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Review
. 2025 Jul 2;60(1):37.
doi: 10.1007/s44313-025-00088-0.

Effusion cytology of EBV-associated lymphoma: a concise review

Chih-Yi Liu  1   2 , Yen-Chuan Hsieh  3 , Sheng-Tsung Chang  4   5 , Hung-Chang Wu  6 , Shang-Wen Chen  7 , Shih-Sung Chuang  8
Affiliations
Review

Effusion cytology of EBV-associated lymphoma: a concise review

Chih-Yi Liu et al. Blood Res. .

Abstract

Epstein-Barr virus (EBV)-associated lymphomas can, on rare occasions, involve body cavities, making effusion cytology an important diagnostic tool. This mini-review explores the spectrum of EBV-related lymphomas that may be detected in serous fluids, including EBV-positive nodal T/NK-cell lymphoma (EBV + nT/NKCL), extranodal NK/T-cell lymphoma, primary effusion lymphoma, EBV-positive diffuse large B-cell lymphoma, and classic Hodgkin lymphoma. We present an index case of EBV + nT/NKCL with lymphomatous pleural effusion and discuss the cytologic features, differential diagnoses, and role of ancillary studies such as immunocytochemistry, EBER in situ hybridization, and molecular assays. Accurate diagnosis requires the integration of cytomorphologic, immunophenotypic, and molecular findings with clinical information to establish a definitive diagnosis and distinguish these aggressive lymphomas from reactive and non-hematologic mimics.

Keywords: Azurophilic granules; Cytotoxic phenotype; EBER; EBV; Effusion cytology; Immunophenotyping; T/NK-cell lymphoma.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: The study was conducted in accordance with the Declaration of Helsinki. The Institutional Review Board of Chi Mei Medical Center approved the study of the index case (protocol code CMH-IRB-2025–001, approval date: January 10, 2025). Consent for publication: The inform of consent has been waived by the Institutional Review Board of Chi Mei Medical Center. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Needle biopsy from a para-aortic lymph node. (A) Fragmented lymphoid tissue cores showing diffuse infiltration (H&E, × 40). (B) Sheets of large, atypical lymphocytes with vesicular nuclei, some with irregular nuclear contours and small nucleoli. Immunohistochemically, the tumor cells express CD2, CD3 (2C), TIA-1, and granzyme B (2E, GrB), but not CD4, CD5 (2D), CD8, CD20, CD138, TCR-BF1, or TCR-delta. (F) The tumor cells are diffusely positive for EBV by in situ hybridization (C-F, × 400)
Fig. 2
Fig. 2
Pleural effusion. (A) The effusion specimen is cellular with non-cohesive cells. (B, C) Singly scattered medium- to large-sized cells with vesicular nuclei, some indented, and one to three small nucleoli (A–C, Papanicolaou stain; A, × 400; B and C, × 1,000). (D–F) Tumor cells in the bone marrow aspirate smear (Liu stain; close-up view from × 1,000). (D) Two tumor cells and a benign histiocyte with hemophagocytosis. (E, F) Lymphoma cells show vesicular nuclei, small nucleoli, and a few azurophilic cytoplasmic granules
Fig. 3
Fig. 3
An example of HIV-negative primary effusion lymphoma from a 78-year-old man who presented with massive right pleural effusion (Panels A–E). He was negative for HIV infection. (A, B) Papanicolaou stain at × 400 and × 1,000 magnification, respectively, showing large neoplastic cells with vesicular nuclei, some containing a prominent central nucleolus. (C) Wright-Giemsa stain shows a neoplastic cell with plasmablastic features and a perinuclear hof (× 1,000). (D) Cell block section reveals large, non-cohesive tumor cells (H&E stain, × 1,000). (E) The tumor cells are positive for EBV by in situ hybridization (× 400). (F) An example of Burkitt lymphoma for comparison. Imprint cytology with Liu stain shows monomorphic neoplastic cells with round nuclei and small cytoplasmic lipid vacuoles (× 1,000). Lymphoglandular bodies are also present (arrow)
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