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Diagnostic vitrectomy 서울성모병원 안과 김규섭 primary intraocular lymphoma : masquerade syndrome diagnostic delay (11-21.4 months) - 60-80% : CNS tumor within 29 months - poor Px : 80% died within 2 years *** Clinical suspicion of intraocular lymphoma : most important - severe vitritis and poor VA –> vitrectomy 2009 Ocular Immunology & Inflammation Diagnositc vitrectomy • lymphoma cell : fragile and easily degenerate in the vitreous -> appropriate handling and processing : critical -> ocular pathologist involvement : essential • stop or taper steroid before vitrectomy : for better diagnostic yield • Obtaining - undiluted vitreous (1-2 mL) - diluted vitreous (3-5 mL) • Handling - cell culture medium (RPMI, Roswell Park Memorial Institute) or normal saline or direct transfer - sent immediately to an awaiting cytologist - Importantly, alcohol fixation may jeopardize the identification of PIOL cells in the vitreous sample. No fixative is required. <Diagnostic testing> • undiluted vitreous (1-2 mL) -> cytocentrifuge - cells : cytology, immunohistochemistry - supernatant : cytokine analysis, molecular analysis with PCR • diluted vitreous (3-4 mL) - flow cytometry - bacterial, funagl culture <Diagnostic testing> • Cytology : gold standard - atypical lymphoma cell 확인 (mostly B cell origin) ** The monoclonality supports the cytological diagnosis of lymphoma • Immunohistochemistry - stain cell surface marker - differentiate B- (CD19, 20…) and T-cell (CD3…) origin • Flow cytometry - analyze several different markers - require fewer cells than slide-based immunohistochemistry • microdissection and PCR - DNA extracted from B-lymphocyte is amplified with PCR - detecting clonal rearrangement of the Ig heavy chain (CDR3) • Cytokine analysis - IL 10 (malignant B-cell) / IL-6 (Inflammatory cells) ratio > 1.0 (suggestive of PIOL) * Chorioretinal Bx: - lymphoma cell: typically between RPE and Bruch’s memb may invade vitreous - Superficial : necrotic - Deep : viable 2009 Ocular Immunology & Inflammation