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Updated: Aug 11, 2026
VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
Analysis of clonal immunoglobulin heavy chain rearrangements in ocular lymphoma
Joachim M Baehring1, Sofia Androudi, Janina J Longtine
1Brain Tumor Center, Department of Neurology, Massachusetts General Hospital, 100 Blossom Street, Boston, MA 02114, USA.
Insights
Analysis of immunoglobulin heavy chain gene rearrangements (AIGHR) improved intraocular lymphoma (IOL) diagnosis. This molecular test, when combined with traditional methods, increased diagnostic accuracy for IOL, aiding in challenging cases.
Area of Science:
- Ophthalmology
- Oncology
- Molecular Biology
Background:
- Intraocular lymphoma (IOL) diagnosis is challenging due to paucicellular vitreous samples.
- Traditional methods like cytopathology and flow cytometry have limitations in sensitivity for IOL detection.
- Immunoglobulin heavy chain (IgH) gene rearrangement analysis (AIGHR) is a valuable tool in systemic lymphoma but its utility in IOL is not well-established.
Purpose of the Study:
- To evaluate the sensitivity and specificity of AIGHR as an adjunct diagnostic tool for intraocular lymphoma (IOL).
- To assess the diagnostic yield of AIGHR compared to cytopathology and flow cytometry in patients with suspected IOL.
Main Methods:
- Retrospective review of clinical records for patients undergoing pars plana vitrectomy (PPV) for suspected IOL.
- Analysis of immunoglobulin heavy chain (IgH) gene rearrangements (AIGHR) using seminested polymerase chain reaction (PCR) on vitreous fluid.
- Comparison of AIGHR results with cytopathology and flow cytometry findings.
Main Results:
- AIGHR demonstrated a specificity of 1.0 and a sensitivity of 0.64 in diagnosing IOL.
- In two cases, AIGHR identified lymphoma when cytopathology or flow cytometry were negative.
- Clonal IgH gene rearrangements were detected in four specimens initially classified as lymphoma-negative by conventional methods.
Conclusions:
- AIGHR significantly supplements cytopathology and flow cytometry for intraocular lymphoma (IOL) diagnosis.
- The integration of AIGHR enhances the diagnostic yield and accuracy for IOL.
- AIGHR is a valuable molecular adjunct for diagnosing intraocular lymphoma.
Background:
The morphologic diagnosis of primary and metastatic intraocular lymphoma (IOL) was made difficult by the paucicellular specimens with fragile populations of lymphocytes retrieved through pars plana vitrectomy (PPV). The analysis of immunoglobulin heavy chain (IgH) gene rearrangements (AIGHR) was used as an adjunct to cytopathology and flow cytometry in systemic lymphoma. In IOL, the sensitivity and specificity of AIGHR are unknown.
Methods:
The authors reviewed the clinical records of patients who underwent PPV for suspicion of IOL at the Immunology and Uveitis Service of the Massachusetts Eye and Ear Infirmary (Boston, MA) between 2000 and 2002. AIGHR was performed as a routine diagnostic test on cell lysates isolated from < 0.5 mL of vitreous fluid. The authors used seminested polymerase chain reaction (PCR) with consensus primers for the VDJ-region of the IgH gene. PCR products were analyzed by polyacrylamide gel electrophoresis.
Results:
Thirty patients (37 specimens) with chronic vitritis and 17 patients (23 specimens) with IOL were included. The specificity of vitreous fluid cytopathology, flow cytometry, and AIGHR was 1.0, and the sensitivity values were 0.24, 0.36, and 0.64, respectively. AIGHR was negative in two patients for whom cytopathology or flow cytometry revealed the diagnosis of lymphoma. Clonal IGHR was found in four specimens classified as negative for lymphoma based on cytopathology and flow cytometry.
Conclusions:
AIGHR supplemented cytopathology and flow cytometry to increase the diagnostic yield in IOL.
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