Figure 7. DLE immune activity is present in humans.

(A and B) Triplicate cultures of monocytes 1:1 with autologous CFSE-loaded CD3⁺ T cells in the presence of HKCA for 6 d from a Candida exposed donor (ED) who cohabitates with 2 patients with CMC or from random healthy volunteers (HV). On d 6, cells were stained and analyzed for CD3 and CD8 versus CFSE low (CFSE^lo) and CFSE high cells (A), and concentrations for indicated cytokines in the supernatants were measured (B). (C–H) hDLE was extracted from cellular lysates of PBMCs from ED or different random healthy volunteers. (C) d 3 IL-6 from 1e6 hDC^HKCA from unrelated healthy donors (n = 3) and cultured with 1e6 ceq of hDLE from ED or healthy volunteers. (D and E) 4e5 DC from a patient with STAT1 GOF mutation [proband (PB)], her affected daughter (AD), or healthy volunteers were pulsed with HKCA and cultured with 2e6 autologous PBMC and 1e5 ceq of hDLE to assess d 3 IL-1β, IL-6, and IL-10 (D) and IL-17A, IL-17F, and IL-22 (E). (F and G) d 3 IL-17A (F) and IL-10 (G) after 4e5 hDC^HKCA were cocultured with 2e6 autologous PBMCs with hDLE or murine HKCA:DLE before (Unsorted) or after bead enrichment for CD8 (CD8 Enr) or depletion (CD8 Dep). (H) IL-6 concentration for 1e6 hDC^HKCA was cultured 24 h with 1e5 ceq of hDLE or murine HKCA:DLE. Data shown are representative of 2 (A and B) or 3 or more (C–H) independent experiments using a different healthy volunteer per experiment with triplicate cultures and displayed means ± sem. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05, as determined by ANOVA.