Figure 4. The constant of association Ka between monocytes and T cells (and T cell subsets) varies with the presence and nature of immune perturbations.

(A) Non-parametric spearman correlation between the frequency of T cell:monocyte complexes and the product of singlet T cells and monocyte frequencies in healthy subjects (n = 59). (B) Formula for the calculation of the T cell:monocyte constant of association Ka. T cell:monocyte complexes constant of association Ka in (C) active TB subjects at diagnosis and 2 months post treatment (n = 15), (D) individuals with acute dengue fever (n = 18), acute dengue hemorrhagic fever (n = 24) or previously infected (n = 47) and (E) previously vaccinated healthy adults (n = 16) before and three days post boost with Tdap vaccine, calculated as explained in B). (F) The constant of association Ka between monocytes and T cell subsets in active TB subjects at diagnosis (n = 25), individuals with acute dengue hemorrhagic fever (n = 24) and previously vaccinated healthy adults three days post boost with Tdap vaccine (n = 16), calculated as explained in B). Statistical differences over time and across cell populations within subjects were determined using the non-parametric paired Wilcoxon test; other statistical differences were determined using the non-parametric Mann-Whitney test; *, p<0.05; **, p<0.01; ***, p<0.01; ****, p<0.0001. Plots represent individual data points, median and interquartile range across all subjects within each cohort. Raw frequencies of T cell:monocyte complexes for the different disease cohorts are available on Figure 4—figure supplement 4. T cell:monocyte complexes were defined as the CD3+CD14+ cell population gated from live singlets as represented in Figure 1—figure supplement 2. CD4 and CD8 subsets within T cell:monocyte complexes were defined as presented in Figure 3E.

Figure 4—figure supplement 1. T cell:monocyte constant of association Ka in subjects with active TB, latent TB or TB uninfected individuals.

T cell:monocyte constant of association Ka was calculated as explained in Figure 4B from active TB samples (n = 15) collected at diagnosis from Sri Lanka, latent TB samples collected from subjects living in San Diego (n = 22) or Peru (n = 8), and TB uninfected samples collected from subjects living in San Diego (n = 29) or Sri Lanka (n = 14). Plots represent individual data points, median and interquartile range across all subjects within each cohort.

Figure 4—figure supplement 2. T cell:monocyte constant of association Ka fluctuates as a function of time following Tdap boost administration.

Previously vaccinated healthy subjects (n = 16) were re-immunized with Tdap and blood collected before, one day, three days, seven days and fourteen days post boost. Plots represent the median and interquartile range across all 16 subjects. T cell:monocyte constant of association Ka was calculated as explained in Figure 4B.

Figure 4—figure supplement 3. Comparison of constant of association Ka between monocytes and T cell subsets across different immune perturbations.

Constant of association Ka for each T cell subset and monocytes was calculated as explained in Figure 4B from active TB subjects at diagnosis (n = 25), individuals with acute dengue hemorrhagic fever (n = 24) and previously vaccinated healthy adults three days post boost with Tdap vaccine (n = 16). Plots represent individual data points, median and interquartile range across all subjects.

Figure 4—figure supplement 4. Frequencies of T cell:monocyte complexes in different immune perturbation models.

Frequencies of T cell:monocyte complexes (and T cell subsets:monocyte complexes) expressed as percent of live cells were determined in active TB subjects at diagnosis (n = 25) and two months post treatment (n = 15), individuals with acute dengue hemorrhagic fever (n = 24) and previously vaccinated healthy adults three days post boost with Tdap vaccine (n = 16). Statistical differences over time and across cell populations within subjects were determined using the non-parametric paired Wilcoxon test; other statistical differences were determined using the non-parametric Mann-Whitney test; *, p<0.05; **, p<0.01; ***, p<0.01; ****, p<0.0001. Plots represent individual data points, median and interquartile range across all subjects within each cohort. These frequencies were used to calculate the Ka values presented in Figure 4C–F, using the formula in Figure 4B.