Figure 1.
Animals, immunization and challenge schedule.
(a) BALB/c mice were immunized intraperitoneally (i.p.) with BCG or rBCG-LTAK63 (1 x 106 CFU). At 24 h and 7 d, peritoneal cells were recovered for the evaluation of cell recruitment, production of Nitric Oxide (NO), Hydrogen peroxide (H2O2) and inflammatory cytokines (without in vitro stimulation). (b) BALB/c mice were immunized subcutaneously (s.c.) with BCG or rBCG-LTAK63 (1 x 106 CFU). At 90 d after immunization, recovered lung cells were in vitro stimulated with Mtb extract for evaluation of Th1/Th2/Th17 cytokine profile by CBA in the culture supernatant. At day 91 after immunization, the animals were challenged with Mtb H37Rv (1 x 105 CFU) and 15 d after challenge (day 106), recovered lung cells were in vitro stimulated with Mtb extract and analyzed by immunophenotyping for determination of T-cell response (intracellular cytokine staining). Lungs were collected for histopathological analysis.