PMC5577078_01
The findings of nasal endoscopy and a biopsy. (A) There were necrotic lesions on the bilateral nasal mucosa (arrowheads). (B) Hematoxylin and Eosin staining shows the polymorphous tumor cells (magnification: 400 x). (C) Lymphoma cells expressing the NK cell marker CD56 were detected (CD56 stain, 400 x). (D) Staining for the T cell marker CD3 was positive (CD3 stain, 400 x).
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CD56-positive lymphoma cells (Fig. 1). Cytotoxic molecules, including granzyme B, perforin, and TIA-1, were detected, and EBV-encoded RNA (EBER) in situ hybridization was positive (Fig. 2); he was therefore diagnosed with ENKL.
The findings of in situ hybridization and immunohistochemistry. (A) EBV-encoded RNA (EBER) in situ hybridization shows that EBV-infected cells, including large numbers of copies of EBERs, were present in the tissue at 400x magnification. In addition, (B) granzyme B stain, (C) TIA-1 stain, and (D) perforin staining confirmed the diagnosis of nasal type, extranodal NK/T-cell lymphoma, (magnification: 400 x). EBER in situ hybridization was performed using the EBER PNA probe (Dako) and catalyzed signal amplification system (Dako). The commercially available primary antibodies used were as follows: granzyme B (mouse monoclonal, clone GrB-7, Dako), TIA-1 (mouse monoclonal, clone TIA-1, Coulter Immunology), and perforin (mouse monoclonal, clone 5B10, Novocastra).
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CD56-positive lymphoma cells (Fig. 1). Cytotoxic molecules, including granzyme B, perforin, and TIA-1, were detected, and EBV-encoded RNA (EBER) in situ hybridization was positive (Fig. 2); he was therefore diagnosed with ENKL.
Clinical course. With concurrent chemoradiotherapy, the renal function and urine abnormalities were improved. In addition, the PR3-ANCA titer was decreased gradually. PR3-ANCA: proteinase 3-antineutrophil cytoplasmic antibody
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CCRT gradually led to a reduction in the tumor size, improvement of the renal function and urine abnormalities, and a decline in the PR3-ANCA level (Fig. 3).
Immunohistochemical findings exploring the mechanism of PR3-ANCA production. (A) Neutrophils infiltrated in an angiocentric and angiodestructive growth pattern (MPO stain 400 x). (B) Apoptotic cells were seen (cleaved caspase 3 stain 400 x). (C) (D) Some neutrophils (arrowhead in C, D) were identified as apoptotic using serial sections (C; MPO stain 400 x, D; cleaved caspase 3 stain 400 x). (E) (F) The components of NETs, such as Cit-H3 and lactoferrin, were not detected (E; Cit-H3 stain 400 x, F; lactoferrin stain 400 x). The commercially available primary antibodies used were as follows: MPO (rabbit polyclonal, Dako), cleaved caspase 3 (rabbit polyclonal, Cell Signaling Technology), Cit-H3 (rabbit polyclonal, Abcam), and lactoferrin (rabbit polyclonal, GenWay Biotech). PR3-ANCA: proteinase 3-antineutrophil cytoplasmic antibody, MPO: myeloperoxidase, NETs: neutrophil extracellular traps, Cit-H3: citrullinated histone H3
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Infiltrated neutrophils were detected in an angiocentric and angiodestructive growth pattern by myeloperoxidase (MPO) staining (Fig. 4A). Some of them had become apoptotic (Fig. 4B-D). These stains revealed no formation of NETs (Fig. 4E, F).