Our previous studies have shown that high alcohol-producing Klebsiella pneumoniae (HiAlc Kpn) in the intestinal microbiome could be one of the causes of non-alcoholic fatty liver disease (NAFLD). Considering antimicrobial resistance of K. pneumoniae and dysbacteriosis caused by antibiotics, phage therapy might have potential in treatment of HiAlc Kpn-induced NAFLD, because of the specificity targeting the bacteria. Here, we clarified the effectiveness of phage therapy in male mice with HiAlc Kpn-induced steatohepatitis. Comprehensive investigations including transcriptomes and metabolomes revealed that treatment with HiAlc Kpn-specific phage was able to alleviate steatohepatitis caused by HiAlc Kpn, including hepatic dysfunction and expression of cytokines and lipogenic genes. In contrast, such treatment did not cause significantly pathological changes, either in functions of liver and kidney, or in components of gut microbiota. In addition to reducing alcohol attack, phage therapy also regulated inflammation, and lipid and carbohydrate metabolism. Our data suggest that phage therapy targeting gut microbiota is an alternative to antibiotics, with potential efficacy and safety, at least in HiAlc Kpn-caused NAFLD.
Klebsiella pneumoniae
can cause a range of illnesses and has a high colonization rate in the human gut, making it crucial to develop an efficient method for detecting
K. pneumoniae
in fecal samples.
Klebsiella pneumoniae, a major pathogen leading to severe pneumonia, causes clinical complications in nosocomial and community-acquired infections. We have previously reported that high alcohol-producing K. pneumoniae (HiAlc Kpn) in the gut microbiome are strongly associated with nonalcoholic fatty liver disease (NAFLD) owing to the excess endogenous alcohol production. In the present study, we discovered that Kpn isolated from sputum and bronchoalveolar fluid (BALF) of patients with pneumoniae and chronic obstructive pulmonary disease (COPD) could also produce excess ethanol. In a murine model, HiAlc Kpn aggravated lung injury of mice with acute infection comparing with alcohol dehydrogenase gene-knockout mutant. A single-cell sequencing transcriptome analysis showed that resistin like gamma (Retnlg) neutrophil played a critical role during acute infection. The HiAlc Kpn inactivated Retnlg neutrophils in an ethanol production-dependent manner in mice, which counteract immune defense possibly through down-regulating pyrimidine and modifying DNA methylation. Furthermore, data also showed that HiAlc Kpn inhibited interleukin 1 receptor antagonist (IL1RN) gene expression by neutrophils, thereby activating IL-1β to orchestrate immune responses. Understanding functions of Retnlg neutrophils and IL-1β release in the context of HiAlc Kpn and host interactions are essential for developing novel therapeutic strategies.
Mycoplasma pneumoniae is a common causative pathogen of community-acquired pneumonia. An accurate and sensitive detection method is important for evaluating disease severity and treatment efficacy. Digital droplet PCR (ddPCR) is a competent method enabling the absolute quantification of DNA copy number with high precision and sensitivity. We established ddPCR for M. pneumoniae detection, using clinical specimens for validation, and this showed excellent specificity for M. pneumoniae. The limit of detection of ddPCR was 2.9 copies/reaction, while that for real-time PCR was 10.8 copies/reaction. In total, 178 clinical samples were used to evaluate the ddPCR assay, which correctly identified and differentiated 80 positive samples, whereas the real-time PCR tested 79 samples as positive. One sample that tested negative in real-time PCR was positive in ddPCR, with a bacterial load of three copies/test. For samples that tested positive in both methods, the cycle threshold of real-time PCR was highly correlated with the copy number of ddPCR. Bacterial loads in patients with severe M. pneumoniae pneumonia were significantly higher than those in patients with general M. pneumoniae pneumonia. The ddPCR showed that bacterial loads were significantly decreased after macrolide treatment, which could have reflected the treatment efficacy. The proposed ddPCR assay was sensitive and specific for the detection of M. pneumoniae. Quantitative monitoring of bacterial load in clinical samples could help clinicians to evaluate treatment efficacy.
High alcohol-producing
Klebsiella pneumoniae
(HiAlc
Kpn
) is one of causative agents of nonalcoholic fatty liver disease (NAFLD) and could induce liver damage. DNA methylation, as a common epigenetic form following contact with an etiologic agent and pathogenesis, can affect chromosome stability and transcription.
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