Frequency and phenotype of lipoarabinomannan (LAM)-responsive polycytotoxic T
cells in latent tuberculosis infection (LTBI) and posttuberculosis (post-TB)
donors. A total of 10 × 106 nonadherent
peripheral blood mononuclear cells (PBMCs) and autologous CD1+
antigen-presenting cells (APCs) (3:1 ratio) were stimulated with LAM (10
μg/ml) for 18 hours. Brefeldin was added for the final 4 hours of
incubation. Cells were then stained for CD3, IFN-γ, granulysin, granzyme
B, and perforin. At least 1 × 106 cells were
analyzed by flow cytometry. (A) A representative result for
LTBI donors (n = 28) and post-TB patients
(n = 51) is shown. (B) The individual
results of all donors investigated (solid circles, post-TB;
open circles, LTBI) are shown. Statistical significance was
calculated using the Mann–Whitney U test
(**P < 0.0001). Horizontal
bars represent the mean values. (C) The pie charts present a
summary of the individual data shown in (B).
(D) Nonadherent PBMCs and CD1+ APCs were
stimulated as described above, and
CD3+/IFN-γ+ polycytotoxic T cells were
further characterized by CD4 and CD8 labeling. At least
3 × 106 cells were acquired for each sample.
The graph presents one representative result of three different donors.
n.s. = not significant; SSC = side
scatter.