Figure 4. Immune responses and MTB bacterial counts in mice immunised with ICM.

Mice were immunised with 50 µg ICM (1∶20 antibody antigen ratio) or with Ag85B alone (30 µg), Ag85B+CT, BCG and PBS; two weeks after the final immunisation mice were either culled and their tissues (blood and spleens) used for immunological evaluation, or challenged i.n. with 70,000 MTB H37Rv. * Indicates statistically significant difference (p<0.05). A,B) Ag85B (A) and Acr (B) specific IgG, IgG1 and IgG2a serum responses determined by ELISA; shown are the mean values from 3 mice analysed in triplicates and in serial dilutions (indicated by differing patterns). C) Splenocyte proliferation after in vitro stimulation with Ag85B, measured by ³[H]-thymidine incorporation and expressed as stimulation indices (specific/nonspecific proliferation); n = 3 animals. D) IFN-γ release in splenocyte cultures (as in C) measured by an IFN-γ ELISA based kit. E) Lung bacterial counts in immunised mice; shown are the counts for individual mice (n = 6, except in some groups due to death of animals before the end of the experiment) and the means +/− SEM for each group.