Serum Small Extracellular Vesicles Proteome of Tuberculosis Patients Demonstrated Deregulated Immune Response

Abstract

Purpose Detailed understanding of host pathogen interaction in tuberculosis is an important avenue for identifying novel therapeutic targets. Small extracellular vesicles (EVs) like exosomes that are rich in proteins, nucleic acids and lipids, act as messengers and may show altered composition in disease conditions. Experimental design In this case control study, small EVs are isolated from serum of 58 subjects (all male, 33 (15–70) in years) including drug naïve active tuberculosis (ATB: n = 22), non‐tuberculosis (NTB: n = 18), and healthy subjects (n = 18). Serum small EVs proteome analysis is carried out using isobaric tag for relative and absolute quantification (iTRAQ) experiments and an independent sample (n = 36) is used for validation. Results A set of 132 and 68 proteins are identified in iTRAQ‐I (ATB/Healthy) and iTRAQ‐II (ATB/NTB) experiments, respectively. Four proteins (KYAT3, SERPINA1, HP, and APOC3) show deregulation (log2‐fold change > ±0.48, p < 0.05) in ATB with respect to healthy controls and Western blot data corroborated mass spectrometry findings. Conclusions and clinical relevance These important proteins, involved in neutrophil degranulation, plasma heme scavenging, kynurenine, and lipid metabolism, show deregulation in ATB patients. Identification of such a protein panel in circulating small EVs besides providing novel insights into their role in tuberculosis may prove to be useful targets to develop host‐directed therapeutic intervention.

Serum proteomics of active tuberculosis patients and contacts reveals unique processes activated during Mycobacterium tuberculosis infection

“…Using MALDI-TOF mass spectrometry (no tryptic digestion and no fragmentation) Zhang et al 33 found 236 serum "protein peaks" differentially expressed between patients with Intestinal Tuberculosis (ITB) and Crohn´s Disease (CD). In another study by Arya et al 34 , 132 and 68 serum extracellular vesicle (EVs) proteins from active TB patients were identified when compare to non-tuberculosis and healthy patients. The number of identified proteins were higher (more than seven hundred with at least one unique peptide) in the study by Chen et al 35 by analyzing immune-depleted serum samples.…”

Extracellular Vesicles in Blood: Sources, Effects, and Applications

“…Mycobacterium tuberculosis peptides have been found in circulating EVs and, moreover, the use of highly sensitive methods have enabled the detection of peptides at extremely low abundance, which could help disease diagnosis over the current sputum sample analyses [ 138 ]. Apart from exogenous molecules, a recent work reported that there are altered levels of host proteins in serum EVs from active tuberculosis patients, which could be linked to an altered immune system functioning [ 139 ]. Similarly, the characterization of plasma EVs from HIV patients have shown significant alterations.…”

Monocyte-related gene biomarkers for latent and active tuberculosis

“…Serine protease inhibitor A1 (SERPINA1) has a highly conserved protein structure, and its proportion contributes to more than 90% of the total plasma protease inhibitory ability. Arya et al [ 38 ] showed that SERPINA1 was highly expressed in active tuberculosis through the iTRAQ-I experiment, which was confirmed by Western blotting. As an acute-phase reactive protein, SERPINA1 is primarily synthesized in liver cells, but can also be produced by epithelial cells, monocytes, and macrophages.…”