparesis with potential susceptibility to Gram-positive infection. However, active alcohol consumption was shown to abrogate the increased TLR4 responses to endotoxin stimulation seen in the abstinent ARC cohort with MELD >15. This implies that alcohol paradoxically promotes endotoxin tolerance and may act as a potential protective mechanism against Gram-negative insults in those with advanced cirrhosis.
Competing interests None declared.
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