Mycobacterial loads in lungs and spleen of Mtb-infected mice were determined at different time points post-infection as previously described (29 (link)). Lungs of Mtb-infected mice were fixed with 4% phosphate-buffered formalin, and 3 μm-thick sections were stained with either H&E or rabbit anti-mouse antibody specific for iNOS (Abcam) or rabbit anti-mouse IgA antibody (Abcam). Detection was performed using HRP-labelled anti-rabbit antibody (Dako) followed by 3, 3’-diaminobenzidine substrate (Dako). The lung images and lesion areas, iNOS and IgA positive areas were acquired in Nikon 90i Eclipse widefield microscope and quantified using NIS elements.
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