Mycobacterial antigen driven activation of CD14++ CD16-monocytes is a predictor of tuberculosis-associated immune reconstitution inflammatory syndrome

Paradoxical tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) is an aberrant inflammatory response occurring in a subset of TB-HIV co-infected patients initiating anti-retroviral therapy (ART). Here, we examined monocyte activation by prospectively quantitating pro-inflammatory plasma markers and monocyte subsets in TB-HIV co-infected patients from a South Indian cohort at baseline and following ART initiation at the time of IRIS, or at equivalent time points in non-IRIS controls. Pro-inflammatory biomarkers of innate and myeloid cell activation were increased in plasma of IRIS patients pre-ART and at the time of IRIS; this association was confirmed in a second cohort in South Africa. Increased expression of these markers correlated with elevated antigen load as measured by higher sputum culture grade and shorter duration of anti-TB therapy. Phenotypic analysis revealed the frequency of CD14++CD16− monocytes was an independent predictor of TB-IRIS, and was closely associated with plasma levels of CRP, TNF, IL-6 and tissue factor during IRIS. In addition, production of inflammatory cytokines by monocytes was higher in IRIS patients compared to controls pre-ART. These data point to a major role of mycobacterial antigen load and myeloid cell hyperactivation in the pathogenesis of TB-IRIS, and implicate monocytes and monocyte-derived cytokines as potential targets for TB-IRIS prevention or treatment

Inflammatory monocytes expressing tissue factor drive SIV and HIV coagulopathy

Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. http://www.sciencemag.org/about/science-licenses-journal-article-reuseThis is an article distributed under the terms of the Science Journals Default LicenseIn HIV infection, persistent inflammation despite effective antiretroviral therapy is linked to increased risk of noninfectious chronic complications such as cardiovascular and thromboembolic disease. A better understanding of inflammatory and coagulation pathways in HIV infection is needed to optimize clinical care. Markers of monocyte activation and coagulation independently predict morbidity and mortality associated with non-AIDS events. We identified a specific subset of monocytes that express tissue factor (TF), persist after virological suppression, and trigger the coagulation cascade by activating factor X. This subset of monocytes expressing TF had a distinct gene signature with up-regulated innate immune markers and evidence of robust production of multiple proinflammatory cytokines, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6, ex vivo and in vitro upon lipopolysaccharide stimulation. We validated our findings in a nonhuman primate model, showing that TF-expressing inflammatory monocytes were associated with simian immunodeficiency virus (SIV)-related coagulopathy in the progressive [pigtail macaques (PTMs)] but not in the nonpathogenic (African green monkeys) SIV infection model. Last, Ixolaris, an anticoagulant that inhibits the TF pathway, was tested and potently blocked functional TF activity in vitro in HIV and SIV infection without affecting monocyte responses to Toll-like receptor stimulation. Strikingly, in vivo treatment of SIV-infected PTMs with Ixolaris was associated with significant decreases in D-dimer and immune activation. These data suggest that TF-expressing monocytes are at the epicenter of inflammation and coagulation in chronic HIV and SIV infection and may represent a potential therapeutic target.This study was supported by the NIH Intramural Research Program, National Institute of Allergy and Infectious Diseases, and Bench-to-Bedside award R01HL117715-10S1 (to I.S. and I.P.). Part of this project has been also funded with federal funds from the National Cancer Institute, NIH, under contract no. HHSN261200800001E. The NHP study has also been funded in part with federal funds from the NIH (R01 HL123096 and RO1 HL117715 to I.P., R01 AI119346 to C.A., and R01AI104373 to R.M.R.).info:eu-repo/semantics/publishedVersio

Tissue factor expression by myeloid cells contributes to protective immune response against Mycobacterium tuberculosis infection: Innate Immunity

Tissue Factor (TF) is a transmembrane glycoprotein that plays an essential role in hemostasis by activating coagulation. TF is also expressed by monocytes/macrophages as part of the innate immune response to infections. In the current study, we determined the role of TF expressed by myeloid cells during Mycobacterium tuberculosis (M. tb) infection by using mice lacking the TF gene in myeloid cells (TFΔ) and human monocyte derived macrophages (MDMs). We found that during M. tb infection, a deficiency of TF in myeloid cells was associated with reduced inducible nitric oxide synthase (iNOS) expression, enhanced arginase 1 (Arg1) expression, enhanced IL-10 production and reduced apoptosis in infected macrophages, which augmented M. tb growth. Our results demonstrate that a deficiency of TF in myeloid cells promotes M2 like phenotype in M .tb infected macrophages. A deficiency in TF expression by myeloid cells was also associated with reduced fibrin deposition and increased matrix metalloproteases (MMP)-2 and MMP-9 mediated inflammation in M. tb infected lungs. Our studies demonstrate that TF expressed by myeloid cells has newly recognized abilities to polarize macrophages and to regulate M. tb growth

Tissue factor expression by myeloid cells contributes to protective immune response against Mycobacterium tuberculosis infection.

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2016-04-20T18:46:59Z No. of bitstreams: 1 Venkatasubramanian_Tissue factor....pdf: 2078232 bytes, checksum: 64ac10f6fa8886788693ba980ff0c7ee (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2016-04-20T19:03:54Z (GMT) No. of bitstreams: 1 Venkatasubramanian_Tissue factor....pdf: 2078232 bytes, checksum: 64ac10f6fa8886788693ba980ff0c7ee (MD5)Made available in DSpace on 2016-04-20T19:03:54Z (GMT). No. of bitstreams: 1 Venkatasubramanian_Tissue factor....pdf: 2078232 bytes, checksum: 64ac10f6fa8886788693ba980ff0c7ee (MD5) Previous issue date: 2016University of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Cellular and Molecular Biology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Cellular and Molecular Biology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USAFrederick National Laboratory for Cancer Research. Clinical Research Directorate/Clinical Monitoring Research Program. Leidos Biomedical Research, Inc. Frederick, MD, USAFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório Integrado de Microbiologia e Imunoregulação. Salvador, BA, Brasil / Brazilian Institute for Tuberculosis Research. Research Center. Salvador, BA, BrasilThe University of North Carolina at Chapel Hill School of Medicine. Department of Medicine. NC, USAUniversity of Texas Health Science Center at Tyler. Department of Cellular and Molecular Biology. Tyler, TX, USAUniversity of Texas Health Science Center at Tyler. Department of Pulmonary Immunology. Tyler, TX, USATissue factor (TF) is a transmembrane glycoprotein that plays an essential role in hemostasis by activating coagulation. TF is also expressed by monocytes/macrophages as part of the innate immune response to infections. In the current study, we determined the role of TF expressed by myeloid cells during Mycobacterium tuberculosis (M. tb) infection by using mice lacking the TF gene in myeloid cells (TF(Δ) ) and human monocyte derived macrophages (MDMs). We found that during M. tb infection, a deficiency of TF in myeloid cells was associated with reduced inducible nitric oxide synthase (iNOS) expression, enhanced arginase 1 (Arg1) expression, enhanced IL-10 production and reduced apoptosis in infected macrophages, which augmented M. tb growth. Our results demonstrate that a deficiency of TF in myeloid cells promotes M2-like phenotype in M .tb infected macrophages. A deficiency in TF expression by myeloid cells was also associated with reduced fibrin deposition and increased matrix metalloproteases (MMP)-2 and MMP-9 mediated inflammation in M. tb infected lungs. Our studies demonstrate that TF expressed by myeloid cells has newly recognized abilities to polarize macrophages and to regulate M. tb growt

Mycobacterial Antigen Driven Activation of CD14++CD162 Monocytes Is a Predictor of Tuberculosis-Associated Immune Reconstitution Inflammatory Syndrome

Submitted by Nuzia Santos ([email protected]) on 2015-03-17T17:08:53Z No. of bitstreams: 1 2014_157.pdf: 8039547 bytes, checksum: 61886898f8eac99f8b159f84a0aed9a8 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-17T17:09:01Z (GMT) No. of bitstreams: 1 2014_157.pdf: 8039547 bytes, checksum: 61886898f8eac99f8b159f84a0aed9a8 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-17T17:17:24Z (GMT) No. of bitstreams: 1 2014_157.pdf: 8039547 bytes, checksum: 61886898f8eac99f8b159f84a0aed9a8 (MD5)Made available in DSpace on 2015-03-17T17:17:24Z (GMT). No. of bitstreams: 1 2014_157.pdf: 8039547 bytes, checksum: 61886898f8eac99f8b159f84a0aed9a8 (MD5) Previous issue date: 2014National Institutes of Health. National Institute of Allergy and Infectious Diseases. Laboratory of Parasitic Diseases. Immunobiology Section. Bethesda, MD, United States of AmericaNational Institutes of Health. National Institute of Allergy and Infectious Diseases.Laboratory of Immunoregulation. Clinical and Molecular Retrovirology Section. Bethesda, MD, United States of AmericaNational Institute for Research in Tuberculosis. Chennai, IndiaNational Institutes of Health. National Institute of Allergy and Infectious Diseases.Laboratory of Immunoregulation. Clinical and Molecular Retrovirology Section. Bethesda, MD, United States of AmericaNational Institute for Research in Tuberculosis. Chennai, IndiaNational Institute for Research in Tuberculosis. Chennai, IndiaNational Institute for Research in Tuberculosis. Chennai, IndiaNational Institutes of Health. National Institute of Allergy and Infectious Diseases.Laboratory of Immunoregulation. Clinical and Molecular Retrovirology Section. Bethesda, MD, United States of AmericaNational Institutes of Health. National Institute of Allergy and Infectious Diseases.Laboratory of Immunoregulation. Clinical and Molecular Retrovirology Section. Bethesda, MD, United States of AmericaLaboratório de Imunopatologia, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, Minas Gerais, BrazilUniversity of Cape Town. Institute of Infectious Disease and Molecular Medicine, and Department of Medicine. Clinical Infectious Diseases Research Initiative. Cape Town, South AfricaUniversity of Cape Town. Institute of Infectious Disease and Molecular Medicine, and Department of Medicine. Clinical Infectious Diseases Research Initiative. Cape Town, South Africa/Imperial College London. Department of Medicine. London, United Kingdom/MRC National Institute for Medical Research. London, United KingdomUniversity of Cape Town. Institute of Infectious Disease and Molecular Medicine, and Department of Medicine. Clinical Infectious Diseases Research Initiative. Cape Town, South Africa/Imperial College London. Department of Medicine. London, United KingdomUniversity of Cape Town. Institute of Infectious Disease and Molecular Medicine, and Department of Medicine. Clinical Infectious Diseases Research Initiative. Cape Town, South AfricaNational Institutes of Health. National Institute of Allergy and Infectious Diseases. Biostatistics Research Branch. Bethesda, MD, United States of AmericaNational Institutes of Health. National Institute of Allergy and Infectious Diseases. T-Lymphocyte Biology Unit, Laboratory of Parasitic Diseases. Bethesda, MD, United States of AmericaNational Institute for Research in Tuberculosis. Chennai, IndiaNational Institutes of Health. National Institute of Allergy and Infectious Diseases. Laboratory of Parasitic Diseases. Immunobiology Section. Bethesda, MD, United States of AmericaNational Institutes of Health. National Institute of Allergy and Infectious Diseases.Laboratory of Immunoregulation. Clinical and Molecular Retrovirology Section. Bethesda, MD, United States of AmericaParadoxical tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) is an aberrant inflammatory response occurring in a subset of TB-HIV co-infected patients initiating anti-retroviral therapy (ART). Here, we examined monocyte activation by prospectively quantitating pro-inflammatory plasma markers and monocyte subsets in TB-HIV co-infected patients from a South Indian cohort at baseline and following ART initiation at the time of IRIS, or at equivalent time points in non-IRIS controls. Pro-inflammatory biomarkers of innate and myeloid cell activation were increased in plasma of IRIS patients pre-ART and at the time of IRIS; this association was confirmed in a second cohort in South Africa. Increased expression of these markers correlated with elevated antigen load as measured by higher sputum culture grade and shorter duration of anti-TB therapy. Phenotypic analysis revealed the frequency of CD14++CD16− monocytes was an independent predictor of TB-IRIS, and was closely associated with plasma levels of CRP, TNF, IL-6 and tissue factor during IRIS. In addition, production of inflammatory cytokines by monocytes was higher in IRIS patients compared to controls pre-ART. These data point to a major role of mycobacterial antigen load and myeloid cell hyperactivation in the pathogenesis of TB-IRIS, and implicate monocytes and monocyte-derived cytokines as potential targets for TB-IRIS prevention or treatment

Chronic fluoxetine treatment selectively uncouples raphe 5-HT(1A) receptors as measured by [(35)S]-GTPγS autoradiography

1. Selective Serotonin Reuptake Inhibitors (SSRIs) are thought to have a delay in therapeutic efficacy because of the need to overcome the inhibitory influence of raphe 5-HT(1A) autoreceptors. Prolonged SSRI administration has been reported to desensitize these autoreceptors. We have used [(35)S]-GTPγS autoradiography to determine whether this desensitization occurs at the level of receptor/G protein coupling. 2. Male mice were injected intraperitoneally once a day with saline or 20 mg kg(−1) fluoxetine for either 2 days or 14 days. 5-HT(1A) receptor binding and coupling to G proteins were assessed using [(3)H]-8-OH-DPAT and [(35)S]-GTPγS autoradiography, respectively. 3. The 5-HT receptor agonist 5-carboxamidotryptamine (5-CT) stimulated [(35)S]-GTPγS binding in the substantia nigra, as well as in hippocampus and dorsal raphe nucleus. The 5-HT(1A) receptor antagonist p-MPPF (4-fluoro-N-(2-[4-(2-methoxyphenyl)1-piperazinyl]ethyl)-N-(2-pyridinyl)benzamide) blocked this effect in the latter regions, whereas the 5-HT(1B/D) antagonist GR-127,935 (2′-methyl-4′-(5-methyl-[1,2,4]oxadiazol-3-yl)-biphenyl-4-carboxylic acid [4-methoxy-3-(4-methyl-piperazin-l-yl)-phenyl]-amide) only decreased labelling in substantia nigra. 4. Fourteen-day fluoxetine treatment decreased 5-CT-stimulated [(35)S]-GTPγS binding in dorsal raphe (saline: 112±12% stimulation; fluoxetine: 66±13%), but not in substantia nigra (99±14% vs 103±7%) or hippocampus (157±3% vs 148±18%). Two-day fluoxetine treatment did not alter 5-CT-stimulated [(35)S]-GTPγS binding in any of the brain areas investigated. 5. Decreased [(35)S]-GTPγS binding was not due to receptor down-regulation, since the density of raphe [(3)H]-8-OH-DPAT binding sites was unaffected by fluoxetine treatment. 6. These results suggest that the desensitization of presynaptic 5-HT(1A) receptor function occurs at the level of receptor-G protein interaction on dorsal raphe neurons, and may underlie the therapeutic efficacy of long-term SSRI treatment

Circulating monocyte counts do not correlate with soluble biomarkers of inflammation and immune activation in TB-HIV co-infected individuals.

<p>(<b>A</b>) Numbers of circulating monocytes (left panel) and neutrophils (right panel) are compared at week 0 (pre-ART), at weeks 6 or at the time of IRIS, and at week 24 after ART initiation between TB-HIV co-infected patients who developed paradoxical TB-IRIS (n = 26) and those who did not (n = 22). Lines represent median values and interquartile ranges. Data were analyzed using the Mann-Whitney test or Wilcoxon matched-pairs test for paired comparisons within each study group. ** P<0.01, *** P<0.001. (<b>B</b>) The network analysis (interactome) showed statistically significant correlations (P<0.05) between neutrophil or monocyte counts and plasma biomarkers. Associations were assessed with Spearman rank tests.</p