Glycopeptidolipid Genotype Correlates With the Severity of Mycobacterium abscessus Lung Disease.

BACKGROUND: Smooth and rough colony morphotypes of Mycobacterium abscessus are associated with virulence, but some isolates form both smooth and rough colonies, impeding successful morphotype identification. Reportedly, smooth/rough morphotypes are also related to the glycopeptidolipid (GPL) genotype. However, the accuracy of GPL genotyping to discriminate morphotypes and the relationship between GPL genotype and clinical characteristics of M abscessus lung disease have not been verified. METHODS: A retrospective analysis of colony morphology, GPL genotype, and clinical data from 182 patients with M abscessus lung disease was conducted. RESULTS: Of 194 clinical isolates, 126 (65.0%), 15 (7.7%), and 53 (27.3%) exhibited rough, smooth, and mixed colony morphotypes, respectively. Glycopeptidolipid genotyping indicated that 86.7% (13 of 15) of smooth isolates belonged to the GPL-wild type (WT) group, whereas 98.4% (124 of 126) of rough isolates belonged to the GPL-mutant type (MUT) group. Therefore, GPL genotyping accurately distinguished between smooth and rough morphotypes. Mixed colony morphotypes were also divided into GPL-WT (18.9%) and GPL-MUT (81.1%) groups. Further analysis revealed that patients infected with the GPL-MUT group presented with significantly worse baseline clinical characteristics and exacerbated episodes of lung disease. CONCLUSIONS: Glycopeptidolipid genotyping accurately distinguishes smooth and rough colony morphotypes. Patients infected with the GPL-MUT genotype exhibit worse clinical characteristics and are at a higher risk of exacerbated lung disease.

Molecular Analysis of Linezolid-Resistant Clinical Isolates of Mycobacterium abscessus.

A total of 194 Mycobacterium abscessus isolates were collected from patients, and the whole genomes were sequenced. Eighty-five (43.8%) isolates showed linezolid (LZD) resistance. Only 8.2% of resistant isolates harbored 23S rRNA mutations. Quantitative real-time PCR (qRT-PCR) revealed higher transcriptional levels of efflux pumps lmrS and mmpL9 in LZD-resistant isolates. Genome comparative analysis identified several new LZD resistance-associated genes. This study highlights the role of efflux pumps in LZD-resistant M. abscessus and proposes potential target genes for further studies.

Detection and molecular characterisation of amikacin-resistant Mycobacterium abscessus isolated from patients with pulmonary disease.

OBJECTIVES: The aim of this study was to investigate the molecular mechanisms conferring amikacin (AMK) resistance in Mycobacterium abscessus clinical isolates. METHODS: A total of 194M. abscessus clinical isolates were collected from patients with pulmonary disease during the period 2012-2017. AMK susceptibility was determined by the broth microdilution method. Whole-genome data were used for identification of mutations in resistance-associated genes. Quantitative reverse transcription PCR (qRT-PCR) was performed to measure the gene transcriptional level. RESULTS: AMK showed high in vitro killing activity against M. abscessus, with an MIC50 of 8mg/L and an MIC90 of 16mg/L. Five isolates (2.6%) were resistant to AMK (MIC>1024mg/L), of which four (80.0%) harboured a resistance-associated rrs mutation A1408G. qRT-PCR analysis showed that most of the AMK-resistant isolates (4/5; 80.0%) overexpressed the transcriptional regulator gene whiB7 and the multidrug-efflux transporter gene tap. However, overexpression of the aminoglycoside-modifying enzyme gene eis2 was only observed in one (20.0%) AMK-resistant isolate. CONCLUSION: The AMK resistance rate in M. abscessus clinical isolates in this study was low (2.6%). The A1408G mutation in rrs and overexpression of WhiB7 and Tap were the predominant mechanisms of AMK resistance in M. abscessus.

Prognostic Value of Concomitant Bronchiectasis in Newly Diagnosed Diffuse Panbronchiolitis Patients on a Maintenance Therapy with Macrolides.

Background: Factors determining the prognosis of diffuse panbronchiolitis (DPB) remain unclear at present. The objective of this study was to identify the prognostic value of concomitant bronchiectasis in the macrolide treatment efficacy and exacerbation risk in DPB patients. Methods: Data of patients initially diagnosed with DPB at the Shanghai Pulmonary Hospital between January 2007 and December 2017 were retrospectively collected and analyzed. The patients were divided into two groups according to the existence of bronchiectasis. Clinical manifestations, laboratory findings, microbiological culture results, as well as exacerbation risks and treatment outcomes, were compared between these two groups. The survival curve and Cox regression analysis models were additionally constructed to further demonstrate the predicting role of bronchiectasis in DPB exacerbation. Results: Baseline data revealed more respiratory symptoms, lower body mass index (BMI), and forced expiratory volume in one second (FEV1) as well as increased isolates of Pseudomonas aeruginosa (P. aeruginosa) in DPB subjects with bronchiectasis than those without. Furthermore, bronchiectasis was associated with a lower rate of responsiveness to macrolides and increased exacerbation frequency during follow-up. The survival curve and Cox regression analysis showed that comorbid bronchiectasis was linked to increased time to episode relapse, which remained significant even after controlling for BMI, FEV1, and P. aeruginosa culture results. Conclusion: The coexistence of bronchiectasis predicted a poor outcome of maintenance macrolide therapy and an increased exacerbation risk in DPB subjects, possibly through its impacts on nutritional status, pulmonary function, and P. aeruginosa infections.

The Serum Level of IL-1B Correlates with the Activity of Chronic Pulmonary Aspergillosis.

Background: Until now, there have been no objective criteria to determine the activity of chronic pulmonary aspergillosis (CPA). This study aims to analyze the correlation between serum level of IL-1B and the activity of CPA and to determine whether serum IL-1B could be used to assess the activity of CPA. Methods: A total of 469 newly diagnosed CPA patients were enrolled. Correlation analysis in the whole subjects showed that only IL-1B level was associated with the activity of CPA. Then, 381 cases with factors significantly affecting IL-1B expression was excluded through multiple linear regression; the remaining 88 patients were divided into high IL-1B group and low IL-1B group, according to the median value of serum IL-1B, for subgroup analysis. A retrospective comparative analysis was subsequently performed between the two groups, including the clinical manifestation, microbiology and laboratory tests results, and imaging findings. We further investigated the relationship between IL-1B levels and CT characteristic which acted as the indicator of CPA activity, as well as changes in IL-1B level before and after surgery. Results: For all patients, correlation analysis revealed that IL-1B level correlated with both cavitary diameter (P=0.035) and aspergilloma size (P<0.047) but not with the thickness of the cavity (P=0.479). In subgroup comparative analysis, CT characteristics suggested that high activity of CPA, such as cavitary (27/44 vs 13/44, P=0.003) and aspergilloma lesions (25/44 vs. 11/44, P<0.002), were more frequently found in high IL-1B group. The cavity diameter (P<0.001), aspergilloma size (P=0.006), and cavity wall thickness (P=0.023) were significantly different between the two groups. When Spearman correlation analysis was performed once again in subgroup, an even stronger relationship of serum IL-1B with the cavity diameter (Rs=0.501, P=0.002) and aspergilloma size (Rs=0.615, P=0.001) was observed. Interestingly, a significant reduction of IL-1B level was observed after successful resection of CPA lesions. Conclusion: Higher level of serum IL-1B is associated with more severe cavitary and aspergilloma lesions, which are indicative of more active CPA. In addition, IL-1B level reduced accordingly after lesion resection. Measuring IL-1B level therefore could be served as a convenient method to monitor the activity of CPA and be a potential predictive/prognostic marker for treatment response.