title: Multicolor flowcytometric immunophenotyping is a valuable tool for detection of intraocular lymphoma.
abstract: OBJECTIVE
  Intraocular lymphoma (IOL) is a rare condition and frequently difficult to distinguish from uveitis or other uveitis-masquerading syndromes. The diagnosis is confirmed by cytologic examination of ocular fluid specimens and more recently by molecular-immunoglobulin heavy chain (IGH) translocation or cytokine analysis. However, some of these more recent methods have not been validated by follow-up studies.
  
  
  METHODS
  Evaluation of a diagnostic test.
  
  
  METHODS
  In a cohort of 51 consecutive patients with a clinical suspicion of IOL, vitreous analysis was performed via multicolor flowcytometric immunophenotyping.
  
  
  METHODS
  Multicolor flowcytometric immunophenotyping was performed with CD45, CD3, CD19, CD20, anti-SmIgκ, and anti-SmIgλ antibodies. The presence of a clear B-cell population showing a disequilibrium of Igκ versus Igλ expression was used to confirm the diagnosis of non-Hodgkin lymphoma (NHL). Patients were followed for a minimum of 2 years (mean, 5.9 ± 2.0 years) to validate the accuracy of the method.
  
  
  METHODS
  The presence or absence of IOL during follow-up.
  
  
  RESULTS
  In 14 of 51 patients, a clinical diagnosis of IOL was confirmed using flowcytometric analysis. Of these 14 patients, 11 had primary IOL and 3 had metastasized secondary lymphomas. In 3 of 51 patients who were diagnosed with (central nervous system) NHL during follow-up, the test failed to confirm the presence of a clonal B-cell population. In 18 of the 34 other patients, an infectious or well-defined immunologic disorder was established during follow-up. The remaining 16 patients, with a minimal follow-up of 2 years, were diagnosed with idiopathic uveitis.
  
  
  CONCLUSIONS
  Multicolor flowcytometric analysis had 82.4% sensitivity and 100% specificity in patients with suspected IOL. This is comparable to the reported vitreous interleukin (IL)-6/IL-10 testing sensitivity of 0.8 and sensitivity of 0.65 to 0.95 by immunoglobulin heavy chain (IGH) gene arrangement testing in clinical cohorts. Because flowcytometric tests are readily performed in hematologic laboratories, this can be regarded as a useful method for confirming the clinical diagnosis of IOL.
  
  
  BACKGROUND
  The author(s) have no proprietary or commercial interest in any materials discussed in this article.
authors: Uveitis and Medical Retina Department, The Rotterdam Eye Hospital, Rotterdam, The Netherlands. t.missotten@oogziekenhuis.nl
authors: Tom
authors: T
authors: Missotten
authors: Dennis
authors: D
authors: Tielemans
authors: Jacolien E
authors: JE
authors: Bromberg
authors: P Martin
authors: PM
authors: van Hagen
authors: Ellen G
authors: EG
authors: van Lochem
authors: Jacques J M
authors: JJ
authors: van Dongen
authors: G Seerp
authors: GS
authors: Baarsma
authors: Anton W
authors: AW
authors: Langerak
mesh_terms: D000368:Aged; D015703:Antigens, CD / Q000032:analysis; D001402:B-Lymphocytes / Q000276:immunology; D015331:Cohort Studies; D005134:Eye Neoplasms / Q000175:diagnosis* / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D015321:Gene Rearrangement / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000378:metabolism; D016130:Immunophenotyping / Q000379:methods*; D008228:Lymphoma, Non-Hodgkin / Q000175:diagnosis* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D035583:Rare Diseases / Q000175:diagnosis* / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D015703:Antigens, CD; D007143:Immunoglobulin Heavy Chains