Comparison of gene expression of immune mediators in lung and pulmonary lymph node granulomas from cattle experimentally infected with Mycobacterium bovis

The cellular infiltrates and macrophage activation pathways may differ in granulomas found in the lungs and pulmonary lymph nodes of cattle infected with Mycobacterium bovis. The aim of this study was to compare the histopathology and gene expression profiles of cytokines and immune mediators for cattle which had these lesions in both sites. Ten Friesian-cross, 15-16 month old cattle were challenged intratracheally with 5 × 10(3)CFU of virulent M. bovis and killed and necropsied at 28 weeks after infection. Seven animals were found to have gross TB granulomas in both their lungs and pulmonary lymph nodes (PLN) and these lesions were fully encapsulated with central necrosis and mineralisation. Neutrophil infiltration was clearly involved in granuloma in lung whereas neutrophils were limited in lesions of PLN. Comparisons were made of immune mediators from these two sites from the same animals as well as those between lesioned PLN tissues and non-lesioned prescapular lymph nodes (PSLN). Gene expressions of the immune mediators were normalised using a housekeeping gene (U1), a monocyte/macrophage marker (CD14) and a common leucocyte marker (CD45). mRNA expression of IFN-γ, IL-17A, IRF5(1) and arginase 1 (Arg1) was significantly up-regulated in lung compared to that for PLN (p<0.05), while mRNA expression of IFN-γ, IL-12p40, TNF-α and iNOs for PLN was significantly higher than that for PSLN (p<0.05). In addition, IL-10 mRNA expression was significantly higher for lung compared to PLN when normalised for CD45 (p<0.05). The results suggested that the stronger proinflammatory immune response in the lesioned lung may be a consequence of enhanced expression of IRF5 promoting IFN-γ and IL-17 production. In contrast, Arg1 expression in the lungs could facilitate the infection through competing with iNOs for l-arginine, preventing generation of nitric oxide for clearance of M. bovis infection.