Proteins significantly different between exosomes from Mtb-infected and control macrophages.
Abstract
Exosomes are generated by the multivesicular degradation of plasma membrane fusion, lysosomal, and extracellular release of intracellular vesicles. The exosome ranges from 30 to 150 nm in size. Exosomes are “bioactive vesicles” that promote intercellular communication. Exosomes contain a variety of biologically active substances packaged with proteins, lipids, and nucleic acids. After any microbe infection into the exosomes, the content of the exosomes changes and is released into the bloodstream. Such type of exosome content could be useful for basic research on exosome biology. Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (Mtb). During the Mtb infection, the exosomes played an important role in the body’s infection and immune response by releasing several exosome components providing new ideas for diagnosis, prevention, and therapeutic treatment of Mtb infection. The detection of the low abundance of the Mtb numbers or secreted peptides in the serum of TB patients is not possible. The best way of findings for diagnosis and treatment of TB could be possible by the exploration of exosome content analysis through various useful technologies. The study and analysis of exosome content would produce a road map for the future early diagnosis, prognosis estimation, efficacy monitoring, research, and application for TB.
Keywords
- exosome
- TB
- Mtb
- content
- roadmap
- serum
1. Introduction
Tuberculosis (TB) is an infectious disease caused by
Exosomes were involved in intercellular information transmission and potential medical applications. The special insight on the biological significance of the exosome is very essential for various applications in the human biological field [4]. The characterization of exosomes is very essential during immune response for a better announcement of host-pathogen interactions. Based on exosome characterization, development of various approaches would be possible to fight infections through various pathogens. When macrophages infected with the
2. Exosomes response to the Mtb
The host interactions with the pathogens are always a challenge in chronic diseases and to understand the mechanism, complexities, and sequential events. TB is a major worldwide disease and the understanding of TB immunology become a major refined since the identification of
Exosomes are the potential mediator of T cell activation. The released exosomes from mouse
EVs were packed with proteins, nucleic acids, and lipids released from the mammalian and bacterial cells. EVs played an important role through the intercellular transduction acts like a messenger. The
Proteins secreted from the Mycobacterium species were identified those were contributed to the protective immunity. Mycobacterial surface proteins were analyzed from infected macrophages. The fibronectin and 85 kDa protein complexes were identified among the mycobacterial proteins released by the infected macrophages [16].
The exosomes promoted the macrophages for the release of chemotactic factors by activating immune cells in vivo and in vitro [6]. The microvesicles and exosomes from the
3. Exosome contents and proteomic profiles of exosome proteins with TB
Exosomes are nanovesicles secreted by most but not all cells and specifically mediate intercellular communication through the transfer of genetic information of coding and noncoding RNA to recipient cells. The exosomes played an important biological role in the regulation of normal physiological and pathological processes through altered gene regulatory networks. Exosomes were targeted for the delivery of human genetic therapies through exogenous genetic cargoes such as siRNA [18].
In TB patients, the exosomes were released from the
Exosomes were loaded with the microbial proteins after
No. of proteins | Identified proteins |
---|---|
1. | 60S acidic ribosomal protein P0 |
2. | Coronin-1 C |
3. | Lupus La Protein |
4. | Heterogenous nuclear ribonucleoprotein K |
5. | Heat shock 70KDa protein 4 |
6. | Alanine -tRNA ligase, cytoplasmic |
7. | Calreticulin |
8. | Protein disulphide isomerase A3 |
9. | L-amino acid oxidase |
10. | Moesin |
11. | Nucleolin |
12. | Vimentin |
13. | Protein disulfide-isomerase A6 |
14. | Spliceosome RNA helicase DDX39B |
15. | Fermitin family homolog 3 |
16. | Programmed cell death -6 interacting protein |
17. | S-adenosylmethionine synthase isoform type -2 |
18. | Glyceral dehyde -3 phosphate dehydrogenase |
19. | ATP dependent RNA helicase A |
20. | 60 kDa heatshock protein, mitochondrial |
21. | Cytosol aminopeptidase |
22. | Ubiquitin like modifier activating enzyme-1 |
23. | ITIH4 protein |
24. | Serine/threonine protein phosphatase 2A 65kDa regulatory subunit A alpha isoform |
25. | Tryptophan t-RNA ligase cytoplasmic |
26. | Transketolase |
27. | Zyxin (fragment) |
28. | Heat shock protein HSP90-beta |
29. | Tyrosine-tRNA ligase, cytoplasmic |
30. | 6-Phosphogluconate dehydronase, decarboxylating |
31. | X-ray repair cross complementing protein-6 |
32. | 78kDa glucose regulated protein |
33. | Eukaryotic initiation factor 4A-I |
34. | Thrombospondin -4 |
35. | Bifunctional purine biosynthesis protein PURH |
36. | Staphylococcal nuclease domain containing protein-1 |
37. | Heat shock cognate 71kDa protein |
38. | Integrin beta-1 |
39. | UDP glucose 6-dehydronase |
40. | Purine nucleoside phosphorylase |
41. | Lamin B-1 |
42. | Transforming growth factor beta induced protein ig-h3 |
43. | Palmitoyl protein thioesterase -1 |
44. | Complement C4- A |
No. of proteins | Membrane associated proteins |
---|---|
1. | 60S acidic ribosomal protein P0 |
2. | Coronin-1 C |
3. | Heterogenous nuclear ribonucleoprotein K |
4. | Alanine -tRNA ligase, cytoplasmic |
5. | Calreticulin |
6. | Protein disulphide isomerase A3 |
7. | Moesin |
8. | Nucleolin |
9. | Vimentin |
10. | Protein disulfide-isomerase A6 |
11. | Fermitin family homolog 3 |
12. | Programmed cell death -6 interacting protein |
13. | Glyceral dehyde -3 phosphate dehydrogenase |
14. | ATP dependent RNA helicase A |
15. | 60 kDa heatshock protein, mitochondrial |
16. | Cytosol aminopeptidase |
17. | Serine/threonine protein phosphatase 2A 65kDa regulatory subunit A alpha isoform |
18. | Transketolase |
19. | Heat shock protein HSP90-beta |
20. | 78kDa glucose regulated protein |
21. | Eukaryotic initiation factor 4A-I |
22. | Bifunctional purine biosynthesis protein PURH |
23. | Staphylococcal nuclease domain containing protein-1 |
24. | Heat shock cognate 71 kDa protein |
25. | Integrin beta-1 |
26. | Lamin-B1 |
No. of proteins | Protein name and sequences |
---|---|
1. | Eukaryotic initiation factor 4AI and EVQkLQMEAPHIIVGTPGRVF |
2. | Glyceral dehyde 3 phosphate dehydrogenase and DNFGIVEGLMTTVHAITATQkTV |
3. | Heat shock cognate 71 kDa protein and DPVEkALR |
4. | Heat shock protein HSP90-beta and ERIMkAQALR |
5. | Moesin and EFAkEALLQASR |
6. | Nucleoside diphosphate kinase and ERTFIAIkP |
7. | Vimentin and DVRQQYESVAAkNLQEA |
A study investigated the regulation of protein N-glycosylation in human macrophages and their secreted microparticles (MPs) after
Sequence | Protein |
---|---|
a. PTB patient | |
FALNAANAR | GlcB |
VYQNAGGTHPTTTYK | Mpt64 |
AFDWDQAYR | Mpt64 |
EAPYELNITSATYQSAIPPR | Mpt64 |
b. EPTB patient | |
PGLPVEYLQVPSPSMGR | Ag85 |
FLEGLTLR | Ag85c |
LYASAEATDSK | Mpt32 |
ATIEQLLTIPLAK | GlcB |
DGQLTIK | HspX |
SEFAYGSFVR | HspX |
Sequence | Protein |
---|---|
FALNAANAR | GlcB |
DGQLTIK | HspX |
SEFAYGSFVR | HspX |
WHDPWVHASLLAQNNTR | Mpt51 |
GSVTPAVSQFNAR | Mpt63 |
VYQNAGGTHPTTTYK | Mpt64 |
EAPYELNITSATYQSAIPPR | Mpt64 |
IPDEDLAGLR | AcpM |
ATIEQLLTIPLAK | GlcB |
Exosome RNA sequencing analysis were derived from the clinical samples of ATB, LTB revealed the gene expression profiles. The selective packaging of RNA cargoes into exosomes in different stages of
These gene-enrichment analysis of the
An enhanced MRM-MS is a method to detect ultra-low abundance of ultra-
Number | Protein | Protein name | Gene |
---|---|---|---|
1 | P02671 | Fibrinogen alpha chain | FGA |
2 | G3V1N2 | HCG1745306, isoform CRA_a | HBA2 |
3 | A0N071 | Delta globin | HBD |
4 | A0A024R035 | Complement component C9 | C9 |
5 | V9HWE3 | Carbonic anhydrase | HEL-S-11 |
6 | Q9Y6C2 | EMILIN-1 | EMILIN-1 |
7 | A0A024RC61 | Aminopeptidase | ANPEP |
8 | Q8TCF0 L | Lipopolysaccharide-binding protein | LBP |
9 | P02786 | Transferrin receptor protein 1 | TFRC |
10 | P01023 | Alpha-2-macroglobulin;α-2 | A2M |
11 | C9JB55 | Serotransferrin | TF |
12 | P19652 | Alpha-1-acid glycoprotein 2 | ORM2 |
13 | Q1L857 | Ceruloplasmin | N/A |
Number | Protein | Protein name | Gene |
---|---|---|---|
1 | A3KPE2 | Apolipoprotein C-III | APOC3 |
2 | V9HVZ4 | Glyceraldehyde-3-phosphate dehydrogenase | HEL-S-162eP |
3 | B0UXD8 | HLA-DRA | HLA-DRA |
4 | P04899 | Guanine nucleotide-binding protein G(i) subunit alpha-2 | GNAI2 |
5 | E7EU05 | Glycoprotein IIIb | CD36 |
6 | P23229 | Integrin alpha-6 | ITGA6 |
7 | A0A024R4F1 | 2-phospho-D-glycerate hydro-lyase | HEL-S-17 |
8 | G8GBV0 | MHC class I antigen | HLA-A |
9 | P07737 | Profilin-1 | PFN1 |
10 | L7UUZ7 | Integrin beta | ITGB3 |
11 | Q5JP53 | Tubulin beta chain | TUBB |
12 | V9HWF0 | Integrin-linked protein kinase | HEL-S-28 |
13 | A0A024R611 | Coronin | CORO1A |
14 | V9HWN7 | Fructose-bisphosphate aldolase | HEL-S-87p |
15 | G9FP35 | Guanine nucleotide binding protein | GNAQ |
16 | D3DVF0 | Junctional adhesion molecule 1 | F11R |
17 | Q9NZN3 | EH domain-containing protein 3 | EHD3 |
18 | A0A024R3Q0 | ADP-ribosylation factor 1, isoform CRA_a | ARF1 |
19 | V9HWF5 | Peptidyl-prolyl cis-trans isomerase | HEL-S-69p |
20 | B0V023 | C6orf25 | C6orf25 |
21 | X6RJP6 | Transgelin-2 | TAGLN2 |
22 | Q12913 | Receptor-type tyrosine-protein phosphatase eta | PTPRJ |
23 | P08567 | Pleckstrin | PLEK |
24 | P48059 | LIM and senescent cell antigen-like-containing domain protein 1 | LIMS1 |
25 | Q86UX7 | Fermitin family homolog 3 | FERMT3 |
26 | Q9Y490 | Talin-1 | TLN1 |
27 | P21333 | Filamin-A | FLNA |
28 | V9HWI5 | Cofilin, non-muscle isoform | HEL-S-15 |
29 | P61160 | Actin-related protein 2 | ACTR2 |
30 | A8K0T9 | F-actin-capping protein subunit alpha | N/A |
The study and analysis of exosome contents are suitable for the development of a suitable biomarker for the diagnosis and treatment of TB. The exosome protein components were identified.
4. Exosome miRNA as a biomarker source for diagnosis and treatment of TB
Serum exosomes expressed CD81, the exosome marker protein. When these exosomes were infected with the
Now a days, TB is a threat to human health problem has an accuracy to the current TB diagnosis. Circulating exosome could be used as a diagnostic biomarker in TB. The study was examined the expression of the biomarkers for the diagnosis of TB infection. The miR-484, miR425, and miR96 were significantly increased in TB patients as compared with the healthy control and was examined the expression of miRNA as biomarker candidates for diagnosis of TB infection [31]. miRNA and electronic health records (EHRs) could be used to develop diagnostic models for PTB and tuberculosis meningitis (TBM) in a selective cohort study with the support vector machine (SVM) algorithm. Exosomal miRNAs (miR 20b, miR191 and miR486) along with EHR data through a machine learning algorithm could suggest for the diagnosis of the PTB and TBM [32]. The development of potential molecular targets for the detection and diagnosis of latent and active TB is possible by the miRNAs and repetitive region-derived small RNAs in exosomes. The most possible specifically expressed miRNA in LTBI patients were (hsa-let-7e-5p, hsa-let-7d-5p, hsa-miR-450a-5p, and hsa-miR-140-5p) and in TB patients were (hsa-miR-1246, hsa-miR-2110, hsa-miR-370-3P, hsa-miR-28-3P, and hsa-miR-193b-5p). Over all findings on miRNA, indicates the presence of biomarkers on the detection and diagnosis of the LTBI and TB patients [33].
The emerging role of functional and diagnostic potential of the several exosomal miRNA was studied by the several investigators and could explore as a possible diagnostic and therapeutic biomarker in
5. Exosomal DNA as a novel diagnostic biomarker for TB
Exosome is suitable for the detection of pathogen-derived nucleic acids. A novel approach was adopted for diagnosis of TB using exosomal DNA (exoDNA) through the droplet digital PCR (ddPCR). The ddPCR platform was used for detection of
6. Basic needs of exosomes as a biomarker content in the diagnosis and treatment of TB
Exosomes were isolated from human body fluids and considered for early detection of
7. Conclusion
Exosome emerged as a potent genetic information for therapeutic potential through transfer agents corroborating a range of biological processes. Exosomes were used as a research tool for diagnosis and treatment of TB because the exosomes were released from cells packaged with biochemical materials. The characterization and detection of various packaged biochemical materials in exosome could make a future roadmap for the diagnosis and treatment of TB in human population level.
Acknowledgments
Indian Council of Medical Research, Govt. of India is acknowledged. N S Manisha, Odisha University of Agriculture and Technology, Odisha, India is highly praised for assisting making tables and pictures in the book chapter. Thanks to Mrs. Usha Padhee, Indian Administrative Service, Principal Secretary for a supportive stand on the road map for Tuberculosis.
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