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A novel mycobacteri...
A novel mycobacterial growth inhibition assay employing live-cell imaging of virulent M. tuberculosis and monitoring of host cell viability
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- Andersson, Blanka (författare)
- Linköpings universitet,Avdelningen för inflammation och infektion,Medicinska fakulteten
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- Jonsson Nordvall, Michaela (författare)
- Linköpings universitet,Avdelningen för inflammation och infektion,Medicinska fakulteten,Region Östergötland, Klinisk mikrobiologi
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- Welin, Amanda (författare)
- Linköpings universitet,Avdelningen för inflammation och infektion,Medicinska fakulteten
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- Lerm, Maria (författare)
- Linköpings universitet,Avdelningen för inflammation och infektion,Medicinska fakulteten
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- Schön, Thomas (författare)
- Linköpings universitet,Avdelningen för inflammation och infektion,Medicinska fakulteten,Department of Infectious Diseases and Clinical Microbiology, Kalmar County Hospital, Kalmar, Sweden
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(creator_code:org_t)
- CHURCHILL LIVINGSTONE, 2020
- 2020
- Engelska.
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Ingår i: Tuberculosis. - : CHURCHILL LIVINGSTONE. - 1472-9792 .- 1873-281X. ; 124
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Our aim was to develop a Mycobacterium tuberculosis (Mtb) growth inhibition assay (MGIA) as a summary estimate of host immune control of virulent Mtb. Mycobacterial growth inhibition (MGI) using previously frozen human PBMCs infected with H37Rv was assessed by live-cell imaging (Incucyte (c)) complemented by imaging flow cytometry analysis of phagocytosis. MGI measured as relative fluorescence units (RFU) was calibrated to time to positive culture (TTP) in BACTEC 960 MGIT. At a MOI (multiplicity of infection) of 5, there was a wide range of MGI of blood donors (1.1*10(6)-2.7*10(6) RFU, n = 14). Intra- and inter-assay variability were at most 17.5 and 20.7 CV%. Cell viability at day 5 was 57 and 62% monitored by the LDH and Draq7 assays respectively. There was a strong correlation between a readout for Mtb growth using CFU counts or TTP compared to RFU (r2 >= 0.96). Our MGIA enabling live-cell imaging and monitoring of cell viability was able to detect a wide range of Mtb growth inhibition by PBMCs and was calibrated to several readout options for bacterial growth. This MGIA may be valuable as a surrogate marker of host immunity in a personalized medicine approach.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Microbiology in the medical area (hsv//eng)
Nyckelord
- Tuberculosis; MGIA; Immune response; Live-cell imaging; PBMCs
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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