Research Report 2010 - MDC

ABGene deregulation and its impact on formation of translocations in anaplastic large cell lymphoma (ALCL). (A) Immunohistchemistry of Fra2and Id2 in ALCL carrying (left) or lacking (right) t(2;5). Cells with positive signals are stained in red. Large neoplastic cells show a strong nuclearstaining for Fra2 and Id2, whereas no (Fra2) or only weak (Id2) signals are detectable in the small non-neoplastic cells. (B) Schematic view of thebreakpoint proximal genes up-regulated in ALCL cells showing the localization of FRA2 (located on 2p23), ID2 (2p25), and CSF1R (5q33.1). In thecase of t(2;5), the fusion of NPM (5q35) and ALK (2p23) results in the NPM-ALK fusion gene. Colored bars indicate positions of probes used forFISH in Mathas et al., PNAS, 2009.Activation-dependent intracellular redistribution of EBAG9 toward the immunological synapseEBAG9 (green) relocalizes toward the immunological synapse when cytotoxic T lymphocytes are conjugated withCD3/CD28 coated microbeads, which act as surrogate antigen-presenting cells (opaque structures).CTLs were mixed with CD3/CD28 coated microbeads, incubated for 5 min at 37°C, plated on coverslips, and incubatedfor another 30 min before fixation and permeabilization. Lytic granules were stained with anti-Cathepsin D antibody(red). A non-conjugated and non-polarized T cell is shown in the top row. Polarized T cells with conjugated beads areshown below. Images were obtained with a Zeiss LSM 510 confocal microscope setup. Scale bars, 5 micron. Insets haveadditional magnifications of 2- to 3-fold.128 Cancer Research