Reduced cytokine and chemokine responses in HSV-2–infected macaque DC-stimulated T-cell cocultures. MLR cultures were set up with uninfected (–) and HSV-2–infected (+) immature macaque moDCs (1 DC to 40 T cells), as described in Figure 5. Cell-free supernatants were collected and assayed (22-plex Luminex assay) prior to addition of 3H-TdR on day 5. Additional MLR cocultures were set up to assay IL-2 after 1 day of culture. The mean concentration (pg/mL) of each cytokine/chemokine (± SEM) is shown from 3 to 7 experiments using DCs and T cells from different donors. There is no error bar in the CCL3 graph because all values were above the highest limit of detection for the assay and have been set as this maximal value for each. Samples from the DC (5 × 104 cells per well) and T-cell (2 × 105 cells per well) controls cultured in medium were also measured. T cells produced negligible amounts of these factors when cultured alone. The amounts of factors released by the control DC cultures were close to or below the low detection limit for all except CXCL8. Even the 5 × 104 DCs per well released detectable amounts of CXCL8, but these were typically 1.5- to 2.2-fold less than that released in the DC–T cell cocultures (results from 4 experiments). Comparable results were obtained with human DCs from 2 separate donors, confirming that HSV-2–infected human DCs induced lower cytokine/chemokine responses in the allogeneic MLR.
