Figure 1.
Figure 1. Immunohistochemical analysis of C/EBPβ expression in normal lymphoid tissue and in different lymphoid neoplasias. (A) C/EBPβ protein expression in normal lymphoid tissue (tonsil). In the germinal center, mantle zone, and interfollicular T-cell area, C/EBPβ is strongly expressed in the nuclei of scattered cells, whereas most surface epithelial cells express high levels of C/EBPβ. Immunoperoxidase (IP) stain, 100×. C/EBPβ-positive cells within the germinal center correspond to follicular dendritic cells (inset; IP stain, 400×). (B) Double staining for C/EBPβ (brown) and clusterin (purple) reveals that the C/EBPβ-positive germinal center cells express the dendritic cell marker clusterin. Note also that the clusterin stain highlights the dendritic cell processes. IP stain, 650×. (C-D) Double staining for C/EBPβ (brown) and CD3 (purple) (C) or CD20 (purple) (D) in the germinal center demonstrates that the CD3+ T cells and CD20+ B cells are negative for C/EBPβ. IP stain, 650×. (E) C/EBPβ expression in mantle cell lymphoma. The neoplastic cells are negative, whereas the intratumoral histiocytes are positive and serve as an internal positive control. IP stain, 100×. (F-G) C/EBPβ in 2 cases of classic Hodgkin lymphoma. Faint nuclear reactivity is seen in the Reed-Sternberg cells in 1 case (G), whereas the second case (F) shows no C/EBPβ expression. In both cases, abundant histiocytes are strongly positive for C/EBPβ. IP stain, 650×. (H) C/EBPβ expression in unspecified PTCL. Neoplastic cells are negative, whereas intratumoral histiocytes are positive. IP stain, 100×. (I-J) C/EBPβ expression in a second case of unspecified PTCL. (I) Double staining for C/EBPβ (brown) and CD3 (purple) demonstrates that a small population of the neoplastic T cells (arrows) shows weak nuclear staining for C/EBPβ. In contrast, compare the strong nuclear positivity of the reactive histiocytes. IP stain, 400×. (J) Double staining for C/EBPβ (brown) and CD68 (purple) confirms the histiocytic origin of the strong C/EBPβ-positive cells. IP stain, 650×. (K) Unspecified PTCL with abundant histiocytes. Double staining for C/EBPβ (brown) and CD3 (purple) demonstrates that C/EBPβ is not expressed in the CD3+ tumor cells. IP stain, 100×. (L) C/EBPβ expression in ALCL, ALK-negative, with weak nuclear staining in tumor cells. In contrast, reactive histiocytes showed strong nuclear positivity with C/EBPβ. IP stain, 250×. Images were acquired using a Hitachi camera HW/C20 (Hitachi, Tokyo, Japan) installed in a Zeiss Axioplan microscope (Zeiss, Jena, Germany) using Intellicam software. Plan-Neofluar 10×/0.30 numeric aperture (NA), 20×/0.50 NA, and 40×/0.75 NA objectives as well as a Plan-Apochromat 63×/1.40 oil objective were used. Images were processed using Adobe Photoshop (Adobe Systems, San Jose, CA).

Immunohistochemical analysis of C/EBPβ expression in normal lymphoid tissue and in different lymphoid neoplasias. (A) C/EBPβ protein expression in normal lymphoid tissue (tonsil). In the germinal center, mantle zone, and interfollicular T-cell area, C/EBPβ is strongly expressed in the nuclei of scattered cells, whereas most surface epithelial cells express high levels of C/EBPβ. Immunoperoxidase (IP) stain, 100×. C/EBPβ-positive cells within the germinal center correspond to follicular dendritic cells (inset; IP stain, 400×). (B) Double staining for C/EBPβ (brown) and clusterin (purple) reveals that the C/EBPβ-positive germinal center cells express the dendritic cell marker clusterin. Note also that the clusterin stain highlights the dendritic cell processes. IP stain, 650×. (C-D) Double staining for C/EBPβ (brown) and CD3 (purple) (C) or CD20 (purple) (D) in the germinal center demonstrates that the CD3+ T cells and CD20+ B cells are negative for C/EBPβ. IP stain, 650×. (E) C/EBPβ expression in mantle cell lymphoma. The neoplastic cells are negative, whereas the intratumoral histiocytes are positive and serve as an internal positive control. IP stain, 100×. (F-G) C/EBPβ in 2 cases of classic Hodgkin lymphoma. Faint nuclear reactivity is seen in the Reed-Sternberg cells in 1 case (G), whereas the second case (F) shows no C/EBPβ expression. In both cases, abundant histiocytes are strongly positive for C/EBPβ. IP stain, 650×. (H) C/EBPβ expression in unspecified PTCL. Neoplastic cells are negative, whereas intratumoral histiocytes are positive. IP stain, 100×. (I-J) C/EBPβ expression in a second case of unspecified PTCL. (I) Double staining for C/EBPβ (brown) and CD3 (purple) demonstrates that a small population of the neoplastic T cells (arrows) shows weak nuclear staining for C/EBPβ. In contrast, compare the strong nuclear positivity of the reactive histiocytes. IP stain, 400×. (J) Double staining for C/EBPβ (brown) and CD68 (purple) confirms the histiocytic origin of the strong C/EBPβ-positive cells. IP stain, 650×. (K) Unspecified PTCL with abundant histiocytes. Double staining for C/EBPβ (brown) and CD3 (purple) demonstrates that C/EBPβ is not expressed in the CD3+ tumor cells. IP stain, 100×. (L) C/EBPβ expression in ALCL, ALK-negative, with weak nuclear staining in tumor cells. In contrast, reactive histiocytes showed strong nuclear positivity with C/EBPβ. IP stain, 250×. Images were acquired using a Hitachi camera HW/C20 (Hitachi, Tokyo, Japan) installed in a Zeiss Axioplan microscope (Zeiss, Jena, Germany) using Intellicam software. Plan-Neofluar 10×/0.30 numeric aperture (NA), 20×/0.50 NA, and 40×/0.75 NA objectives as well as a Plan-Apochromat 63×/1.40 oil objective were used. Images were processed using Adobe Photoshop (Adobe Systems, San Jose, CA).

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