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(a) Schematic overview of the study design. Peripheral blood mononuclear cells (PBMC) from healthy controls (HC) or patients diagnosed with infectious clinical TB (TB) were either exposed to Mycobacterium tuberculosis ( Mtb )-derived stimuli prior flow cytometry analysis (b) or infected with Mtb <t>H37Rv,</t> prior embedding within an extra-cellular matrix for 3D ex-vivo granuloma formation and assessment of the indicated microbiological read-outs (c-d). (b) Dot plot displaying background subtracted frequency of IFNγ-producing CD4 T cells measured by flow cytometry after overnight stimulation with ESAT-6/CFP- 10/TB7.7 peptide pool, Mtb soluble-cell wall proteins (SCWP) or Mtb whole-cell lysate (WCL); lines indicate median. (c) Bacterial load in colony forming units (CFU), eight days post-infection (p.i.). (d) Frequency of dormant-like Mtb based on auramine-O/Nile red dual staining, one day p.i.; bars depict the median. All p values represent Mann-Whitney tests.
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Simian immunodeficiency virus (SIV) infection impairs humoral immunity. Change from baseline (prior to any infection) in plasma humoral profiles (IgG, IgG1, IgG2, IgG3, FcγRIIa and FcγRIIIa) 8 weeks following infection with Mycobacterium tuberculosis ( Mtb ) in MCM without SIV (naïve; n = 8; yellow left bar) or coinfected with SIV (coinfection; n = 7; red right bar) as determined by multiplexing to Mtb <t>soluble</t> <t>cell</t> <t>wall,</t> cell wall fraction, cell membrane, cytosol fraction, TX114 <t>proteins,</t> peptidoglycan and cell lysate. Change in median fluorescence intensity data was normalised via z ‐scoring before being represented as a heatmap (red indicates higher levels and blue indicates lower levels). Multiplex assays were repeated in duplicate.
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Simian immunodeficiency virus (SIV) infection impairs humoral immunity. Change from baseline (prior to any infection) in plasma humoral profiles (IgG, IgG1, IgG2, IgG3, FcγRIIa and FcγRIIIa) 8 weeks following infection with Mycobacterium tuberculosis ( Mtb ) in MCM without SIV (naïve; n = 8; yellow left bar) or coinfected with SIV (coinfection; n = 7; red right bar) as determined by multiplexing to Mtb <t>soluble</t> <t>cell</t> <t>wall,</t> cell wall fraction, cell membrane, cytosol fraction, TX114 <t>proteins,</t> peptidoglycan and cell lysate. Change in median fluorescence intensity data was normalised via z ‐scoring before being represented as a heatmap (red indicates higher levels and blue indicates lower levels). Multiplex assays were repeated in duplicate.
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Simian immunodeficiency virus (SIV) infection impairs humoral immunity. Change from baseline (prior to any infection) in plasma humoral profiles (IgG, IgG1, IgG2, IgG3, FcγRIIa and FcγRIIIa) 8 weeks following infection with Mycobacterium tuberculosis ( Mtb ) in MCM without SIV (naïve; n = 8; yellow left bar) or coinfected with SIV (coinfection; n = 7; red right bar) as determined by multiplexing to Mtb <t>soluble</t> <t>cell</t> <t>wall,</t> cell wall fraction, cell membrane, cytosol fraction, TX114 <t>proteins,</t> peptidoglycan and cell lysate. Change in median fluorescence intensity data was normalised via z ‐scoring before being represented as a heatmap (red indicates higher levels and blue indicates lower levels). Multiplex assays were repeated in duplicate.
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(a) Schematic overview of the study design. Peripheral blood mononuclear cells (PBMC) from healthy controls (HC) or patients diagnosed with infectious clinical TB (TB) were either exposed to Mycobacterium tuberculosis ( Mtb )-derived stimuli prior flow cytometry analysis (b) or infected with Mtb H37Rv, prior embedding within an extra-cellular matrix for 3D ex-vivo granuloma formation and assessment of the indicated microbiological read-outs (c-d). (b) Dot plot displaying background subtracted frequency of IFNγ-producing CD4 T cells measured by flow cytometry after overnight stimulation with ESAT-6/CFP- 10/TB7.7 peptide pool, Mtb soluble-cell wall proteins (SCWP) or Mtb whole-cell lysate (WCL); lines indicate median. (c) Bacterial load in colony forming units (CFU), eight days post-infection (p.i.). (d) Frequency of dormant-like Mtb based on auramine-O/Nile red dual staining, one day p.i.; bars depict the median. All p values represent Mann-Whitney tests.

Journal: bioRxiv

Article Title: Granulysin antimicrobial activity promotes dormancy in Mycobacterium tuberculosis

doi: 10.1101/2024.09.27.615427

Figure Lengend Snippet: (a) Schematic overview of the study design. Peripheral blood mononuclear cells (PBMC) from healthy controls (HC) or patients diagnosed with infectious clinical TB (TB) were either exposed to Mycobacterium tuberculosis ( Mtb )-derived stimuli prior flow cytometry analysis (b) or infected with Mtb H37Rv, prior embedding within an extra-cellular matrix for 3D ex-vivo granuloma formation and assessment of the indicated microbiological read-outs (c-d). (b) Dot plot displaying background subtracted frequency of IFNγ-producing CD4 T cells measured by flow cytometry after overnight stimulation with ESAT-6/CFP- 10/TB7.7 peptide pool, Mtb soluble-cell wall proteins (SCWP) or Mtb whole-cell lysate (WCL); lines indicate median. (c) Bacterial load in colony forming units (CFU), eight days post-infection (p.i.). (d) Frequency of dormant-like Mtb based on auramine-O/Nile red dual staining, one day p.i.; bars depict the median. All p values represent Mann-Whitney tests.

Article Snippet: Mtb H37Rv whole cell lysate (NR-14822) and Mtb H37Rv soluble cell wall proteins (NR- 14840) were obtained through BEI Resources, NIAID, NIH.

Techniques: Derivative Assay, Flow Cytometry, Infection, Ex Vivo, Staining, MANN-WHITNEY

Simian immunodeficiency virus (SIV) infection impairs humoral immunity. Change from baseline (prior to any infection) in plasma humoral profiles (IgG, IgG1, IgG2, IgG3, FcγRIIa and FcγRIIIa) 8 weeks following infection with Mycobacterium tuberculosis ( Mtb ) in MCM without SIV (naïve; n = 8; yellow left bar) or coinfected with SIV (coinfection; n = 7; red right bar) as determined by multiplexing to Mtb soluble cell wall, cell wall fraction, cell membrane, cytosol fraction, TX114 proteins, peptidoglycan and cell lysate. Change in median fluorescence intensity data was normalised via z ‐scoring before being represented as a heatmap (red indicates higher levels and blue indicates lower levels). Multiplex assays were repeated in duplicate.

Journal: Clinical & Translational Immunology

Article Title: Antibody glycosylation correlates with disease progression in SIV‐ Mycobacterium tuberculosis coinfected cynomolgus macaques

doi: 10.1002/cti2.1474

Figure Lengend Snippet: Simian immunodeficiency virus (SIV) infection impairs humoral immunity. Change from baseline (prior to any infection) in plasma humoral profiles (IgG, IgG1, IgG2, IgG3, FcγRIIa and FcγRIIIa) 8 weeks following infection with Mycobacterium tuberculosis ( Mtb ) in MCM without SIV (naïve; n = 8; yellow left bar) or coinfected with SIV (coinfection; n = 7; red right bar) as determined by multiplexing to Mtb soluble cell wall, cell wall fraction, cell membrane, cytosol fraction, TX114 proteins, peptidoglycan and cell lysate. Change in median fluorescence intensity data was normalised via z ‐scoring before being represented as a heatmap (red indicates higher levels and blue indicates lower levels). Multiplex assays were repeated in duplicate.

Article Snippet: Mtb antigens of the H37Rv strain (sourced from BEI resources) are as follows: soluble cell wall proteins (NR‐14840), cell wall fraction (NR‐14828), cell membrane proteins (NR‐14831), cytosol fraction (NR‐14834), purified peptidoglycan (NR‐14853) and TX‐114 soluble proteins (NR‐14831).

Techniques: Virus, Infection, Clinical Proteomics, Multiplexing, Membrane, Fluorescence, Multiplex Assay