Figure 1
Figure 1. Histologic, immunohistologic, and cytogenetic features in a case of cyclin D1-negative lymphoma with CCND2-IGH fusion. (A,B) Hematoxylin and eosin–stained lymph node biopsy showing a vaguely nodular lymphoid infiltrate around an atrophic residual germinal center (A), composed of small cells with irregular nuclei admixed with scattered histiocytes (B). (C-F) Immunohistochemical findings: strong CD5 expression in the tumor cells around a negative residual germinal center (C); CD43 immunostaining of the lymphoma cells, with a lesser intensity than the reactive T cells (D); lack of cyclin D1 expression in the tumor cells; note that reactive histiocytes exhibit moderate nuclear staining (E); cyclin D2 nuclear expression in the tumor cells detected by a polyclonal anti-cyclin D2 antibody (Cell Signaling) (F). (G-I) FISH results on the bone marrow (G,H) and the lymph node (I) samples. Hybridization of spread metaphases with LSI IGH Dual Color, Break Apart Rearrangement probe (Vysis) shows one chromosome 14 with a normal hybridization pattern (juxtaposed centromeric orange and telomeric green probes) and one chromosome 14 hybridizing only with the centromeric probe, indicating an IGH break (arrow); loss of the telomeric part of the probe precluded identification of the partner chromosome (G). A break apart FISH assay for CCND2 locus rearrangement (telomeric BAC clone: RP11-578L13; centromeric BAC clone: RP11-388F6) shows 2 chromosomes 12 with a normal hybridization pattern and hybridization of the telomeric green probe to 14q, indicating a cryptic t(12;14)(p13:q32) (H). The CCND2 rearrangement is confirmed in the lymph node sample by interphase FISH as the nuclei show 2 yellow (normal chromosomes 12) and one or 2 green (derivative 14) signals with the 2 BAC clones (I). H&E and immunostains images were visualized under a Nikon Eclipse 80i microscope (Nikon, Tokyo, Japan) equipped with Nikon Plan Fluor 10×, 20×/0.50 NA, 40×/ objective lenses and a CFW-1310C camera (Scion, Frederick, MD); images were acquired using Histolab 5.131.1 (Alphelys, Plaisir, France) and processed using Adobe Photoshop v7.0. FISH images were acquired with a 100× immersion objective with an Olympus BX51 fluorescence microscope equipped with the appropriate filter sets, and were documented and processed using the FISH cytovision software.

Histologic, immunohistologic, and cytogenetic features in a case of cyclin D1-negative lymphoma with CCND2-IGH fusion. (A,B) Hematoxylin and eosin–stained lymph node biopsy showing a vaguely nodular lymphoid infiltrate around an atrophic residual germinal center (A), composed of small cells with irregular nuclei admixed with scattered histiocytes (B). (C-F) Immunohistochemical findings: strong CD5 expression in the tumor cells around a negative residual germinal center (C); CD43 immunostaining of the lymphoma cells, with a lesser intensity than the reactive T cells (D); lack of cyclin D1 expression in the tumor cells; note that reactive histiocytes exhibit moderate nuclear staining (E); cyclin D2 nuclear expression in the tumor cells detected by a polyclonal anti-cyclin D2 antibody (Cell Signaling) (F). (G-I) FISH results on the bone marrow (G,H) and the lymph node (I) samples. Hybridization of spread metaphases with LSI IGH Dual Color, Break Apart Rearrangement probe (Vysis) shows one chromosome 14 with a normal hybridization pattern (juxtaposed centromeric orange and telomeric green probes) and one chromosome 14 hybridizing only with the centromeric probe, indicating an IGH break (arrow); loss of the telomeric part of the probe precluded identification of the partner chromosome (G). A break apart FISH assay for CCND2 locus rearrangement (telomeric BAC clone: RP11-578L13; centromeric BAC clone: RP11-388F6) shows 2 chromosomes 12 with a normal hybridization pattern and hybridization of the telomeric green probe to 14q, indicating a cryptic t(12;14)(p13:q32) (H). The CCND2 rearrangement is confirmed in the lymph node sample by interphase FISH as the nuclei show 2 yellow (normal chromosomes 12) and one or 2 green (derivative 14) signals with the 2 BAC clones (I). H&E and immunostains images were visualized under a Nikon Eclipse 80i microscope (Nikon, Tokyo, Japan) equipped with Nikon Plan Fluor 10×, 20×/0.50 NA, 40×/ objective lenses and a CFW-1310C camera (Scion, Frederick, MD); images were acquired using Histolab 5.131.1 (Alphelys, Plaisir, France) and processed using Adobe Photoshop v7.0. FISH images were acquired with a 100× immersion objective with an Olympus BX51 fluorescence microscope equipped with the appropriate filter sets, and were documented and processed using the FISH cytovision software.

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