1887

Journal of Medical Microbiology logo

Volume 69, Issue 10

An increase in p62/NBR1 levels in melioidosis patients of Sri Lanka exhibit a characteristic of potential host biomarker Open Access

Abstract

Melioidosis, caused by , in endemic areas, poses a challenge for treating the diseased populations without accurate diagnosis, and the disease-specific biomarkers linked with the infection have yet to be reported. Due to the invasive nature of the causative agent, , host innate effector mechanisms, including autophagy are known to be activated, resulting in differential expression of cellular proteins and immune markers. Identification of a disease-specific biomarker associated with infection will be helpful to facilitate rapid confirmation of melioidosis, which would enable early treatment and therapeutic success.

We aimed to assess the levels of a host autophagy component, p62/NBR1, which function as a cargo-receptor in the process of autophagy activation leading to the degradation of ubiquitin-coated intracellular bacteria in which p62/NBR1 itself is degraded in the clearance of the pathogen. We further probed the extent of intracellular p62/NBR1 degradation and assessed its potential as a melioidosis biomarker.

We analysed peripheral blood mononuclear cell (PBMC) lysates using an ELISA-based assay for detecting cytosolic autophagy-related proteins p62/NBR1. We measured p62/NBR1 levels in diseased (confirmed infection) and non -diseased populations and utilized receiver operating characteristic (ROC) curve and max Youden index analysis for evaluating potential disease biomarker characteristics.

Our results revealed a three to fivefold increase in p62/NBR1 levels confirmed melioidosis cases compared to uninfected healthy donors. Comparable to p62/NBR1, levels of cytosolic LC3-I levels also increased, whereas the levels of degraded membrane bound form LC3-II was low, suggesting autophagy deficiency. Proinflammatory serum cytokine response, particularly IL-6, was consistently higher alongside infection in comparison to healthy controls.

ROC curve and max Youden index analysis suggest that increased p62/NBR1 levels in diseased populations display characteristics of a potential disease biomarker in melioidosis and illustrates that an elevated p62/NBR1 level, in conjunction with infection associated with autophagy deficiency.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): autophagy , Burkholderia pseudomallei , cytokines , melioidosis , NBR1 and p62
Funding
This study was supported by the:
  • U.S. Army Medical Research Acquisition Activity (Award (USAMRAA) contract number (W81XWH-14-C-0071)
    • Principle Award Recipient: Aruna Dharshan De Silva
  • Defense Threat Reduction Agency (Award JSTO (CB3948) under USAMRIID project number 1324392)
    • Principle Award Recipient: Kamal Saikh
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001242
2020-08-20
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/jmm/69/10/1240.html?itemId=/content/journal/jmm/10.1099/jmm.0.001242&mimeType=html&fmt=ahah

References

  1. Limmathurotsakul D, Peacock SJ. Melioidosis: a clinical overview. Br Med Bull 2011; 99:125–139 [View Article][PubMed]
     [Google Scholar]
  2. Wiersinga WJ, Currie BJ, Peacock SJ. Melioidosis. N Engl J Med 2012; 367:1035–1044 [View Article][PubMed]
     [Google Scholar]
  3. Currie BJ, Ward L, Cheng AC. The epidemiology and clinical spectrum of melioidosis: 540 cases from the 20 year Darwin prospective study. PLoS Negl Trop Dis 2010; 4:e900 [View Article][PubMed]
     [Google Scholar]
  4. Rammaert B, Goyet S, Tarantola A. Melioidosis requires better data sharing for improved diagnosis and management in the Mekong region. Am J Trop Med Hyg 2014; 90:383 [View Article][PubMed]
     [Google Scholar]
  5. Limmathurotsakul D, Wongratanacheewin S, Teerawattanasook N, Wongsuvan G, Chaisuksant S et al. Increasing incidence of human melioidosis in northeast Thailand. Am J Trop Med Hyg 2010; 82:1113–1117 [View Article][PubMed]
     [Google Scholar]
  6. Corea EM, de Silva AD, Thevanesam V. Melioidosis in Sri Lanka. Trop Med Infect Dis 2018; 3:E22 [View Article][PubMed]
     [Google Scholar]
  7. Cheng AC, Currie BJ. Melioidosis: epidemiology, pathophysiology, and management. Clin Microbiol Rev 2005; 18:383–416 [View Article][PubMed]
     [Google Scholar]
  8. CDC possession, use, and transfer of select agents and toxins; biennial review Final rule. Fed, regist 2012; 77:61083–61115
     [Google Scholar]
  9. Stone JK, DeShazer D, Brett PJ, Burtnick MN. Melioidosis: molecular aspects of pathogenesis. Expert Rev Anti Infect Ther 2014; 12:1487–1499 [View Article][PubMed]
     [Google Scholar]
  10. Ooi WF, Ong C, Nandi T, Kreisberg JF, Chua HH et al. The condition-dependent transcriptional landscape of Burkholderia pseudomallei . PLoS Genet 2013; 9:e1003795 [View Article][PubMed]
     [Google Scholar]
  11. Pruksachartvuthi S, Aswapokee N, Thankerngpol K. Survival of Pseudomonas pseudomallei in human phagocytes. J Med Microbiol 1990; 31:109–114 [View Article][PubMed]
     [Google Scholar]
  12. Jones AL, Beveridge TJ, Woods DE. Intracellular survival of Burkholderia pseudomallei . Infect Immun 1996; 64:782–790 [View Article][PubMed]
     [Google Scholar]
  13. Harley VS, Dance DA, Tovey G et al. An ultrastructural study of the phagocytosis of Burkhlderia pseudomelli . Microbios 1998; 94:34–35
     [Google Scholar]
  14. Kespichayawattana W, Rattanachetkul S, Wanun T, Utaisincharoen P, Sirisinha S et al. Burkholderia pseudomallei induces cell fusion and actin-associated membrane protrusion: a possible mechanism for cell-to-cell spreading. Infect Immun 2000; 68:5377–5384 [View Article][PubMed]
     [Google Scholar]
  15. Harley VS, Dance DA, Tovey G, Drasar BS. Interaction of Pseudomonas pseudomallei with macrophages. Biochem Soc Trans 1994; 22:88S [View Article][PubMed]
     [Google Scholar]
  16. Harley VS, Dance DA, Tovey G et al. An ultrastructural study of the phagocytosis of Burkhlderia pseudomelli . Microbios 1998; 94:5–6
     [Google Scholar]
  17. Nathan SA, Puthucheary SD. An electronmicroscopic study of the interaction of Burkholderia pseudomallei and human macrophages. Malays J Pathol 2005; 27:3–7[PubMed]
     [Google Scholar]
  18. Prevatt AL, Hunt JS. Chronic systemic melioidosis; review of literature and report of a case, with a note on visual disturbance due to chloramphenicol. Am J Med 1957; 23:810–823 [View Article][PubMed]
     [Google Scholar]
  19. Newland RC. Chronic melioidosis: a case in Sydney. Pathology 1969; 1:149–152 [View Article][PubMed]
     [Google Scholar]
  20. Chodimella U, Hoppes WL, Whalen S, Ognibene AJ, Rutecki GW et al. Septicemia and suppuration in a Vietnam veteran. Hosp Pract 1997; 32:219–221 [View Article]
     [Google Scholar]
  21. Gee JE, Gulvik CA, Elrod MG, Batra D, Rowe LA et al. Phylogeography of Burkholderia pseudomallei isolates, Western hemisphere. Emerg Infect Dis 2017; 23:1133–1138 [View Article][PubMed]
     [Google Scholar]
  22. Mizushima N, Klionsky DJ. Protein turnover via autophagy: implications for metabolism. Annu Rev Nutr 2007; 27:19–40 [View Article]
     [Google Scholar]
  23. Kirkegaard K, Taylor MP, Jackson WT. Cellular autophagy: surrender, avoidance and subversion by microorganisms. Nat Rev Microbiol 2004; 2:301–314 [View Article]
     [Google Scholar]
  24. Levine B. Eating oneself and uninvited guests: autophagy-related pathways in cellular defense. Cell 2005; 120:159–162 [View Article][PubMed]
     [Google Scholar]
  25. Kirkin V, Lamark T, Sou Y-S, Bjørkøy G, Nunn JL et al. A role for NBR1 in autophagosomal degradation of ubiquitinated substrates. Mol Cell 2009; 33:505–516 [View Article][PubMed]
     [Google Scholar]
  26. Lazar Adler NR, Govan B, Cullinane M, Harper M, Adler B et al. The molecular and cellular basis of pathogenesis in melioidosis: how does Burkholderia pseudomallei cause disease?. FEMS Microbiol Rev 2009; 33:1079–1099 [View Article][PubMed]
     [Google Scholar]
  27. Saikh KU, Dankmeyer JL, Zeng X, Ulrich RG, Amemiya K et al. An increase in intracellular p62/NBR1 and persistence of Burkholderia mallei and B. pseudomallei in infected mice linked to autophagy deficiency. Immun Inflamm Dis 2019; 7:7–21 [View Article][PubMed]
     [Google Scholar]
  28. Natesan M, Corea E, Krishnananthasivam S, Sathkumara HD, Dankmeyer JL et al. Calprotectin as a biomarker for melioidosis disease progression and management. J Clin Microbiol 2017; 55:1205–1210 [View Article][PubMed]
     [Google Scholar]
  29. Patel N, Conejero L, De Reynal M, Easton A, Bancroft GJ et al. Development of vaccines against Burkholderia pseudomallei . Front Microbiol 2011; 2:198 [View Article][PubMed]
     [Google Scholar]
  30. Krishnananthasivam S, Sathkumara HD, Corea E, Natesan M, De Silva AD et al. Gene expression profile of human cytokines in response to Burkholderia pseudomallei infection. mSphere 2017; 2:e00121–17 [View Article][PubMed]
     [Google Scholar]
  31. Krishnananthasivam S, Jayathilaka N, Sathkumara HD, Corea E, Natesan M et al. Host gene expression analysis in Sri Lankan melioidosis patients. PLoS Negl Trop Dis 2017; 11:e0005643 [View Article][PubMed]
     [Google Scholar]
  32. Cullinane M, Gong L, Li X, Lazar-Adler N, Tra T et al. Stimulation of autophagy suppresses the intracellular survival of Burkholderia pseudomallei in mammalian cell lines. Autophagy 2008; 4:744–753 [View Article][PubMed]
     [Google Scholar]
  33. Wiersinga WJ, Virk HS, Torres AG, Currie BJ, Peacock SJ et al. Melioidosis. Nat Rev Dis Primers 2018; 4:17107 [View Article][PubMed]
     [Google Scholar]
  34. Skvarc M, Stubljar D, Rogina P, Kaasch AJ. Non-culture-based methods to diagnose bloodstream infection: does it work?. Eur J Microbiol Immunol 2013; 3:97–104 [View Article][PubMed]
     [Google Scholar]
  35. Friedland JS, Suputtamongkol Y, Remick DG, Chaowagul W, Strieter RM et al. Prolonged elevation of interleukin-8 and interleukin-6 concentrations in plasma and of leukocyte interleukin-8 mRNA levels during septicemic and localized Pseudomonas pseudomallei infection. Infect Immun 1992; 60:2402–2408 [View Article][PubMed]
     [Google Scholar]
  36. Suckling CJ, Alam S, Olson MA, Saikh KU, Harnett MM et al. Small molecule analogues of the parasitic worm product ES-62 interact with the TIR domain of MyD88 to inhibit pro-inflammatory signalling. Sci Rep 2018; 8:2123 [View Article][PubMed]
     [Google Scholar]
  37. Foundation for Statistical Computing Vienna, Austria. https://www.R-project.org/.
  38. Dinno A. dunn.test: Dunn's Test of Multiple Comparisons Using Rank Sums. R package version 1.3.5. https://CRAN.R-project.org/package=dunn.test ; 2017
  39. SAS Institute Inc SAS online Doc 9.4 Cary, NC; 2013
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001242
Loading
An increase in p62/NBR1 levels in melioidosis patients of Sri Lanka exhibit a characteristic of potential host biomarker
J. Med. Microbiol. 69, 1240 (2020); https://doi.org/10.1099/jmm.0.001242
/content/journal/jmm/10.1099/jmm.0.001242
/content/journal/jmm/10.1099/jmm.0.001242
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error