Figure 4.

Suppression under different stimulatory conditions. Flow plots showing CFSE-proliferation of CD4⁺ responder cells in suppression assays. (a) Comparison of CD3 microbeads stimulation versus MLR with dendritic cells. Data representative of two experiments. (b) Comparison of beads vs. plate-bound stimulation. CD15⁺ cells were isolated from PBMC of a patient with anxiety/depression, then used in suppression assay with responder PBMC (1:1 ratio), stimulated either by anti-CD3 microbeads (3.5 beads/cell, top), plate-bound anti-CD3 mAb (0.1 μg/mL, 2^nd and 3^rd rows), anti-CD3/28 microbeads (0.1 bead/cell, 4^th row) or plate-bound anti-CD3/28 (0.1 μg/mL of each, 5^th and 6^th rows). Additionally, aliquots of CD15⁺ cells were kept on ice for 3 hours prior to use in suppression assays (3^rd and 6^th rows). (c) Comparison of beads vs. soluble antibodies stimulation. CD4⁻ depleted cells from a lung transplant patient were used as suppressors, and responder cells were stimulated either with anti-CD3 microbeads (3.5 beads/cell, top), soluble anti-CD3 mAb (1 μg/mL, middle) or soluble anti-CD3/28 mAb (1 μg/mL, bottom). (d) Comparison of suppression by CD15⁺ cells isolated at room temperature vs. on ice. In total, 35 experiments with different types of stimulation and modification of CD15⁺ cells processing were performed, using PBMC from lung transplant (7), anxiety/depression (6), and lung cancer (3) patients, as well as healthy donors (6). Presented histograms and dot plots consist of 1,023 ± 73 (Mean ± SEM) events.