Characterization of Immune Responses of Human PBMCs Infected with Mycobacterium Tuberculosis H37Ra: Impact of Donor Declared BCG Vaccination History on Immune Responses and M. Tuberculosis Growth

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2018 Sep 12

PMID 30204787

Citations 6

Authors

Sudha Bhavanam

Gina R Rayat

Monika Keelan

Dennis Kunimoto

Steven J Drews

Affiliations

Soon will be listed here.

Abstract

This study characterized the immune responses in early Mycobacterium tuberculosis (Mtb) H37Ra infection of human peripheral blood mononuclear cell (PBMC)-collagen matrix culture and the impact of Bacille Calmette-Guérin (BCG) vaccination history of donor PBMCs on the immune responses to Mtb infection. Aggregates of PBMCs were initially observed on day 3 and the size of aggregates continued to increase on day 8 post-infection, where macrophages and T cell subsets were identified to be present. Similarly, mycobacterial load progressively increased in infected PBMCs during the 8 days of culture but were significantly lower in infected PBMCs from BCG vaccinated (BCG+) donors compared to unvaccinated (BCG-) donors. The levels of INF-γ, TNF-α, IL-4, IL-6, IL-10 and IL-17 in the supernatants of Mtb-infected PBMCs peaked at day 3 and decreased on days 5 and 8. The levels of these cytokines except IL-10 were significantly lower in Mtb-infected PBMCs from BCG+ donors compared to infected PBMCs from BCG- donors. The percentages of activated naïve Th cells, activated effector memory Th cells and activated central memory Tc cells were significantly higher in Mtb-infected PBMCs compared to uninfected PBMCs at day 8 post-infection. Further, the proportion of activated central memory Tc cells was significantly higher in infected PBMCs from BCG+ donors compared to the BCG- donors. This study highlights the possibility that BCG vaccination may confound results that utilize human PBMCs to study Mtb infection.

Citing Articles

Effect of BCG Revaccination on Occupationally Exposed Medical Personnel Vaccinated against SARS-CoV-2.

Ramos-Martinez E, Falfan-Valencia R, Perez-Rubio G, Andrade W, Rojas-Serrano J, Ambrocio-Ortiz E Cells. 2021; 10(11).

PMID: 34831403 PMC: 8617982. DOI: 10.3390/cells10113179.

In Vitro Miniaturized Tuberculosis Spheroid Model.

Mukundan S, Singh P, Shah A, Kumar R, ONeill K, Carter C Biomedicines. 2021; 9(9).

PMID: 34572395 PMC: 8470281. DOI: 10.3390/biomedicines9091209.

Modelling early events in Mycobacterium bovis infection using a co-culture model of the bovine alveolus.

Lee D, Stewart G, Chambers M Sci Rep. 2020; 10(1):18495.

PMID: 33116165 PMC: 7595104. DOI: 10.1038/s41598-020-75113-6.

Coordinated In Vitro Release of Granulysin, Perforin and IFN-γ in TB and HIV/TB Co-Infection Associated with Clinical Outcomes before and after Anti-TB Treatment.

Pitabut N, Dhepakson P, Sakurada S, Keicho N, Khusmith S Pathogens. 2020; 9(8).

PMID: 32823923 PMC: 7459825. DOI: 10.3390/pathogens9080655.

Profiles of Local and Systemic Inflammation in the Outcome of Treatment of Human Cutaneous Leishmaniasis Caused by ().

Navas A, Fernandez O, Gallego-Marin C, Castro M, Rosales-Chilama M, Murillo J Infect Immun. 2019; 88(3).

PMID: 31818959 PMC: 7035935. DOI: 10.1128/IAI.00764-19.

References

Cowan J, Pandey S, Filion L, Angel J, Kumar A, Cameron D . Comparison of interferon-γ-, interleukin (IL)-17- and IL-22-expressing CD4 T cells, IL-22-expressing granulocytes and proinflammatory cytokines during latent and active tuberculosis infection. Clin Exp Immunol. 2012; 167(2):317-29. PMC: 3278699. DOI: 10.1111/j.1365-2249.2011.04520.x. View

Bhavanam S, Rayat G, Keelan M, Kunimoto D, Drews S . Understanding the pathophysiology of the human TB lung granuloma using in vitro granuloma models. Future Microbiol. 2016; 11:1073-89. DOI: 10.2217/fmb-2016-0005. View

McGeachy M, Bak-Jensen K, Chen Y, Tato C, Blumenschein W, Mcclanahan T . TGF-beta and IL-6 drive the production of IL-17 and IL-10 by T cells and restrain T(H)-17 cell-mediated pathology. Nat Immunol. 2007; 8(12):1390-7. DOI: 10.1038/ni1539. View

Freeman S, Post F, Bekker L, Harbacheuski R, Steyn L, Ryffel B . Mycobacterium tuberculosis H37Ra and H37Rv differential growth and cytokine/chemokine induction in murine macrophages in vitro. J Interferon Cytokine Res. 2006; 26(1):27-33. DOI: 10.1089/jir.2006.26.27. View

Lockhart E, Green A, Flynn J . IL-17 production is dominated by gammadelta T cells rather than CD4 T cells during Mycobacterium tuberculosis infection. J Immunol. 2006; 177(7):4662-9. DOI: 10.4049/jimmunol.177.7.4662. View

Al-Attiyah R, Madi N, El-Shamy A, Wiker H, Andersen P, Mustafa A . Cytokine profiles in tuberculosis patients and healthy subjects in response to complex and single antigens of Mycobacterium tuberculosis. FEMS Immunol Med Microbiol. 2006; 47(2):254-61. DOI: 10.1111/j.1574-695X.2006.00110.x. View

Maggioli M, Palmer M, Thacker T, Vordermeier H, McGill J, Whelan A . Increased TNF-α/IFN-γ/IL-2 and Decreased TNF-α/IFN-γ Production by Central Memory T Cells Are Associated with Protective Responses against Bovine Tuberculosis Following BCG Vaccination. Front Immunol. 2016; 7:421. PMC: 5066095. DOI: 10.3389/fimmu.2016.00421. View

Giacomini E, Remoli M, Scandurra M, Gafa V, Pardini M, Fattorini L . Expression of proinflammatory and regulatory cytokines via NF-κB and MAPK-dependent and IFN regulatory factor-3-independent mechanisms in human primary monocytes infected by Mycobacterium tuberculosis. Clin Dev Immunol. 2011; 2011:841346. PMC: 3010679. DOI: 10.1155/2011/841346. View

Flynn J, Gideon H, Mattila J, Lin P . Immunology studies in non-human primate models of tuberculosis. Immunol Rev. 2015; 264(1):60-73. PMC: 4339213. DOI: 10.1111/imr.12258. View

10.

Dardalhon V, Awasthi A, Kwon H, Galileos G, Gao W, Sobel R . IL-4 inhibits TGF-beta-induced Foxp3+ T cells and, together with TGF-beta, generates IL-9+ IL-10+ Foxp3(-) effector T cells. Nat Immunol. 2008; 9(12):1347-55. PMC: 2999006. DOI: 10.1038/ni.1677. View

11.

Li L, Qiao D, Zhang X, Liu Z, Wu C . The immune responses of central and effector memory BCG-specific CD4+ T cells in BCG-vaccinated PPD+ donors were modulated by Treg cells. Immunobiology. 2010; 216(4):477-84. DOI: 10.1016/j.imbio.2010.09.003. View

12.

Bonecini-Almeida M, Ho J, Boechat N, Huard R, Chitale S, Doo H . Down-modulation of lung immune responses by interleukin-10 and transforming growth factor beta (TGF-beta) and analysis of TGF-beta receptors I and II in active tuberculosis. Infect Immun. 2004; 72(5):2628-34. PMC: 387880. DOI: 10.1128/IAI.72.5.2628-2634.2004. View

13.

Chavez-Galan L, Ocana-Guzman R, Torre-Bouscoulet L, Garcia-de-Alba C, Sada-Ovalle I . Exposure of Monocytes to Lipoarabinomannan Promotes Their Differentiation into Functionally and Phenotypically Immature Macrophages. J Immunol Res. 2015; 2015:984973. PMC: 4548148. DOI: 10.1155/2015/984973. View

14.

Puissegur M, Botanch C, Duteyrat J, Delsol G, Caratero C, Altare F . An in vitro dual model of mycobacterial granulomas to investigate the molecular interactions between mycobacteria and human host cells. Cell Microbiol. 2004; 6(5):423-33. DOI: 10.1111/j.1462-5822.2004.00371.x. View

15.

Kheir A, Alhaj A, Ibrahim S . The sensitivity of BCG scar as an indicator of previous vaccination among Sudanese infants. Vaccine. 2011; 29(46):8189-91. DOI: 10.1016/j.vaccine.2011.08.095. View

16.

Braian C, Svensson M, Brighenti S, Lerm M, Parasa V . A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection. J Vis Exp. 2015; (104). PMC: 4692636. DOI: 10.3791/53084. View

17.

Ernst J . Macrophage receptors for Mycobacterium tuberculosis. Infect Immun. 1998; 66(4):1277-81. PMC: 108049. DOI: 10.1128/IAI.66.4.1277-1281.1998. View

18.

Sakaguchi S . Naturally arising CD4+ regulatory t cells for immunologic self-tolerance and negative control of immune responses. Annu Rev Immunol. 2004; 22:531-62. DOI: 10.1146/annurev.immunol.21.120601.141122. View

19.

Zwerling A, Behr M, Verma A, Brewer T, Menzies D, Pai M . The BCG World Atlas: a database of global BCG vaccination policies and practices. PLoS Med. 2011; 8(3):e1001012. PMC: 3062527. DOI: 10.1371/journal.pmed.1001012. View

20.

Sinha P, Gupta A, Prakash P, Anupurba S, Tripathi R, Srivastava G . Differentiation of Mycobacterium tuberculosis complex from non-tubercular mycobacteria by nested multiplex PCR targeting IS6110, MTP40 and 32kD alpha antigen encoding gene fragments. BMC Infect Dis. 2016; 16:123. PMC: 4788904. DOI: 10.1186/s12879-016-1450-1. View