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  1. Article ; Online: Mycobacterial-specific secretion of cytokines and chemokines in healthcare workers with apparent resistance to infection with

    Shey, Muki Shehu / Balfour, Avuyonke / Masina, Nomawethu / Bekiswa, Abulele / Schutz, Charlotte / Goliath, Rene / Dielle, Rachel / Katoto, Patrick Dmc / Wilkinson, Katalin Andrea / Lewinsohn, David / Lewinsohn, Deborah Anne / Meintjes, Graeme

    Frontiers in immunology

    2023  Volume 14, Page(s) 1176615

    Abstract: Background: Currently, diagnosis of latent TB infection (LTBI) is based on the secretion of IFN-γ in response to : Methods: We enrolled HIV-uninfected healthcare workers who had worked in high TB-exposure environments for 5 years or longer. We ... ...

    Abstract Background: Currently, diagnosis of latent TB infection (LTBI) is based on the secretion of IFN-γ in response to
    Methods: We enrolled HIV-uninfected healthcare workers who had worked in high TB-exposure environments for 5 years or longer. We screened them for LTBI using the tuberculin skin test and the QuantiFERON-TB Gold Plus assay. We performed multiplex Luminex to measure concentrations of T cell-associated cytokines and chemokines as well as total antibodies in plasma collected from unstimulated fresh whole blood and supernatants from QuantiFERON-TB Gold Plus tubes following incubation of whole blood for 16-24 hours with ESAT6/CFP10 peptides.
    Results: Samples from 78 individuals were analyzed: 33 resisters (TST<10mm; IGRA<0.35 IU/mL), 33 with LTBI (TST≥10mm and IGRA≥0.35 IU/mL) and 12 discordant (TST=0mm; IGRA≥1.0 IU/mL). There were no differences in concentrations of cytokines and chemokines in plasma between the different groups. Resisters had significantly lower concentrations of IFN-γ, IL-2, TNF-α, MIP-1α, MIP-1β, ITAC, IL-13 and GM-CSF in supernatants compared with LTBI group. There were no significant differences in the concentrations in supernatants of IL-10, IL-1β, IL-17A, IL-21, IL-23, MIP-3α, IL-4, IL-5, IL-6, IL-7, IL-8, Fractalkine and IL-12p70 between the groups. We observed that resisters had similar concentrations of total antibodies (IgG1, IgG2, IgG3, IgG4, IgA, and IgM) in plasma and supernatants compared to the LTBI and discordant groups.
    Conclusion: Resistance to Mtb infection despite sustained exposure is associated with lower Mtb-specific secretion of Th1-associated cytokines and chemokines. However, resisters showed secreted concentrations after Mtb stimulation of total antibodies and cytokines/chemokines associated with innate and Th17 immune responses similar to those with Mtb infection. This suggests an ability to mount non-IFN-γ immune responses to Mtb in apparent resisters.
    MeSH term(s) Humans ; Mycobacterium tuberculosis ; Cytokines ; Tuberculosis ; Latent Tuberculosis ; Tuberculin Test ; Latent Infection
    Chemical Substances Cytokines
    Language English
    Publishing date 2023-05-18
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2606827-8
    ISSN 1664-3224 ; 1664-3224
    ISSN (online) 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2023.1176615
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Contribution of APCs to mucosal-associated invariant T cell activation in infectious disease and cancer.

    Shey, Muki Shehu / Balfour, Avuyonke / Wilkinson, Katalin Andrea / Meintjes, Graeme

    Innate immunity

    2018  Volume 24, Issue 4, Page(s) 192–202

    Abstract: APCs such as monocytes and dendritic cells are among the first cells to recognize invading pathogens and initiate an immune response. The innate response can either eliminate the pathogen directly, or through presentation of Ags to T cells, which can ... ...

    Abstract APCs such as monocytes and dendritic cells are among the first cells to recognize invading pathogens and initiate an immune response. The innate response can either eliminate the pathogen directly, or through presentation of Ags to T cells, which can help to clear the infection. Mucosal-associated invariant T (MAIT) cells are among the unconventional T cells whose activation does not involve the classical co-stimulation during Ag presentation. MAIT cells can be activated either via presentation of unconventional Ags (such as riboflavin metabolites) through the evolutionarily conserved major histocompatibility class I-like molecule, MR1, or directly by cytokines such as IL-12 and IL-18. Given that APCs produce cytokines and can express MR1, these cells can play an important role in both pathways of MAIT cell activation. In this review, we summarize evidence on the role of APCs in MAIT cell activation in infectious disease and cancer. A better understanding of the interactions between APCs and MAIT cells is important in further elucidating the role of MAIT cells in infectious diseases, which may facilitate the design of novel interventions such as vaccines.
    MeSH term(s) Antigen Presentation ; Antigen-Presenting Cells/immunology ; Antigen-Presenting Cells/metabolism ; Communicable Diseases/immunology ; Histocompatibility Antigens Class I/metabolism ; Humans ; Interleukin-12/metabolism ; Interleukin-18/metabolism ; Lymphocyte Activation ; Mucosal-Associated Invariant T Cells/immunology ; Mucosal-Associated Invariant T Cells/metabolism ; Neoplasms/immunology ; Receptors, Antigen, T-Cell, alpha-beta/metabolism
    Chemical Substances Histocompatibility Antigens Class I ; Interleukin-18 ; Receptors, Antigen, T-Cell, alpha-beta ; Interleukin-12 (187348-17-0)
    Language English
    Publishing date 2018-04-09
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ISSN 1753-4267
    ISSN (online) 1753-4267
    DOI 10.1177/1753425918768695
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Xpert Ultra testing of blood in severe HIV-associated tuberculosis to detect and measure Mycobacterium tuberculosis blood stream infection: a diagnostic and disease biomarker cohort study.

    Boloko, Linda / Schutz, Charlotte / Sibiya, Nomfundo / Balfour, Avuyonke / Ward, Amy / Shey, Muki / Nicol, Mark P / Burton, Rosie / Wilkinson, Robert J / Maartens, Gary / Meintjes, Graeme / Barr, David A

    The Lancet. Microbe

    2022  Volume 3, Issue 7, Page(s) e521–e532

    Abstract: Background: Mycobacterium tuberculosis bloodstream infection is a leading cause of death in people living with HIV and disseminated bacillary load might be a key driver of disease severity. We aimed to assess Xpert MTB/RIF Ultra (Xpert Ultra) testing of ...

    Abstract Background: Mycobacterium tuberculosis bloodstream infection is a leading cause of death in people living with HIV and disseminated bacillary load might be a key driver of disease severity. We aimed to assess Xpert MTB/RIF Ultra (Xpert Ultra) testing of blood as a diagnostic for M tuberculosis bloodstream infection and investigate cycle threshold as a quantitative disease biomarker.
    Methods: In this cohort study, we obtained biobanked blood samples from a large and well characterised cohort of adult patients admitted to hospital in Western Cape, South Africa with suspected HIV-associated tuberculosis and a CD4 count less than 350 cells per μL. Patients already receiving antituberculosis therapy were excluded. Samples were obtained on recruitment within 72 h of admission to hospital, and patients were followed up for 12 weeks to determine survival. We tested the biobanked blood samples using the Xpert Ultra platform after lysis and wash processing of the blood. We assessed diagnostic yield (proportion of cases detected, with unavailable test results coded as negative) against a microbiological reference, both as a function of markers of critical-illness and compared with other rapid diagnostics (urine lipoarabinomannan and sputum Xpert). Quantitative blood Xpert Ultra results were evaluated as a disease biomarker by assessing association with disease phenotype defined by principal component analysis of 32 host-response markers. Prognostic value compared to other tuberculosis biomarkers was assessed using likelihood ratio testing of nested models predicting 12-week mortality.
    Findings: Between Jan 16, 2014, and Oct 19, 2016, of the 659 participants recruited to the parent study, 582 had an available biobanked blood sample. 447 (77%) of 582 met the microbiological reference standard for tuberculosis diagnosis. Median CD4 count was 62 (IQR 221-33) cells per μL, and 123 (21%) of participants died by 12-weeks follow-up. Blood Xpert Ultra was positive in 165 (37%) of 447 participants with confirmed tuberculosis by the microbiological reference standard, with a diagnostic yield of 0·37 (95% CI 0·32-0·42). Diagnostic yield increased with lower CD4 count or haemoglobin, and outperformed urine lipoarabinomannan testing in participants with elevated venous lactate. Quantitative blood Xpert Ultra results were more closely associated with mortality than other tuberculosis biomarkers including blood culture, and urine lipoarabinomannan, or urine Xpert (all p<0·05). A principal component of clinical phenotype capturing markers of inflammation, tissue damage, and organ dysfunction was strongly associated with both blood Xpert-Ultra positivity (associated with a SD increase of 1·1 in PC score, p<0·0001) and cycle threshold (r= -0·5; p<0·0001).
    Interpretation: Xpert Ultra testing of pre-processed blood could be used as a rapid diagnostic test in critically ill patients with suspected HIV-associated tuberculosis, while also giving additional prognostic information compared with other available markers. A dose-response relationship between quantitative blood Xpert Ultra results, host-response phenotype, and mortality risk adds to evidence that suggests M tuberculosis bloodstream infection bacillary load is causally related to outcomes.
    Funding: Wellcome Trust, National Institute of Health Fogarty International Center, South African MRC, UK National Institute of Health Research, National Research Foundation of South Africa.
    Translations: For the Xhosa and Afrikaans translations of the abstract see Supplementary Materials section.
    MeSH term(s) Bacteremia/complications ; Biomarkers ; Cohort Studies ; HIV Infections/complications ; Humans ; Mycobacterium tuberculosis/genetics ; Sensitivity and Specificity ; Sepsis/complications ; Tuberculosis/diagnosis ; Tuberculosis, Pulmonary/diagnosis
    Chemical Substances Biomarkers
    Language English
    Publishing date 2022-05-26
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ISSN 2666-5247
    ISSN (online) 2666-5247
    DOI 10.1016/S2666-5247(22)00062-3
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Rifampicin resistance and mortality in patients hospitalised with HIV-associated tuberculosis.

    Spies, Ruan / Schutz, Charlotte / Ward, Amy / Balfour, Avuyonke / Shey, Muki / Nicol, Mark / Burton, Rosie / Sossen, Bianca / Wilkinson, Robert / Barr, David / Meintjes, Graeme

    Southern African journal of HIV medicine

    2022  Volume 23, Issue 1, Page(s) 1396

    Abstract: Background: Patients with HIV and drug-resistant tuberculosis (TB) are at high risk of death.: Objectives: We investigated the association between rifampicin-resistant TB (RR-TB) and mortality in a cohort of patients who were admitted to hospital at ... ...

    Abstract Background: Patients with HIV and drug-resistant tuberculosis (TB) are at high risk of death.
    Objectives: We investigated the association between rifampicin-resistant TB (RR-TB) and mortality in a cohort of patients who were admitted to hospital at the time of TB diagnosis.
    Method: Adults hospitalised at Khayelitsha Hospital and diagnosed with HIV-associated TB during admission, were enrolled between 2013 and 2016. Clinical, biochemical and microbiological data were prospectively collected and participants were followed up for 12 weeks.
    Results: Participants with microbiologically confirmed TB (
    Conclusion: Mortality at 12 weeks in participants with RR-TB was high compared to participants with RS-TB. Delays in the initiation of appropriate treatment and poorer regimen efficacy are proposed as contributors to higher mortality in hospitalised patients with HIV and RR-TB.
    Language English
    Publishing date 2022-09-27
    Publishing country South Africa
    Document type Journal Article
    ZDB-ID 2259791-8
    ISSN 2078-6751 ; 2078-6751
    ISSN (online) 2078-6751
    ISSN 2078-6751
    DOI 10.4102/sajhivmed.v23i1.1396
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Functional and Activation Profiles of Mucosal-Associated Invariant T Cells in Patients With Tuberculosis and HIV in a High Endemic Setting.

    Balfour, Avuyonke / Schutz, Charlotte / Goliath, Rene / Wilkinson, Katalin A / Sayed, Sumaya / Sossen, Bianca / Kanyik, Jean-Paul / Ward, Amy / Ndzhukule, Rhandzu / Gela, Anele / Lewinsohn, David M / Lewinsohn, Deborah A / Meintjes, Graeme / Shey, Muki

    Frontiers in immunology

    2021  Volume 12, Page(s) 648216

    Abstract: Background: ...

    Abstract Background:
    MeSH term(s) AIDS-Related Opportunistic Infections/epidemiology ; AIDS-Related Opportunistic Infections/immunology ; AIDS-Related Opportunistic Infections/virology ; Adult ; Antitubercular Agents/therapeutic use ; Case-Control Studies ; Cross-Sectional Studies ; Endemic Diseases ; Female ; Flow Cytometry ; HIV-1/isolation & purification ; HLA-DR Antigens/metabolism ; Humans ; Immunity, Mucosal ; Interferon-gamma/metabolism ; Latent Infection/epidemiology ; Latent Infection/immunology ; Latent Infection/microbiology ; Lymphocyte Activation/drug effects ; Lymphocyte Activation/immunology ; Lymphocyte Count ; Male ; Mucosal-Associated Invariant T Cells/drug effects ; Mucosal-Associated Invariant T Cells/immunology ; Mycobacterium tuberculosis/genetics ; Mycobacterium tuberculosis/isolation & purification ; South Africa/epidemiology ; Treatment Outcome ; Tuberculosis, Pulmonary/drug therapy ; Tuberculosis, Pulmonary/epidemiology ; Tuberculosis, Pulmonary/immunology ; Tuberculosis, Pulmonary/microbiology ; Young Adult
    Chemical Substances Antitubercular Agents ; HLA-DR Antigens ; IFNG protein, human ; Interferon-gamma (82115-62-6)
    Language English
    Publishing date 2021-03-22
    Publishing country Switzerland
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2606827-8
    ISSN 1664-3224 ; 1664-3224
    ISSN (online) 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2021.648216
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Serial measurement of M. tuberculosis in blood from critically-ill patients with HIV-associated tuberculosis.

    Barr, David A / Schutz, Charlotte / Balfour, Avuyonke / Shey, Muki / Kamariza, Mireille / Bertozzi, Carolyn R / de Wet, Timothy J / Dinkele, Ryan / Ward, Amy / Haigh, Kathryn A / Kanyik, Jean-Paul / Mizrahi, Valerie / Nicol, Mark P / Wilkinson, Robert J / Lalloo, David G / Warner, Digby F / Meintjes, Graeme / Davies, Gerry

    EBioMedicine

    2022  Volume 78, Page(s) 103949

    Abstract: Background: Despite being highly prevalent in hospitalised patients with severe HIV-associated tuberculosis (TB) and sepsis, little is known about the mycobacteriology of Mycobacterium tuberculosis bloodstream infection (MTBBSI). We developed methods to ...

    Abstract Background: Despite being highly prevalent in hospitalised patients with severe HIV-associated tuberculosis (TB) and sepsis, little is known about the mycobacteriology of Mycobacterium tuberculosis bloodstream infection (MTBBSI). We developed methods to serially measure bacillary load in blood and used these to characterise MTBBSI response to anti-TB therapy (ATT) and relationship with mortality.
    Methods: We established a microscopy method for direct visualisation of M. tuberculosis bacilli in blood using a novel lysis-concentration protocol and the fluorescent probe, 4-N,N-dimethylaminonaphthalimide-trehalose (DMN-Tre). We tested blood using GeneXpert® MTB/RIF-Ultra (Xpert-ultra) and Myco/F lytic culture after processing blood through lysis-wash steps to remove PCR inhibitors and anti-microbial drug carry-over. HIV-positive patients predicted to have MTBBSI gave blood samples 0, 4, 24, 48 and 72 h after ATT initiation. Bacillary loads were quantified using microscopy, Xpert-ultra cycle threshold, and culture time-to-positivity. Pharmacodynamics were modelled using these measures combined on an ordinal scale, including association with 12-week mortality.
    Findings: M. tuberculosis was detected in 27 of 28 recruited participants; 25 (89%) by blood Xpert-ultra, 22 (79%) by DMN-Tre microscopy, and 21 (75%) by Myco/F lytic blood culture. Eight (29%) participants died by 12-week follow-up. In a combined pharmacodynamic model, predicted probabilities of negative DMN-Tre microscopy, blood Xpert-ultra, or blood culture after 72 h treatment were 0·64, 0·27, and 0·94, respectively, in those who survived, compared with 0·23, 0·06, and 0·71 in those who died (posterior probability of slower clearance of MTBBSI in those that died >0·99). DMN-Tre microscopy of blood demonstrated heterogenous bacillary morphologies, including microcolonies and clumps. Bacillary cell-length varied significantly with ATT exposure (mean cell-length increase 0·13 log-µm/day; 95%CrI 0·10-0·16).
    Interpretation: Pharmacodynamics of MTBBSI treatment can be captured using DMN-Tre microscopy, blood Xpert-ultra and culture. This could facilitate interventional trials in severe HIV-associated TB.
    Funding: Wellcome Trust, NIH Fogarty International Center, South African MRC, NIHR(UK), National Research Foundation of South Africa.
    MeSH term(s) Critical Illness ; HIV Infections/complications ; HIV Infections/drug therapy ; Humans ; Mycobacterium tuberculosis ; Sensitivity and Specificity ; Sputum/microbiology ; Tuberculosis/complications ; Tuberculosis/diagnosis ; Tuberculosis/drug therapy ; Tuberculosis, Pulmonary/microbiology
    Language English
    Publishing date 2022-03-21
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2851331-9
    ISSN 2352-3964
    ISSN (online) 2352-3964
    DOI 10.1016/j.ebiom.2022.103949
    Database MEDical Literature Analysis and Retrieval System OnLINE

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