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. 2007 Jan 18;8:2. doi: 10.1186/1471-2172-8-2

Figure 1.

Figure 1

Stimulation of LFA-1 through ICAM-2 and ICAM-3 in the presence of CD3/CD28 co-stimulation does not activate caspase-3. (A) Intracellular staining of cleaved-caspase 3 in cells treated with ICAM-1, ICAM-2, ICAM-3, CD3, CD3/CD28, CD3/CD28/ICAM-1, or CD3/CD28/ICAM-2, or CD3/CD28/ICAM-3. Naïve T cells were stimulated in 96-well coated plates and stained for CD4-PercpCy5.5 and intracellular cleaved-caspase-3-PE at 24 hrs. Data is representative of triplicate measures from one donor. Similar results were observed across three donors. (B) Cytometric bead array assessment of cleaved caspase-3 at 24, 48, and 72 hrs after naïve T cells were stimulated with 10, 100, 1000 ng of ICAM-1, ICAM-2, ICAM-3, CD3, CD3/CD28, ICAM-1/CD3/CD28, ICAM-2/CD3/CD28, or ICAM-3/CD3/CD28. Results are averaged from triplicate assays from three donors. Insert displays standard deviation for 48 hour measurement as denoted by the asterisk. (C) Absolute counts of cell divisions of naïve T cells stimulated with CD3/CD28 (control) or in the presence of TS1/22, ICAM-1, ICAM-2, or ICAM-3. T cells were labeled with CMFDA, stimulated in coated 96-well plates, and analyzed at day 7. TruCount beads were used to determine absolute viable cell numbers of divisional peaks detected by CMFDA fluorescence. Assays were performed on 6 individual donors. Triplicate measures from one donor are displayed in insert.