cathepsin-g and Granuloma

The hallmark of human Mycobacterium tuberculosis infection is the presence of lung granulomas. Lung granulomas can have different phenotypes, with caseous necrosis and hypoxia present within these structures during active tuberculosis. Production of NO by the inducible host enzyme NOS2 is a key antimycobacterial defense mechanism that requires oxygen as a substrate; it is therefore likely to perform inefficiently in hypoxic regions of granulomas in which M. tuberculosis persists. Here we have used Nos2-/- mice to investigate host-protective mechanisms within hypoxic granulomas and identified a role for host serine proteases in hypoxic granulomas in determining outcome of disease. Nos2-/- mice reproduced human-like granulomas in the lung when infected with M. tuberculosis in the ear dermis. The granulomas were hypoxic and contained large amounts of the serine protease cathepsin G and clade B serine protease inhibitors (serpins). Extrinsic inhibition of serine protease activity in vivo resulted in distorted granuloma structure, extensive hypoxia, and increased bacterial growth in this model. These data suggest that serine protease activity acts as a protective mechanism within hypoxic regions of lung granulomas and present a potential new strategy for the treatment of tuberculosis.

Topics: Animals; Cathepsin G; Granuloma; Hypoxia; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Mycobacterium tuberculosis; Necrosis; Pulmonary Fibrosis; Serine Proteases; Tuberculosis, Pulmonary

Proteoglycan (GAG) and collagen are lost from cartilage juxtaposed to murine granulomatous tissue in both control and C57bg/bg (elastase deficient mice). The objective was to extract and characterise proteoglycan degrading activity within granulomas of both strains.. 15 animals (female C57b1/6 and C57bg/bg mice) per group were used.. Cotton-wrapped rat femoral head cartilages were implanted subcutaneously into the dorsum of the mice and the granulomas excised fourteen days later.. Granuloma and granuloma cell-granule preparations were fractionated within a detergent-based buffer and tested for their abilities to degrade cartilage in vitro in the presence and absence of enzyme inhibitors. Elastase and cathepsin G activities were also assessed using specific substrates. Statistical significance was calculated using Student's t-test.. Extracts from both strains induced the loss of cartilage GAG. This was correlated with cathepsin G activity (r = 0.96) and was inhibited by a specific cathepsin-G inhibitor (95%, p < 0.001), but not specific elastase or metalloproteinase inhibitors. Elastase activity but not that of cathepsin G was absent in the beige mice, whilst both enzymes were active in the controls.. It appears that neutrophil cathepsin G may play an important role in the degradation of cartilage proteoglycan in the murine cotton-pellet granuloma in both C57b1/6 and C57bg/bg.

Topics: Animals; Cartilage; Cathepsin G; Cathepsins; Female; Granuloma; Male; Mice; Mice, Inbred C57BL; Pancreatic Elastase; Proteoglycans; Rats; Rats, Wistar; Serine Endopeptidases