Sputum Proteomics Reveals a Shift in Vitamin D-binding Protein and Antimicrobial Protein Axis in Tuberculosis Patients

Abstract

Existing understanding of molecular composition of sputum and its role in tuberculosis patients is variously limited to its diagnostic potential. We sought to identify infection induced sputum proteome alteration in active/non tuberculosis patients (A/NTB) and their role in altered lung patho-physiology. Out of the study population (n = 118), sputum proteins isolated from discovery set samples (n = 20) was used for an 8-plex isobaric tag for relative and absolute concentration analysis. A minimum set of protein with at least log2(ATB/NTB) >±1.0 in ATB was selected as biosignature and validated in 32 samples. Predictive accuracy was calculated from area under the receiver operating characteristic curve (AUC of ROC) using a confirmatory set (n = 50) by Western blot analysis. Mass spectrometry analysis identified a set of 192 sputum proteins, out of which a signature of β-integrin, vitamin D binding protein:DBP, uteroglobin, profilin and cathelicidin antimicrobial peptide was sufficient to differentiate ATB from NTB. AUC of ROC of the biosignature was calculated to 0.75. A shift in DBP-antimicrobial peptide (AMP) axis in the lungs of tuberculosis patients is observed. The identified sputum protein signature is a promising panel to differentiate ATB from NTB groups and suggest a deregulated DBP-AMP axis in lungs of tuberculosis patients.

A One Health Approach to Simple Liquid Sample Handling for Respiratory Based -Omics Analysis; Tracheal Wash and Bronchoalveolar Lavage Fluid

“…Proteomic analysis revealed a wide range of proteins in the equine TW samples, many of which have already been detected and reported in previously published studies on human and equine BALF and human sputum 30,[32][33][34] . Some of the detected proteins have previously been proposed as potential biomarkers for human lung cancer; these include carbonic anhydrase 1 (CA1), apolipoprotein A2 (APOA2), transthyretin (TTR), clusterin CLU, ceruloplasmin (CP) and gelsolin (GSN) 33 .…”

Decontamination of SARS-CoV-2 and other RNA viruses from N95 level meltblown polypropylene fabric using heat under different humidities

“…We next aimed to determine conditions of heat treatment and humidity that are efficient in inactivating SARS-CoV-2 loaded on the meltblown fabric. We mixed our virus stock 1:10 with either phosphate buffered saline (PBS) or bovine serum albumin (BSA) to a final concentration of 3g/L, to mimic the conditions of bodily fluids such as sputum, 28-30 which contain higher protein concentrations than is found in the media used to create virus stocks; this is relevant because high protein concentrations are known to stabilize viruses, including SARS-CoV-2. 31 We allowed droplets containing SARS-CoV-2 (backtiter 1 × 10 6 PFU/sample) dry on top of a piece of meltblown fabric in a biosafety cabinet for 2 hours.…”

Perspectives for systems biology in the management of tuberculosis

“…In a cohort of children with symptoms compatible with tuberculosis, a three-protein signature distinguished between tuberculosis, irrespective of microbiological confirmation, and other respiratory diseases with a sensitivity of 72.2% and a specificity of 75% [83]. Sputum proteomics identified a five-protein signature differentiating between tuberculosis and nontuberculosis patients (sensitivity 70%, specificity 79%) [84]. Differential regulation of pro-inflammatory, lipid and iron transport proteins were revealed between patients with active tuberculosis and LTBI patients [74].…”