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dc.contributor.authorMekonnen, Meseret Habtamuen_US
dc.contributor.authorAbebe, Markosen_US
dc.contributor.authorAseffa, Abrahamen_US
dc.contributor.authorDyrhol-Riise, Anne Maen_US
dc.contributor.authorSpurkland, Anneen_US
dc.contributor.authorAbrahamsen, Gregeren_US
dc.PublishedMekonnen MH, Abebe M, Aseffa A, Dyrhol-Riise AM, Spurkland A, Abrahamsen G. In vitro analysis of antigen induced T cell-monocyte conjugates by imaging flow cytometry. JIM - Journal of Immunological Methods. 2018;460:93-100eng
dc.descriptionUnder embargo until: 04.07.2019en_US
dc.description.abstractThere is a lack of suitable correlates of immune protection against Mycobacterium tuberculosis (Mtb) infection. T cells and monocytes play key roles in host immunity against Mtb. Thus, a method that allows assessing their interaction would contribute to the understanding of immune regulation in tuberculosis (TB). We have established imaging flow cytometer (IFC) based in vitro assay for the analysis of early events in T cell-monocyte interaction, upstream of cytokine production and T cell proliferation. This was achieved through short term stimulation of peripheral blood mononuclear cells (PBMC) from healthy Norwegian blood donors with Mycobacterium bovis Bacille Calmette-Guérin (BCG). In our assay, we examined the kinetics of BCG uptake by monocytes using fluorescently labeled BCG and T cell-monocyte interaction based on synapse formation (CD3/TCR polarization). Our results showed that BCG stimulation induced a gradual increase in the proportion of conjugated T cells displaying NF-κB translocation to the nucleus in a time dependent manner, with the highest frequency observed at 6 h. We subsequently tested PBMC from a small cohort of active TB patients (n = 7) and observed a similar BCG induced NF-κB translocation in T cells conjugated with monocytes. The method allowed for simultaneous evaluation of T cell-monocyte conjugates and T cell activation as measured by NF-κB translocation, following short-term challenge of human PBMC with BCG. Whether this novel approach could serve as a diagnostic or prognostic marker needs to be investigated using a wide array of Mtb specific antigens in a larger cohort of patients with different TB infection status.en_US
dc.rightsAttribution CC BY-NC-NDeng
dc.subjectImaging flow cytometeryeng
dc.subjectNuclear factor κappa-Beng
dc.titleIn vitro analysis of antigen induced T cell-monocyte conjugates by imaging flow cytometryen_US
dc.typePeer reviewed
dc.typeJournal article
dc.rights.holderCopyright 2018 Elsevier
dc.source.journalJIM - Journal of Immunological Methods

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