Detection of concurrent/recurrent non-hodgkin's lymphoma in effusions by PCR

Abstract

A subset of patients with non-Hodgkin's lymphoma (NHL), present with or subsequently develop lymphocytic effusions. Differential diagnosis between reactive lymphocytosis and recurrent low-grade NHL is difficult by cytology alone. We studied the use of polymerase chain reaction (PCR)-based techniques to detect concurrent/recurrent NHL. Both primary tumors and atypical lymphocytic effusions of 12 low-grade B-NHL patients and 4 T-NHL patients were studied. Six pleural effusions (reacitve/carcinomatous), in patients with no history of NHL, were included. Samples were amplified by PCR, using Fr3, Fr2, LJH, and VLJH primers specific for the immunoglobulin heavy chain (IgH) gene and Vγ-8, Vγ9, Vγ10, Vγ11 and Jγ1/Jγ2 consensus primers specific for the T-cell receptor gamma (TCR-γ) gene. IgH gene PCR products were analyzed by polyacrylamide gel electrophoresis (PAGE). TCR-γ gene PCR products were analyzed using a novel nonradioactive single-stranded conformational polymorphism (SSCP) procedure. IgH gene rearrangement analysis demonstrated monoclonality in 11 12 primary low-grade B-NHLs. Idential monoclonal bands were found in both primary tumor and effusion in 9 patients. TCR-γ gene rearrangement analysis demonstrated monoclonality in 4 of 4 primary T-NHLs. Identical monoclonal banded patterns were found in both primary tumor and effusion in 3 patients. Our results strongly support the diagnosis of concurrent/recurrent NHL in 13 of 16 (81%) cases of atypical lymphocytic effusions. IgH/PAGE and TCR-γ/SSCP analyses are useful tools in the diagnoses of lymphocytic effusions in patients with NHL.