Abstract
Although mycobacterial proteins in exosomes from peripheral serum of patients with tuberculosis (TB) have been identified, other exact compositions of exosomes remain unknown. In the present study, a comprehensive proteomics analysis of serum exosomes derived from patients with active TB (ATB) was performed. Exosomes from patients with ATB were characterized using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting analysis. Then identified protein components were quantified by label-free proteomics and were determined via bioinformatics analysis. A total of 123 differential proteins were identified in ATB serum exosomes and analyzed with Gene Ontology (GO) analysis. Among these proteins heat shock protein70 (HSP70), CD81, major histocompatibility complex-I (MHC-I ) and tumor susceptibility gene101 (TSG101) were present in exosomes of ATB and normal individuals confirmed via western blotting. In addition, among identified exosomal proteins lipopolysaccharide binding protein (LBP) increased significantly, but CD36 and MHC-I decreased significantly in ATB exosomes. Meanwhile, MHC-I was down-expressed in serum and peripheral blood mononuclear cells (PBMCs) of ATB, but interestingly CD36 was down-regulated in serum and up-expressed in PBMCs of ATB patients validated with ELISA and flow cytometry. CD36 was up-regulated by M. tuberculosis H37Ra infection in macrophages and suppressed in exosomes from H37Ra infected macrophages detected by western blotting. This study provided a comprehensive description of the exosome proteome in the serum of patients with ATB and revealed certain potential biomarkers associated with TB infection.
Highlights
Mycobacterium tuberculosis (M. tuberculosis), the pathogen of tuberculosis (TB), remains an important public health problem around world (Biadglegne et al, 2021)
Western blot analysis further confirmed the presence of surface markers of exosomes, such as heat shock protein70 (HSP70), CD81, and tumor susceptibility gene101 (TSG101) (Figures 1C,D)
Several markers, including HSP70 and tetraspanins (CD9, CD63, and CD81), are conserved on exosomes derived from various cells (Anand et al, 2010; Wang et al, 2014; Kowal et al, 2017)
Summary
Mycobacterium tuberculosis (M. tuberculosis), the pathogen of tuberculosis (TB), remains an important public health problem around world (Biadglegne et al, 2021). In the past 10 years, a new generation of tools, including genome sequencing and proteomics, Proteomics Analysis of Exosomes From Tuberculosis Patients have been developed to help us to further understand the disease progression of pathogens in host cells after being processed by the immune system (Sukumar et al, 2014; Cadena et al, 2016). The main techniques for diagnosing active TB are direct microscopic demonstration, culture and polymerase chain reaction (PCR) (Suárez et al, 2019). These traditional diagnostic methods are complicated and time-consuming, which hinders the rapid diagnosis of tuberculosis. It is urgent to find diagnostic markers for rapid diagnosis of tuberculosis (Goletti et al, 2016)
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