These results thus confirmed that 1) small changes in the dose of antigen can significantly affect the efficacy of the vaccine, 2) increasing the vaccine dose beyond a specific threshold decreases the protective efficacy, 3) different animal models have different optimal doses (that probably depend on both the antigen and the adjuvant), and 4) the optimal dose of Ag85B-TB10.4 in IC31® does induce significant protection also in the guinea pig TB model. This evidence concerns the gene TMSB10 and tuberculosis.