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. 1999 Aug 16;190(4):555–566. doi: 10.1084/jem.190.4.555

Figure 1.

Figure 1

Expression of CD80 and CD86 in allogeneic CD4+ T cell–EC cocultures. (A) Semiquantitative RT-PCR was performed on RNA samples harvested from resting CD4+ T cells, IFN-γ–treated HUVECs, or cocultures of CD4+ T cell with IFN-γ–treated HUVECs at various time points. Illustrated is one experiment representative of four. Resting CD4+ T cells occasionally show low level expression of CD86. (B) Western blot analysis of 24-h TNF-α–, soluble CD40 ligand–, and IFN-γ–activated ECs, or CD4+ T cells or 72-h CD4+ T cell–IFN-γ–treated EC cocultures. Two different anti-CD86 antibodies were used for our studies: IT2.2 (PharMingen; lanes 1–3) and B7-F3 (Bristol Myers Squibb; lanes 4–7). Illustrated are two representative blots showing absent CD86 expression on ECs and CD4+ T cells, but enhanced expression in CD4+–EC cocultures. Positive controls were CD86-transfected CHO cells. Data is representative of four experiments. (C) FACS® analysis of untreated HUVECs or 72-h IFN-γ (1,000 U/ml)–treated HUVECs for CD80, CD86, and HLA-DR (positive control). Control isotype mouse IgG was used as a negative control. As positive controls, we used anti-CD80 mAb and anti-CD86 mAbs bound to CD80- and CD86-transfected CHO cells, respectively (not shown).