Comparison of the roles of reactive oxygen and nitrogen intermediates in the host response to Mycobacterium tuberculosis using transgenic mice

Objective: To study the role of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) in host response to Mycobacterium tuberculosis. Design: M. tuberculosis infection (i.v.) was compared in B6 control and two strains of knockout (KO) mice. X-CGD mice with a nonfunctional allele for the gp91(phox) subunit of the phagocyte oxidase cytochrome b are unable to produce ROI whereas iNOS KO mice lack a functional inducible nitric oxide synthase (iNOS) gene and fail to make RNI. Results: M. tuberculosis growth was markedly enhanced in the lungs of X-CGD mice compared to B6 mice, but was controlled in the spleen and liver. In iNOS KO mice, M. tuberculosis growth was exacerbated in the spleen, but was unremarkable in the lungs compared to B6 mice until later (Day 60) in the infection. In vitro, X-CGD alveolar and peritoneal macrophages (MΦ) produced no ROI, but did produce RNI and inhibited growth of M. tuberculosis when activated with interferon gamma. iNOS KO MΦ produced ROI, but failed to produce RNI and could not cope with M. tuberculosis in vitro when activated. The inhibition of M. tuberculosis growth observed in activated B6 and X-CGD MΦ) was reversed in the presence of aminoguanidine. Conclusion: These KO mouse strains demonstrate the relative potent effects of ROI and RNI in resistance to M. tuberculosis and should prove useful for the study of regulatory and compensatory mechanisms of immunity.