Multicolor Flowcytometric Immunophenotyping Is a Valuable Tool for Detection of Intraocular Lymphoma

doi:10.1016/j.ophtha.2012.11.007

Ophthalmology

Volume 120, Issue 5, May 2013, Pages 991-996

https://doi.org/10.1016/j.ophtha.2012.11.007 Get rights and content

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.

Design

Evaluation of a diagnostic test.

Participants

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.

Main Outcome Measures

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.

Financial Disclosure(s)

The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Section snippets

Patients

Fifty-nine consecutive vitreous samples from 51 patients presenting between 2002 and 2007 with a (pseudo)uveitis that was clinically suspected to be an IOL were retrospectively analyzed. These (pseudo)uveitis cases included patients with known CNS lymphoma, systemic lymphoma, or noninfectious uveitis refractory to high-dose steroids and patients with clinically suspected choroidal infiltrates. After informed consent to use their clinical data for follow-up purposes and quality analysis, all

Results

In the group of 51 patients (23 men and 28 women), 10 (20%) had a history of extraocular NHL and 41 (80%) had a clinical presentation compatible with IOL or concerned uveitis cases refractory to anti-inflammatory treatment. The mean ± SD age at the time of first ocular symptoms was 65.4±13.3 years and 66.7±13.0 years at the time of vitreous sampling and diagnosis, respectively.

Discussion

Cytomorphologic examination of vitreous samples is frequently inconclusive because of the limited number of cells contained in them, especially if the patient is treated with corticosteroids before the vitreous biopsy. False-negative results of up to 30% were reported in clinical cohorts.² Several additional tests have been proposed in recent years. Interleukin-10 measurements alone or the IL-10/IL-6 ratio was validated by retrospective analysis of samples from patients with known primary IOL.

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: Manuscript no. 2011-1481.

: Financial Disclosure(s): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

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Original articleMulticolor Flowcytometric Immunophenotyping Is a Valuable Tool for Detection of Intraocular Lymphoma

Objective

Design

Participants

Methods

Main Outcome Measures

Results

Conclusions

Financial Disclosure(s)

Section snippets

Patients

Results

Discussion

Ophthalmology

Ophthalmology

Am J Ophthalmol

Ophthalmology

J Mol Diagn

Blood

Surv Ophthalmol

Ocular and central nervous system lymphoma: clinical features and diagnosis

Ocul Immunol Inflamm

Association of interleukin 10 in the vitreous and cerebrospinal fluid and primary central nervous system lymphoma

Arch Ophthalmol

Interleukin-10 and intraocular-central nervous system lymphoma [letter]

Ophthalmology

IL-10 measurement in aqueous humor for screening patients with suspicion of primary intraocular lymphoma

Invest Ophthalmol Vis Sci

Elevated vitreous interleukin-10 level is not diagnostic of intraocular-central nervous system lymphoma

Ophthalmology

Original article
Multicolor Flowcytometric Immunophenotyping Is a Valuable Tool for Detection of Intraocular Lymphoma