According to preliminary results of the immunocytochemical and the immunofluorescence assay, the production rates for fibronectin, type I collagen, filamentous actin, and vimentin in granuloma macrophages were not affected by mycobacterial loads, nor were those in granuloma fibroblasts from mice following infection with BCG vaccinein vivoor in peritoneal macrophages and splenic fibroblasts from your control group of intact mice (Figures 12(a)C12(e) and ?and13).13). [2C4]. In 2014, 480,000 new cases of with multiple drug resistance were diagnosed, of which only 48% recovered . At present, there is the only anti-TB vaccine called the Bacillus Calmette-Gurin (BCG) prepared from an attenuated live strain ofM. bovisM. tuberculosisby aerosol transmission. Pulmonary macrophages entrap mycobacteria by phagocytosis and eliminate them in phagolysosomes using active forms of oxygen and nitrogen, lysosomal hydrolases, and harmful peptides in a low-pH medium. The proinflammatory cytokines IFNM. tuberculosisin chronic granulomatous inflammatory lesions largely composed of macrophages [2, 5, 6]. Low BCG-mycobacterial loads in animal organs and tissues at different time points of chronic contamination experienced previously been established by bacteriological methods in a model of latent tuberculous contamination under which mice were infected with BCG-mycobacteriain vivo[7C10]. Using our initial model of mouse granulomas inex vivoculture, we have, for the first time, decided the bacterial weight in macrophages, dendritic cells, and multinucleate Langhans giant cells in individual granulomas obtained from mice with latent tuberculous contamination afterin vivoexposure to BCG vaccine [11, 12]. In some host cells, not only did BCG-mycobacteria survive, but also they were actively reproducing and created cording colonies, cording being the Zapalog indication of their virulence . Interestingly, there was a difference in behavior between mycobacteria of virulent and nonvirulent strains inin vitrocultures of infected human, mouse, and cow cells [13C18]. Mycobacteria of virulent strains were actively reproducing in cells infectedin vitroM. tuberculosisof nonvirulent strains were basically found in vacuoles before they were damaged there within 2C7 days of observationin vitro. However, there are very few comparative studies of associations between mycobacteria of different strains and host cells in animals infectedin vivoor following acute infectionin vitro[19, 20]. And very few are the studies researching associations between BCG-mycobacteria and host cells [11, 12, 19, 21]. As is known, BCG vaccines Rabbit polyclonal to IL29 can occasionally cause severe disease in children with inborn errors of immunity often referred to as BCG-osis [22, 23]. Importantly, clinical observations of BCG contamination (including BCG adenitis) in AIDS patients after as many as 30 years following BCG vaccination are still being discussed . Therefore, understanding associations between BCG-mycobacteria and host cells both after infectionin vivoand after acute infectionin vitrois important for studying the development of BCG-induced anti-TB immunity, developing better BCG-based vaccines [5, 6], and screening vaccine candidates in animal models , including mouse models Zapalog of tuberculous and nontuberculous mycobacterial infections [24, 25]. In the present work, we conducted a comparative study of the mycobacterial loads in granuloma cells from your bone marrow and spleens of mice with latent tuberculous contamination following contamination with BCGin vivoand several days ofex vivoculture and in the cultures of bone marrow cells and peritoneal macrophages obtained from intact mice and infected with BCGin vitroin vitroand the death of cells having increased BCG loads. Throughout 48C120?h ofex vivoculture, mouse granuloma macrophages each basically remained to contain a single BCG organism, and increased numbers of such microorganisms in Zapalog some macrophages did not cause the host cells to die. Analysis of the levels of the proinflammatory cytokines IFNand IL-1ex lover vivoandin vitrocultures suggested that even though active production of these molecules in mouse granuloma cells did not help in Zapalog eliminating all mycobacteria in the host cells, it helped in restricting mycobacterial reproduction in granuloma macrophages. By contrast, a considerable increase in the Zapalog number of BCG-mycobacteria was observed in thosein vitroinfected mouse bone marrow and peritoneal macrophages, whether alive or lifeless by apoptosis/necrosis, in which no active synthesis of these markers was going on. 2. Materials and Methods 2.1. Animals Two-month-old BALB/c male mice were obtained from the Animal Breeding Facility of the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences (Novosibirsk, Russia). Mice were bred and managed under standard vivarium conditions, with water and food providedad libitumM. bovis(the Bacillus Calmette-Gurin vaccine, BCG-1, the Institute of Microbiology and Epidemiology, Moscow, Russia) at a dose of 0.5?mg per mouse, which amounted to 3 106 viable BCG-mycobacteria in 0.9% NaCl solution. Twenty-four mice were each infected via tail.