Extended Data Fig. 5: Cytokine response of human macrophages of donors with self-reported ancestry to Ghana, Cameroon, and Nigeria to distinct Mycobacterium tuberculosis complex strains.

This assay was conducted on cell culture supernatants collected from MDMs that were infected with 3 representative strains of L4 and L6, and also no infected MDMs. The infection was carried out with an MOI ~ 1:1, and the supernatants were collected at 24 and 96 hours post-infection (hpi). Three infection macrophage wells were tested per strain, time point, and donor. 13 human inflammatory cytokines-chemokines were screened using LEGENDplex. The production of six detected cytokines-chemokines is depicted in the figure, namely IL-1ß (a), TNF-α (b), MCP-1 (c), IL-6 (d), IL-8 (e) and IL-18 (f). The protein concentration in pg/mL was plotted on the y-axis, while the x-axis represented the controls (PBS and no-infected [NI] at 24 hpi and 96 hpi) and experimental conditions (infected with L4, and L6 at 24 hpi and 96 hpi). The stacked bars compared the mean production of these cytokines-chemokines at 24 hpi and 96 hpi within a lineage and across lineages. Each lineage is represented by three strains (three dots), and each strain comprises the averaged values of three donors. PBS, Macrophage Infection Media (MIM), and no-infected cells were used as controls. The MIM values were previously subtracted from no-infected and infected wells. The mean, standard error of the mean, and statistical results (ns, P >0.05; *P < 0.05; **P < 0.01; ***P < 0.001 and ****P < 0.0001) are depicted in the figures. Data were obtained from three independent infection experiments. The statistical results shown in the figures are two-sided p-values based on an unpaired t-test among both time points within the same lineage strain and on one-way ANOVA with Bonferroni post hoc test correction-multiple comparisons across distinct lineages at 24 hpi and 96 hpi. MDMs, Monocyte Blood Derived Macrophages; ns, not significant; NI, not-infected; L4, Lineage 4; L6, Lineage 6; CFU, Colony Forming Unit; MOI, Multiplicity of Infection; h, hours.

Source data