Skip to main content
Top
Published in: BMC Immunology 1/2021

Open Access 01-12-2021 | Tuberculosis | Research

Ex vivo mRNA expression of toll-like receptors during latent tuberculosis infection

Authors: Birhan Alemnew, Soren T. Hoff, Tamrat Abebe, Markos Abebe, Abraham Aseffa, Rawleigh Howe, Liya Wassie

Published in: BMC Immunology | Issue 1/2021

Login to get access

Abstract

Background

Understanding immune mechanisms, particularly the role of innate immune markers during latent TB infection remains elusive. The main objective of this study was to evaluate mRNA gene expression patterns of toll-like receptors (TLRs) as correlates of immunity during latent TB infection and further infer their roles as potential diagnostic biomarkers.

Methods

Messenger RNA (mRNA) levels were analysed in a total of 64 samples collected from apparently healthy children and adolescents latently infected with tuberculosis (n = 32) or non-infected (n = 32). Relative expression in peripheral blood of selected genes encoding TLRs (TLR-1, TLR-2, TLR-4, TLR-6 and TLR-9) was determined with a quantitative real-time polymerase chain reaction (qRT-PCR) using specific primers and florescent labelled probes and a comparative threshold cycle method to define fold change. Data were analysed using Graph-Pad Prism 7.01 for Windows and a p-value less than 0.05 was considered statistically significant.

Results

An increased mean fold change in the relative expression of TLR-2 and TLR-6 mRNA was observed in LTBI groups relative to non-LTBI groups (p < 0.05), whereas a slight fold decrease was observed for TLR-1 gene.

Conclusions

An increased mRNA expression of TLR-2 and TLR-6 was observed in latently infected individuals relative to those non-infected, possibly indicating the roles these biomarkers play in sustenance of the steady state interaction between the dormant TB bacilli and host immunity.
Appendix
Available only for authorised users
Literature
1.
go back to reference World Health Organization. Global tuberculosis report 2018. Geneva: World Health Organization, 2018. World Health Organization. Global tuberculosis report 2018. Geneva: World Health Organization, 2018.
3.
go back to reference Chaves AS, Rodrigues MF, Mattos AM, Teixeira HC. Challenging mycobacterium tuberculosis dormancy mechanisms and their immunodiagnostic potential. Braz J Infect Dis. 2015;19(6):636–42.PubMedCrossRef Chaves AS, Rodrigues MF, Mattos AM, Teixeira HC. Challenging mycobacterium tuberculosis dormancy mechanisms and their immunodiagnostic potential. Braz J Infect Dis. 2015;19(6):636–42.PubMedCrossRef
5.
6.
go back to reference Walzl G, Ronacher K, Hanekom W, Scriba TJ, Zumla A. Immunological biomarkers of tuberculosis. Nat Rev Immunol. 2011;11(5):343–54.PubMedCrossRef Walzl G, Ronacher K, Hanekom W, Scriba TJ, Zumla A. Immunological biomarkers of tuberculosis. Nat Rev Immunol. 2011;11(5):343–54.PubMedCrossRef
7.
go back to reference Kleinnijenhuis J, van Crevel R, Netea MG. Trained immunity: consequences for the heterologous effects of BCG vaccination. Trans R Soc Trop Med Hyg. 2015;109(1):29–35.PubMedCrossRef Kleinnijenhuis J, van Crevel R, Netea MG. Trained immunity: consequences for the heterologous effects of BCG vaccination. Trans R Soc Trop Med Hyg. 2015;109(1):29–35.PubMedCrossRef
8.
go back to reference Yanez A, Hassanzadeh-Kiabi N, Ng MY, Megias J, Subramanian A, Liu GY, et al. Detection of a TLR2 agonist by hematopoietic stem and progenitor cells impacts the function of the macrophages they produce. Eur J Immunol. 2013;43(8):2114–25.PubMedCrossRefPubMedCentral Yanez A, Hassanzadeh-Kiabi N, Ng MY, Megias J, Subramanian A, Liu GY, et al. Detection of a TLR2 agonist by hematopoietic stem and progenitor cells impacts the function of the macrophages they produce. Eur J Immunol. 2013;43(8):2114–25.PubMedCrossRefPubMedCentral
9.
go back to reference Dowling JK, Mansell A. Toll-like receptors: the swiss army knife of immunity and vaccine development. Clin Transl Immunol. 2016;5(5):e85.CrossRef Dowling JK, Mansell A. Toll-like receptors: the swiss army knife of immunity and vaccine development. Clin Transl Immunol. 2016;5(5):e85.CrossRef
10.
go back to reference Skevaki C, Pararas M, Kostelidou K, Tsakris A, Routsias JG. Single nucleotide polymorphisms of toll-like receptors and susceptibility to infectious diseases. Clin Exp Immunol. 2015;180(2):165–77.PubMedCrossRefPubMedCentral Skevaki C, Pararas M, Kostelidou K, Tsakris A, Routsias JG. Single nucleotide polymorphisms of toll-like receptors and susceptibility to infectious diseases. Clin Exp Immunol. 2015;180(2):165–77.PubMedCrossRefPubMedCentral
11.
go back to reference Dorhoi A, Kaufmann SH. Pathology and immune reactivity: understanding multidimensionality in pulmonary tuberculosis. Semin Immunopathol. 2016;38(2):153–66.PubMedCrossRef Dorhoi A, Kaufmann SH. Pathology and immune reactivity: understanding multidimensionality in pulmonary tuberculosis. Semin Immunopathol. 2016;38(2):153–66.PubMedCrossRef
13.
go back to reference Wassie L, Aseffa A, Abebe M, Gebeyehu MZ, Zewdie M, Mihret A, et al. Parasitic infection may be associated with discordant responses to QuantiFERON and tuberculin skin test in apparently healthy children and adolescents in a tuberculosis endemic setting, Ethiopia. BMC infect Dis. 2013;13:265.PubMedCrossRefPubMedCentral Wassie L, Aseffa A, Abebe M, Gebeyehu MZ, Zewdie M, Mihret A, et al. Parasitic infection may be associated with discordant responses to QuantiFERON and tuberculin skin test in apparently healthy children and adolescents in a tuberculosis endemic setting, Ethiopia. BMC infect Dis. 2013;13:265.PubMedCrossRefPubMedCentral
14.
go back to reference American Thoracic Society. Targeted tuberculin testing and treatment of latent tuberculosis infection. MMWR Morb Mort Wkly Rep. 2000;49(RR-6):1–51. American Thoracic Society. Targeted tuberculin testing and treatment of latent tuberculosis infection. MMWR Morb Mort Wkly Rep. 2000;49(RR-6):1–51.
15.
go back to reference EDHS. Ethiopia Demographic and Health Survey 2016. 2016. EDHS. Ethiopia Demographic and Health Survey 2016. 2016.
16.
go back to reference Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 2001;25(4):402–8.PubMedCrossRef Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 2001;25(4):402–8.PubMedCrossRef
17.
go back to reference Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ(−delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3(3):71–85.PubMedPubMedCentral Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ(−delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3(3):71–85.PubMedPubMedCentral
18.
go back to reference Zellweger JP, Sotgiu G, Corradi M, Durando P. The diagnosis of latent tuberculosis infection (LTBI): currently available tests, future developments, and perspectives to eliminate tuberculosis (TB). Med Lav. 2020;111(3):170–83.PubMedPubMedCentral Zellweger JP, Sotgiu G, Corradi M, Durando P. The diagnosis of latent tuberculosis infection (LTBI): currently available tests, future developments, and perspectives to eliminate tuberculosis (TB). Med Lav. 2020;111(3):170–83.PubMedPubMedCentral
19.
go back to reference Della Bella C, Spinicci M, Alnwaisri HFM, Bartalesi F, Tapinassi S, Mencarini J, et al. LIOFeron(R)TB/LTBI: a novel and reliable test for LTBI and tuberculosis. Int J Infect Dis. 2020;91:177–81.PubMedCrossRef Della Bella C, Spinicci M, Alnwaisri HFM, Bartalesi F, Tapinassi S, Mencarini J, et al. LIOFeron(R)TB/LTBI: a novel and reliable test for LTBI and tuberculosis. Int J Infect Dis. 2020;91:177–81.PubMedCrossRef
20.
go back to reference Lerm M, Netea MG. Trained immunity: a new avenue for tuberculosis vaccine development. J Intern Med. 2016;279(4):337–46.PubMedCrossRef Lerm M, Netea MG. Trained immunity: a new avenue for tuberculosis vaccine development. J Intern Med. 2016;279(4):337–46.PubMedCrossRef
22.
go back to reference Thada S, Valluri VL, Gaddam SL. Influence of toll-like receptor gene polymorphisms to tuberculosis susceptibility in humans. Scand J Immunol. 2013;78(3):221–9.PubMedCrossRef Thada S, Valluri VL, Gaddam SL. Influence of toll-like receptor gene polymorphisms to tuberculosis susceptibility in humans. Scand J Immunol. 2013;78(3):221–9.PubMedCrossRef
23.
go back to reference Wu L, Hu Y, Li D, Jiang W, Xu B. Screening toll-like receptor markers to predict latent tuberculosis infection and subsequent tuberculosis disease in a Chinese population. BMC Med Genet. 2015;16:19.PubMedCrossRefPubMedCentral Wu L, Hu Y, Li D, Jiang W, Xu B. Screening toll-like receptor markers to predict latent tuberculosis infection and subsequent tuberculosis disease in a Chinese population. BMC Med Genet. 2015;16:19.PubMedCrossRefPubMedCentral
24.
go back to reference Biyikli OO, Baysak A, Ece G, Oz AT, Ozhan MH, Berdeli A. Role of toll-like receptors in tuberculosis infection. Jundishapur J Microbiol. 2016;9(10):e20224.PubMedCrossRefPubMedCentral Biyikli OO, Baysak A, Ece G, Oz AT, Ozhan MH, Berdeli A. Role of toll-like receptors in tuberculosis infection. Jundishapur J Microbiol. 2016;9(10):e20224.PubMedCrossRefPubMedCentral
25.
go back to reference Faridgohar M, Nikoueinejad H. New findings of toll-like receptors involved in mycobacterium tuberculosis infection. Pathog Glob Health. 2017;111(5):256–64.PubMedCrossRefPubMedCentral Faridgohar M, Nikoueinejad H. New findings of toll-like receptors involved in mycobacterium tuberculosis infection. Pathog Glob Health. 2017;111(5):256–64.PubMedCrossRefPubMedCentral
26.
go back to reference Zhao L, Liu K, Kong X, Tao Z, Wang Y, Liu Y. Association of polymorphisms in toll-like receptors 4 and 9 with risk of pulmonary tuberculosis: a meta-analysis. Med Sci Monit. 2015;21:1097–106.PubMedCrossRefPubMedCentral Zhao L, Liu K, Kong X, Tao Z, Wang Y, Liu Y. Association of polymorphisms in toll-like receptors 4 and 9 with risk of pulmonary tuberculosis: a meta-analysis. Med Sci Monit. 2015;21:1097–106.PubMedCrossRefPubMedCentral
27.
go back to reference Constantoulakis P, Filiou E, Rovina N, Chras G, Hamhougia A, Karabela S, et al. In vivo expression of innate immunity markers in patients with mycobacterium tuberculosis infection. BMC Infect Dis. 2010;10:243.PubMedCrossRefPubMedCentral Constantoulakis P, Filiou E, Rovina N, Chras G, Hamhougia A, Karabela S, et al. In vivo expression of innate immunity markers in patients with mycobacterium tuberculosis infection. BMC Infect Dis. 2010;10:243.PubMedCrossRefPubMedCentral
28.
go back to reference Bryson BD, Rosebrock TR, Tafesse FG, Itoh CY, Nibasumba A, Babunovic GH, et al. Heterogeneous GM-CSF signaling in macrophages is associated with control of mycobacterium tuberculosis. Nat Commun. 2019;10(1):2329.PubMedCrossRefPubMedCentral Bryson BD, Rosebrock TR, Tafesse FG, Itoh CY, Nibasumba A, Babunovic GH, et al. Heterogeneous GM-CSF signaling in macrophages is associated with control of mycobacterium tuberculosis. Nat Commun. 2019;10(1):2329.PubMedCrossRefPubMedCentral
29.
go back to reference Romieu-Mourez R, Francois M, Boivin MN, Bouchentouf M, Spaner DE, Galipeau J. Cytokine modulation of TLR expression and activation in mesenchymal stromal cells leads to a proinflammatory phenotype. J Immunol. 2009;182(12):7963–73.PubMedCrossRef Romieu-Mourez R, Francois M, Boivin MN, Bouchentouf M, Spaner DE, Galipeau J. Cytokine modulation of TLR expression and activation in mesenchymal stromal cells leads to a proinflammatory phenotype. J Immunol. 2009;182(12):7963–73.PubMedCrossRef
30.
go back to reference Das B, Kashino SS, Pulu I, Kalita D, Swami V, Yeger H, et al. CD271(+) bone marrow mesenchymal stem cells may provide a niche for dormant Mycobacterium tuberculosis. Sci Transl Med. 2013;5(170):170ra13.PubMedCrossRefPubMedCentral Das B, Kashino SS, Pulu I, Kalita D, Swami V, Yeger H, et al. CD271(+) bone marrow mesenchymal stem cells may provide a niche for dormant Mycobacterium tuberculosis. Sci Transl Med. 2013;5(170):170ra13.PubMedCrossRefPubMedCentral
31.
go back to reference Tornack J, Reece ST, Bauer WM, Vogelzang A, Bandermann S, Zedler U, et al. Human and mouse hematopoietic stem cells are a depot for dormant mycobacterium tuberculosis. PLoS One. 2017;12(1):e0169119.PubMedCrossRefPubMedCentral Tornack J, Reece ST, Bauer WM, Vogelzang A, Bandermann S, Zedler U, et al. Human and mouse hematopoietic stem cells are a depot for dormant mycobacterium tuberculosis. PLoS One. 2017;12(1):e0169119.PubMedCrossRefPubMedCentral
32.
go back to reference Mayito J, Andia I, Belay M, Jolliffe DA, Kateete DP, Reece ST, et al. Anatomic and cellular niches for mycobacterium tuberculosis in latent tuberculosis infection. J Infect Dis. 2019;219(5):685–94.PubMedCrossRef Mayito J, Andia I, Belay M, Jolliffe DA, Kateete DP, Reece ST, et al. Anatomic and cellular niches for mycobacterium tuberculosis in latent tuberculosis infection. J Infect Dis. 2019;219(5):685–94.PubMedCrossRef
33.
go back to reference Jain N, Kalam H, Singh L, Sharma V, Kedia S, Das P, et al. Mesenchymal stem cells offer a drug-tolerant and immune-privileged niche to mycobacterium tuberculosis. Nat Commun. 2020;11(1):3062.PubMedCrossRefPubMedCentral Jain N, Kalam H, Singh L, Sharma V, Kedia S, Das P, et al. Mesenchymal stem cells offer a drug-tolerant and immune-privileged niche to mycobacterium tuberculosis. Nat Commun. 2020;11(1):3062.PubMedCrossRefPubMedCentral
34.
go back to reference Fatima S, Kamble SS, Dwivedi VP, Bhattacharya D, Kumar S, Ranganathan A, et al. Mycobacterium tuberculosis programs mesenchymal stem cells to establish dormancy and persistence. J Clin Invest. 2020;130(2):655–61.PubMedCrossRef Fatima S, Kamble SS, Dwivedi VP, Bhattacharya D, Kumar S, Ranganathan A, et al. Mycobacterium tuberculosis programs mesenchymal stem cells to establish dormancy and persistence. J Clin Invest. 2020;130(2):655–61.PubMedCrossRef
35.
go back to reference Farrugia M, Baron B. The role of toll-like receptors in autoimmune diseases through failure of the self-recognition mechanism. Int J Inflam. 2017;2017:8391230.PubMedPubMedCentral Farrugia M, Baron B. The role of toll-like receptors in autoimmune diseases through failure of the self-recognition mechanism. Int J Inflam. 2017;2017:8391230.PubMedPubMedCentral
36.
37.
go back to reference Harding CV, Boom WH. Regulation of antigen presentation by mycobacterium tuberculosis: a role for toll-like receptors. Nat Rev Microbiol. 2010;8(4):296–307.PubMedCrossRefPubMedCentral Harding CV, Boom WH. Regulation of antigen presentation by mycobacterium tuberculosis: a role for toll-like receptors. Nat Rev Microbiol. 2010;8(4):296–307.PubMedCrossRefPubMedCentral
38.
go back to reference Mahomed H, Hawkridge T, Verver S, Geiter L, Hatherill M, Abrahams DA, et al. Predictive factors for latent tuberculosis infection among adolescents in a high-burden area in South Africa. Int J Tuberc Lung Dis. 2011;15(3):331–6.PubMed Mahomed H, Hawkridge T, Verver S, Geiter L, Hatherill M, Abrahams DA, et al. Predictive factors for latent tuberculosis infection among adolescents in a high-burden area in South Africa. Int J Tuberc Lung Dis. 2011;15(3):331–6.PubMed
39.
go back to reference Middelkoop K, Bekker LG, Liang H, Aquino LD, Sebastian E, Myer L, et al. Force of tuberculosis infection among adolescents in a high HIV and TB prevalence community: a cross-sectional observation study. BMC Infect Dis. 2011;11:156.PubMedCrossRefPubMedCentral Middelkoop K, Bekker LG, Liang H, Aquino LD, Sebastian E, Myer L, et al. Force of tuberculosis infection among adolescents in a high HIV and TB prevalence community: a cross-sectional observation study. BMC Infect Dis. 2011;11:156.PubMedCrossRefPubMedCentral
40.
go back to reference Ncayiyana JR, Bassett J, West N, Westreich D, Musenge E, Emch M, et al. Prevalence of latent tuberculosis infection and predictive factors in an urban informal settlement in Johannesburg, South Africa: a cross-sectional study. BMC Infect Dis. 2016;16(1):661.PubMedCrossRefPubMedCentral Ncayiyana JR, Bassett J, West N, Westreich D, Musenge E, Emch M, et al. Prevalence of latent tuberculosis infection and predictive factors in an urban informal settlement in Johannesburg, South Africa: a cross-sectional study. BMC Infect Dis. 2016;16(1):661.PubMedCrossRefPubMedCentral
42.
43.
go back to reference Jahantigh D, Salimi S, Alavi-Naini R, Emamdadi A, Owaysee Osquee H, Farajian MF. Association between TLR4 and TLR9 gene polymorphisms with development of pulmonary tuberculosis in Zahedan, southeastern Iran. Sci World J. 2013;2013:534053.CrossRef Jahantigh D, Salimi S, Alavi-Naini R, Emamdadi A, Owaysee Osquee H, Farajian MF. Association between TLR4 and TLR9 gene polymorphisms with development of pulmonary tuberculosis in Zahedan, southeastern Iran. Sci World J. 2013;2013:534053.CrossRef
44.
go back to reference Ma MJ, Xie LP, Wu SC, Tang F, Li H, Zhang ZS, et al. Toll-like receptors, tumor necrosis factor-alpha, and interleukin-10 gene polymorphisms in risk of pulmonary tuberculosis and disease severity. Hum Immunol. 2010;71(10):1005–10.PubMedCrossRef Ma MJ, Xie LP, Wu SC, Tang F, Li H, Zhang ZS, et al. Toll-like receptors, tumor necrosis factor-alpha, and interleukin-10 gene polymorphisms in risk of pulmonary tuberculosis and disease severity. Hum Immunol. 2010;71(10):1005–10.PubMedCrossRef
45.
go back to reference Krutzik SR, Modlin RL. The role of toll-like receptors in combating mycobacteria. Semin Immunol. 2004;16(1):35–41.PubMedCrossRef Krutzik SR, Modlin RL. The role of toll-like receptors in combating mycobacteria. Semin Immunol. 2004;16(1):35–41.PubMedCrossRef
Metadata
Title
Ex vivo mRNA expression of toll-like receptors during latent tuberculosis infection
Authors
Birhan Alemnew
Soren T. Hoff
Tamrat Abebe
Markos Abebe
Abraham Aseffa
Rawleigh Howe
Liya Wassie
Publication date
01-12-2021
Publisher
BioMed Central
Published in
BMC Immunology / Issue 1/2021
Electronic ISSN: 1471-2172
DOI
https://doi.org/10.1186/s12865-021-00400-4

Other articles of this Issue 1/2021

BMC Immunology 1/2021 Go to the issue