TY - JOUR
T1 - Detection of Mycobacterium tuberculosis complex DNA in CD34-positive peripheral blood mononuclear cells of asymptomatic tuberculosis contacts
T2 - an observational study
AU - Belay, Mulugeta
AU - Tulu, Begna
AU - Younis, Sidra
AU - Jolliffe, David A.
AU - Tayachew, Dawit
AU - Manwandu, Hana
AU - Abozen, Tenagnework
AU - Tirfie, Emawayish A.
AU - Tegegn, Metasebia
AU - Zewude, Aboma
AU - Forrest, Sally
AU - Mayito, Jonathan
AU - Huggett, Jim F.
AU - Jones, Gerwyn M.
AU - O'Sullivan, Denise M.
AU - Martineau, Henny M.
AU - Noursadeghi, Mahdad
AU - Chandran, Aneesh
AU - Harris, Kathryn A.
AU - Nikolayevskyy, Vlad
AU - Demaret, Julie
AU - Berg, Stefan
AU - Vordermeier, Martin
AU - Balcha, Taye T.
AU - Aseffa, Abraham
AU - Ameni, Gobena
AU - Abebe, Markos
AU - Reece, Stephen T.
AU - Martineau, Adrian R.
N1 - Funding Information:
This work was supported by a grant from the UK Medical Research Council to ARM (MR/P024548/1). SF and STR acknowledge support from the Cambridge National Institiute of Medical Research Biomedical Research Centre antibiotic resistance theme. JM acknowledges support from the Developing Excellence in Leadership, Training and Science Africa Initiative (DEL-15-011, through THRiVE-2). JFH, GMJ, and DMO'S acknowledge support from the UK government Department for Business, Energy & Industrial Strategy. MN acknowledges support from the Wellcome Trust (207511/Z/17/Z) and from National Institute for Health Research Biomedical Research Funding to University College London and University College London Hospital. We thank all the people who participated in the study, members of the field and laboratory teams who enrolled participants and processed samples, Susan Barton (Miltenyi Biotec) for assistance with flow cytometry, Eloise Busby (National Measurement Laboratory, LGC) for design of the femA_SE dPCR assay, and Pawan Dhami and Heli Vaikkinen for assistance with dPCR assays done at the Genomics and Genome Engineering Facility, University College London, which was supported by Cancer Research UK.
Funding Information:
This work was supported by a grant from the UK Medical Research Council to ARM (MR/P024548/1). SF and STR acknowledge support from the Cambridge National Institiute of Medical Research Biomedical Research Centre antibiotic resistance theme. JM acknowledges support from the Developing Excellence in Leadership, Training and Science Africa Initiative (DEL-15-011, through THRiVE-2). JFH, GMJ, and DMO'S acknowledge support from the UK government Department for Business, Energy & Industrial Strategy. MN acknowledges support from the Wellcome Trust (207511/Z/17/Z) and from National Institute for Health Research Biomedical Research Funding to University College London and University College London Hospital. We thank all the people who participated in the study, members of the field and laboratory teams who enrolled participants and processed samples, Susan Barton (Miltenyi Biotec) for assistance with flow cytometry, Eloise Busby (National Measurement Laboratory, LGC) for design of the femA_SE dPCR assay, and Pawan Dhami and Heli Vaikkinen for assistance with dPCR assays done at the Genomics and Genome Engineering Facility, University College London, which was supported by Cancer Research UK.
Publisher Copyright:
© 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license
PY - 2021/6
Y1 - 2021/6
N2 - Background: Haematopoietic stem cells expressing the CD34 surface marker have been posited as a niche for Mycobacterium tuberculosis complex bacilli during latent tuberculosis infection. Our aim was to determine whether M tuberculosis complex DNA is detectable in CD34-positive peripheral blood mononuclear cells (PBMCs) isolated from asymptomatic adults living in a setting with a high tuberculosis burden. Methods: We did a cross-sectional study in Ethiopia between Nov 22, 2017, and Jan 10, 2019. Digital PCR (dPCR) was used to determine whether M tuberculosis complex DNA was detectable in PBMCs isolated from 100 mL blood taken from asymptomatic adults with HIV infection or a history of recent household or occupational exposure to an index case of human or bovine tuberculosis. Participants were recruited from HIV clinics, tuberculosis clinics, and cattle farms in and around Addis Ababa. A nested prospective study was done in a subset of HIV-infected individuals to evaluate whether administration of isoniazid preventive therapy was effective in clearing M tuberculosis complex DNA from PBMCs. Follow-up was done between July 20, 2018, and Feb 13, 2019. QuantiFERON-TB Gold assays were also done on all baseline and follow-up samples. Findings: Valid dPCR data (ie, droplet counts >10 000 per well) were available for paired CD34-positive and CD34-negative PBMC fractions from 197 (70%) of 284 participants who contributed data to cross-sectional analyses. M tuberculosis complex DNA was detected in PBMCs of 156 of 197 participants with valid dPCR data (79%, 95% CI 74–85). It was more commonly present in CD34-positive than in CD34-negative fractions (154 [73%] of 197 vs 46 [23%] of 197; p<0·0001). Prevalence of dPCR-detected M tuberculosis complex DNA did not differ between QuantiFERON-negative and QuantiFERON-positive participants (77 [78%] of 99 vs 79 [81%] of 98; p=0·73), but it was higher in HIV-infected than in HIV-uninfected participants (67 [89%] of 75 vs 89 [73%] of 122, p=0·0065). By contrast, the proportion of QuantiFERON-positive participants was lower in HIV-infected than in HIV-uninfected participants (25 [33%] of 75 vs 73 [60%] of 122; p<0·0001). Administration of isoniazid preventive therapy reduced the prevalence of dPCR-detected M tuberculosis complex DNA from 41 (95%) of 43 HIV-infected individuals at baseline to 23 (53%) of 43 after treatment (p<0·0001), but it did not affect the prevalence of QuantiFERON positivity (17 [40%] of 43 at baseline vs 13 [30%] of 43 after treatment; p=0·13). Interpretation: We report a novel molecular microbiological biomarker of latent tuberculosis infection with properties that are distinct from those of a commercial interferon-γ release assay. Our findings implicate the bone marrow as a niche for M tuberculosis in latently infected individuals. Detection of M tuberculosis complex DNA in PBMCs has potential applications in the diagnosis of latent tuberculosis infection, in monitoring response to preventive therapy, and as an outcome measure in clinical trials of interventions to prevent or treat latent tuberculosis infection. Funding: UK Medical Research Council.
AB - Background: Haematopoietic stem cells expressing the CD34 surface marker have been posited as a niche for Mycobacterium tuberculosis complex bacilli during latent tuberculosis infection. Our aim was to determine whether M tuberculosis complex DNA is detectable in CD34-positive peripheral blood mononuclear cells (PBMCs) isolated from asymptomatic adults living in a setting with a high tuberculosis burden. Methods: We did a cross-sectional study in Ethiopia between Nov 22, 2017, and Jan 10, 2019. Digital PCR (dPCR) was used to determine whether M tuberculosis complex DNA was detectable in PBMCs isolated from 100 mL blood taken from asymptomatic adults with HIV infection or a history of recent household or occupational exposure to an index case of human or bovine tuberculosis. Participants were recruited from HIV clinics, tuberculosis clinics, and cattle farms in and around Addis Ababa. A nested prospective study was done in a subset of HIV-infected individuals to evaluate whether administration of isoniazid preventive therapy was effective in clearing M tuberculosis complex DNA from PBMCs. Follow-up was done between July 20, 2018, and Feb 13, 2019. QuantiFERON-TB Gold assays were also done on all baseline and follow-up samples. Findings: Valid dPCR data (ie, droplet counts >10 000 per well) were available for paired CD34-positive and CD34-negative PBMC fractions from 197 (70%) of 284 participants who contributed data to cross-sectional analyses. M tuberculosis complex DNA was detected in PBMCs of 156 of 197 participants with valid dPCR data (79%, 95% CI 74–85). It was more commonly present in CD34-positive than in CD34-negative fractions (154 [73%] of 197 vs 46 [23%] of 197; p<0·0001). Prevalence of dPCR-detected M tuberculosis complex DNA did not differ between QuantiFERON-negative and QuantiFERON-positive participants (77 [78%] of 99 vs 79 [81%] of 98; p=0·73), but it was higher in HIV-infected than in HIV-uninfected participants (67 [89%] of 75 vs 89 [73%] of 122, p=0·0065). By contrast, the proportion of QuantiFERON-positive participants was lower in HIV-infected than in HIV-uninfected participants (25 [33%] of 75 vs 73 [60%] of 122; p<0·0001). Administration of isoniazid preventive therapy reduced the prevalence of dPCR-detected M tuberculosis complex DNA from 41 (95%) of 43 HIV-infected individuals at baseline to 23 (53%) of 43 after treatment (p<0·0001), but it did not affect the prevalence of QuantiFERON positivity (17 [40%] of 43 at baseline vs 13 [30%] of 43 after treatment; p=0·13). Interpretation: We report a novel molecular microbiological biomarker of latent tuberculosis infection with properties that are distinct from those of a commercial interferon-γ release assay. Our findings implicate the bone marrow as a niche for M tuberculosis in latently infected individuals. Detection of M tuberculosis complex DNA in PBMCs has potential applications in the diagnosis of latent tuberculosis infection, in monitoring response to preventive therapy, and as an outcome measure in clinical trials of interventions to prevent or treat latent tuberculosis infection. Funding: UK Medical Research Council.
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U2 - 10.1016/S2666-5247(21)00043-4
DO - 10.1016/S2666-5247(21)00043-4
M3 - Article
AN - SCOPUS:85104932260
SN - 2666-5247
VL - 2
SP - e267-e275
JO - The Lancet Microbe
JF - The Lancet Microbe
IS - 6
ER -