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. 2008 Jun 12;118(7):2427–2437. doi: 10.1172/JCI35017

Figure 4. CLL tumor cells induce defective recruitment of signaling molecules to the immune synapse.

Figure 4

Healthy CD8+ and CD4+ T cells were cocultured (primary coculture) for 48 h in direct contact with either healthy allogeneic B cells or allogeneic CLL B cells and subsequently used in conjugation assays with sAg-pulsed third-party allogeneic healthy donor B cells (APCs, blue). T cell conjugates formed were analyzed by immunofluorescence and confocal microscopy (F-actin was stained red using rhodamine phalloidin). Images shown are representative of evaluation of 150 conjugates from 3 independent experiments stained green for (A) LFA-1, (B) high-affinity LFA-1 (mAb24), (C) Lck (5-min conjugation time), (D) TCR (nonpermeabilizing conditions), (E) Cdc42, (F) WASp, (G) filamin-A, and (H) dynamin-2. Quantitative image analysis of protein accumulation (green) at the immunological synapse is shown in Supplemental Figure 2. Arrows denote protein localization at the T cell–APC synapse site. Colocalization of proteins in the merged images is shown in yellow. Original magnification, ×63.