Expression and cellular source of 11-βHSD1, 11-βHSD2 and glucocorticoids in the lungs and liver during late active pulmonary TB
Groups of five mice with T2D, TB and T2D/TB were euthanized by exsanguination under pentobarbital anesthesia after 60 and 120 days of infection, in two independent experiments. The late active disease is well established by this time in this model, as characterized by extensive pneumonia, high bacillary loads and low expression of IFN-γ and TNF-α. Three right lungs were intra-tracheally perfused with absolute ethanol and after 24hr the tissue was embedded for histology and immunohistochemistry studies, while the left lungs were immediately frozen for total RNA isolation and gene expression determination of 11-βHSD1 and 11-βHSD2 by RT-PCR. The same tissue samples for the histology study were used to determine by immunohistochemistry the cellular source of corticosterone and cortisone and their concentrations by digital pathology. Tissue fragments from the liver were processed in the same way.