A whole blood assay as a simple, broad assessment of cytokines and chemokines to evaluate human immune responses to Mycobacterium tuberculosis antigens

dc.creatorSilva, Dolores
dc.creatorPonte, Carlos G. G.
dc.creatorHacker, Mariana A.
dc.creatorAntas, Paulo R. Z.
dc.date2015-07-03T12:34:37Z
dc.date2015-07-03T12:34:37Z
dc.date2013
dc.date.accessioned2023-09-26T21:15:22Z
dc.date.available2023-09-26T21:15:22Z
dc.identifierSILVA, Dolores et al. A whole blood assay as a simple, broad assessment of cytokines and chemokines to evaluate human immune responses to Mycobacterium tuberculosis antigens. Acta Tropica, v.127, n.2, p.85-81, aug. 2013.
dc.identifier0001-706X
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/11064
dc.identifier10.1016/j.actatropica.2013.04.002
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8871591
dc.descriptionIn vitro stimulation of whole blood or isolated peripheral blood cells with specific antigens is used for several purposes. We sought to identify a reliable, reproducible, fast and feasible in vitro method to assess human cellular immune responses to Mycobacterium tuberculosis. In contrast to peripheral blood mononuclear cell (PBMC) culture, a whole blood assay (WBA) provides a more physiological environment, which may provide a broader assessment of serum biomarker, biosignature profiles. Twenty-three asymptomatic individuals with M. tuberculosis infection were recruited. Total cells from the WBA (diluted 1:3 in completed RPMI) and PBMC (2 × 105 cells/ml) plus M. tuberculosis Ag85A, Ag85B, ESAT-6 and Mycobacterium bovis 65 kDa were characterized by flow cytometry, then added in 96-well plates and on day 5 plasma and supernatants were harvested for detection of 17 cytokines by a Luminex array system. There was agreement between PBMC and WBA for IL-2, IL-5, IL-6, IL-7, IL-13, IFN-γ, TNF-α, MCP-1 and MIP-1β. There was evidence toward higher IL-10 (p ≤ 0.049) and G-CSF (p ≤ 0.012) plasma production, and higher IL-1β (p ≤ 0.048), IL-4 (p ≤ 0.044), IL-12p70 (p ≤ 0.006), IL-17 (p ≤ 0.002) and GM-CSF (p ≤ 0.049) production for PBMC vs. WBA. Both methods provided virtually no reaction to the internal, negative control. Due to technical issues linked to data out of range, IL-8 data were not considered. These results suggest that, depending on the method employed, PBMC and/or WBA techniques provide fine conditions for the model proposed and thus whole blood cultures are well-suited low-cost proxy-measures during search for serum biomarkers.
dc.formatapplication/pdf
dc.languageeng
dc.publisherElsevier
dc.rightsrestricted access
dc.subjectMycobacterium tuberculosis
dc.subjectCytokines
dc.subjectChemokines
dc.subjectMycobacterium tuberculosis antigens
dc.subjectWhole blood assays
dc.subjectTuberculosis
dc.subjectCellularity
dc.subjectSerum biomarkers
dc.subjectTuberculose
dc.subjectBiomarcadores
dc.subjectAntígenos
dc.subjectQuimiocinas
dc.subjectCitocinas
dc.subjectMycobacterium tuberculosis
dc.titleA whole blood assay as a simple, broad assessment of cytokines and chemokines to evaluate human immune responses to Mycobacterium tuberculosis antigens
dc.typeArticle


Este ítem pertenece a la siguiente institución