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Microbiology 150 (2004), 1507-1518; DOI  10.1099/mic.0.26815-0
© 2004 Society for General Microbiology

Induction of Mycobacterium avium growth restriction and inhibition of phagosome–endosome interactions during macrophage activation and apoptosis induction by picolinic acid plus IFN{gamma}

Teresa F. Pais{dagger} and Rui Appelberg{ddagger}

Laboratory of Microbiology and Immunology of Infection, Institute for Molecular and Cell Biology, Rua do Campo Alegre 823, 4150-180 Porto, Portugal

Correspondence
Rui Appelberg
rappelb{at}ibmc.up.pt

Treatment of mouse macrophages with picolinic acid (PA) and {gamma}-interferon (IFN{gamma}) led to the restriction of Mycobacterium avium proliferation concomitant with the sequential acquisition of metabolic changes typical of apoptosis, mitochondrial depolarization, annexin V staining and caspase activation, over a period of up to 5 days. However, triggering of cell death by ATP, staurosporine or H2O2 failed to affect mycobacterial viability. In contrast to untreated macrophages where extensive interactions between phagosomes and endosomes were observed, phagosomes from treated macrophages lost the ability to acquire endosomal dextran. N-Acetylcysteine was able to revert both the anti-mycobacterial activity of treated macrophages as well as the block in phagosome–endosome interactions. The treatment, however, induced only a minor increase in the acquisition of lysosomal markers, namely Lamp-1, and did not increase to any great extent the acidification of the phagosomes. These data thus suggest that the anti-mycobacterial activity of PA and IFN{gamma} depends on the interruption of intracellular vesicular trafficking, namely the blocking of acquisition of endosomal material by the microbe.

Abbreviations: FACS, fluorescence-activated cell sorting; Jc-1, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide; IFN{gamma}, {gamma}-interferon; MTT, methylthiazoletetrazolium; PA, picolinic acid; PS, phosphatidylserine; TMR, tetramethylrhodamine

{dagger}Present address: Instituto Gulbenkian de Ciência, Oeiras, Portugal.

{ddagger}Present address: ICBAS, Instituto de Ciências Biome'dicas Abel Salazar, Porto, Portugal.




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