Article
Extended Data Fig. 12 | Determination of immune responses and BCG in
tissues 1 month after immunization. NHPs (cohort 6, n = 2) were immunized
with 5 × 10^7 BCG CFUs (IDhigh (a), IV (b), AE (c), or EB (d)); BCG CFUs and antigen-
responsive T cells were measured in various tissues 1 month later. Before
euthanasia, a fluorochrome-conjugated anti-CD45 antibody was injected
intravenously (ivCD45) such that circulating (intravascular) leukocytes were
uniformly stained (ivCD45+) while leukocytes in the tissue remained protected
from staining (ivCD45−). To investigate whether antigen-responsive (IFNγ+)
CD4 T cells were located in the ivCD45− lung tissue compartment, cells isolated
from lung lobes were re-stimulated in vitro with Mtb WCL and analysed by
intracellular cytokine staining. FACS plots show memory CD4 T cells in all
tissues collected from one of two NHPs per BCG regimen, organized by type/
location (systemic, peripheral LN, lung LN, BAL and lung lobes). The BAL and
lung responses from the IV BCG NHPs, shown in the bottom row of b, is
reproduced from Fig. 4b. Pre-infusion PBMC indicates PBMCs isolated from
whole blood collected just before anti-CD45 injection. Bar graphs show the
number of BCG CFUs in each respective tissue for each animal (colour-coded by
vaccine), if detected.