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An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis.
O'Leary, Seónadh; Bahlool, Ahmad Z; O'Connor, Gemma; Cryan, Sally-Ann; Keane, Joseph M; O'Sullivan, Mary P.
  • O'Leary S; Department of Clinical Medicine, Trinity Translational Medicine Institute, St. James's Hospital, Trinity College Dublin, The University of Dublin.
  • Bahlool AZ; Department of Clinical Medicine, Trinity Translational Medicine Institute, St. James's Hospital, Trinity College Dublin, The University of Dublin; School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland (RCSI).
  • O'Connor G; School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland (RCSI).
  • Cryan SA; School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland (RCSI); SFI Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD; SFI Centre for Research in Medical Devices (CURAM), RCSI, Dublin and National University of Ireland.
  • Keane JM; Department of Clinical Medicine, Trinity Translational Medicine Institute, St. James's Hospital, Trinity College Dublin, The University of Dublin.
  • O'Sullivan MP; Department of Clinical Medicine, Trinity Translational Medicine Institute, St. James's Hospital, Trinity College Dublin, The University of Dublin; mary.osullivan@tcd.ie.
J Vis Exp ; (174)2021 08 16.
Article in English | MEDLINE | ID: covidwho-1378456
ABSTRACT
Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), was the most significant infectious disease killer globally until the advent of COVID-19. Mtb has evolved to persist in its intracellular environment, evade host defenses, and has developed resistance to many anti-tubercular drugs. One approach to solving resistance is identifying existing approved drugs that will boost the host immune response to Mtb. These drugs could then be repurposed as adjunctive host-directed therapies (HDT) to shorten treatment time and help overcome antibiotic resistance. Quantification of intracellular Mtb growth in macrophages is a crucial aspect of assessing potential HDT. The gold standard for measuring Mtb growth is counting colony-forming units (CFU) on agar plates. This is a slow, labor-intensive assay that does not lend itself to rapid screening of drugs. In this protocol, an automated, broth-based culture system, which is more commonly used to detect Mtb in clinical specimens, has been adapted for preclinical screening of host-directed therapies. The capacity of the liquid culture assay system to investigate intracellular Mtb growth in macrophages treated with HDT was evaluated. The HDTs tested for their ability to inhibit Mtb growth were all-trans Retinoic acid (AtRA), both in solution and encapsulated in poly(lactic-co-glycolic acid) (PLGA) microparticles and the combination of interferon-gamma and linezolid. The advantages of this automated liquid culture-based technique over the CFU method include simplicity of setup, less labor-intensive preparation, and faster time to results (5-12 days compared to 21 days or more for agar plates).
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Tuberculosis / Mycobacterium tuberculosis Type of study: Diagnostic study / Experimental Studies / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Tuberculosis / Mycobacterium tuberculosis Type of study: Diagnostic study / Experimental Studies / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article