During ART, only frequencies of CD4⁺ cytokine-producing T cells specific to IRIS-associated Ags increase dramatically. PBMC samples from 5 IRIS patients (A) and 5 non-IRIS patients (B) were stimulated with MAC, H capsulatum, C neoformans, TB, CMV, JCV, or HIV-1. The IRIS-associated opportunistic infection and other known infecting pathogens tested for in the present assays are indicated for each patient. The number of stimulations performed with each PBMC sample was determined by the number of cells available, and priorities were given to those Ags to which a given patient was known to have been exposed. Gray boxes indicate the first time point within 3 months of clinical manifestation of IRIS. Dashed lines separate pre-ART from on-ART samples. Bold lines indicate T-cell responses to IRIS-associated Ags. IRIS patients were selected for illustration if stimulation data were available for at least 2 Ags and at least 4 time points. If 2 Ags fulfilled these criteria, all other stimulations with at least 2 data points were shown for that patient. Non-IRIS patients were selected according to the aforementioned criteria, as well as having known to be exposed to at least one of the Ags being tested: 1 indicates tp1; 2, tp2; 3, tp3; 4, tp4; and 5, tp5.