PMC3314557_01
Histological and immunohistochemical findings. (a) The testicular parenchyma shows no evidence of lymphomatous involvement (hematoxylin-eosin, original magnification x50). (b-c) Residual epididymal tubules are surrounded by a dense cellular infiltrate with a prevalent follicular architecture (hematoxylin-eosin, original magnification x100 and x630, respectively). The neoplastic cells strongly express CD20 (d) and CD10 (e) but are bcl-2 negative (f) (original magnification x250).
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F1
Microscopically, the testicular parenchyma showed no evidence of lymphomatous involvement (Figure 1a). The neoplasm predominantly displayed a follicular growth pattern (Figures 1b and 1c). Immunohistochemistry (IHC) investigation demonstrated that neoplastic cells strongly expressed CD45RB (LCA, clone 2B11+PD7/26, Dako), CD20 (clone L26, Dako), CD79a (clone JCB117, Dako), bcl-6 (clone PG-B6p, Dako) and CD10 (clone 56C6, Dako) (Figures 1d and 1e). On the other hand, staining for CD3 (clone F7.2.38, Dako), CD45RO (clone UCHL1, Dako), CD5 (clone 4C7, Thermo Scientific), CD23 (clone SP23, Dako) and bcl-2 (clone 124, Dako and clone 100/D5, Novocastra) were negative (Figure 1F).
Molecular findings. (a) Agarose gel electrophoresis to detect the rearrangement of the immunoglobulin heavy chain (IgH) rearrangement PCR products (semi-nested PCR). Lane 1: patient sample; lane 2: negative control (no DNA); lane 3: polyclonal control; lane 4: monoclonal control; lane MK: DNA ladder (501, 404, 331, 242, 190,147,111, 67, 34 bp fragments). (b) Agarose gel electrophoresis to detect BCL2/JH PCR products (nested PCR). Lane 1: negative control (no DNA); lane 2: patient sample; lane 3: BCL2 MBR positive control (plasmidic DNA containing BCL2/MBR rearrangement); lane MK: DNA ladder (501, 404, 331, 242, 190,147,111, 67, 34 bp fragments).
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F2
Finally, polymerase chain reaction (PCR) analysis revealed a monoclonal immunoglobulin heavy chain gene rearrangement and the detection of t(14;18)(IgH/BCL2) revealed the BCL/JH rearrangement (Figures 2a and 2b).