Evaluation of Human Antimycobacterial Immunity Using Recombinant Reporter Mycobacteria

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Abstract

A novel in vitro whole blood model was developed to study human antimycobacterial immunity. Recombinant reporter mycobacteria were used to enumerate the bacteria, and interactions between host immune cells and mycobacteria were studied using whole blood rather than cell fractions. The ability of healthy tuberculin-positive and tuberculin-negative individuals to restrict mycobacterial growth was compared. Growth of luminescent mycobacteria was significantly lower in blood samples of tuberculin-positive individuals than in blood samples of tuberculin-negative individuals (P = .005). Restricted mycobacterial growth was associated with significantly higher production of tumor necrosis factor (TNF)—α and interferon (IFN)—γ (P = .01 and .004, respectively). Inhibition of the TNF-α and IFN-γ response pathways by neutralizing monoclonal antibodies increased mycobacterial growth in whole blood. This model is the first functional assay in which individual variations in cellmediated immunity are shown to correlate with differences in ability to control mycobacterial growth. It provides a new tool for studying human mycobactericidal mechanisms and, potentially, for the evaluation of improved vaccines.

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