PCR Analysis of Immunoglobulin Heavy Chain (IgH) and TcR-γ Chain Gene Rearrangements in the Diagnosis of Lymphoproliferative Disorders: Results of a Study of 525 Cases

Abstract

This report summarizes a cumulative 4-year experience in polymerase chain reaction (PCR) analysis of immunoglobin heavy chain (IgH) and TcR-␥ chain gene rearrangements in 525 cases of lymphoproliferative disorders. Because the sensitivity of the PCR methodology was found to be tissue dependent, in the study of the presence of clonal cell population in tissues containing a small number of polyclonal lymphocytes, such as skin and gastrointestinal biopsy specimens, we used the multiple-PCR run approach. In this latter methodology, we repeat the PCR reaction from the same sample at least three times to confirm the reproducibility of the results. In the study of 273 cases of B-or T-cell lymphomas with characteristic immunomorphological and clinical features, a clonal IgH or TcR-␥ chain gene rearrangement was detected in approximately 80% of cases. A clonal rearrangement involving both IgH and TcR-␥ chain genes was found in 10% of cases of both B-cell and T-cell lymphomas. The study of 167 cases of nonneoplastic lymphoid tissue samples showed the presence of clonally rearranged cell populations for IgH or TcR-␥ genes in 3 and 9% of cases, respectively. We also applied PCR for the study of 85 cases of lymphoproliferations with no definite diagnosis (i.e., benign versus malignant) after immunomorphological analysis. In 65 cases (76%), the correlation of immunomorphological features with the presence (48 cases) or the absence (17 cases) of clonal lymphoid cell populations led to a definite diagnosis. In almost all these cases, the final diagnosis was found to be in agreement with the clinical course. In the 20 remaining cases (24%), no definite diagnosis could be made. We also assessed the value of PCR in detecting bcl-2/J H gene rearrangement as an additional clonal marker in the diagnosis of follicular lymphoma. Bcl-2/J H rearrangement and/or IgH gene rearrangement was found in approximately 85% (71/85) of follicular lymphoma cases studied.

Immunoglobulin G is present in a wide variety of soft tissue tumors and correlates well with proliferation markers and tumor grades

“…2 The internal primers were FR3 and VLJH. 16 The PCR products were from 60 to 100 base pairs (bp) in size. The external primers for the VJ recombination of Igj were V1f, V3f, V7, and Jj1 through Jj4, as described previously.…”

A spindle cell variant of diffuse large B-cell lymphoma possesses genotypic and phenotypic markers characteristic of a germinal center B-cell origin

“…The upstream primers were complementary to the framework regions (FR1, FR2, or FR3) whereas the downstream primers were complementary to JH genes. 22 DNA samples were assayed in duplicate. Polymerase chain reaction products were analyzed by agarose gel electrophoresis and ethidium bromide staining.…”

Clonal Evolution of Gastric Lymphoma of Mucosa-Associated Lymphoid Tissue Type

“…But Ͼ40% of samples showed smears in PAGE by this method. Failure to produce bands with FR3A primer was reported previously by Theriault et al (16). They speculated that the failure to produce bands might be due to somatic mutations in MALT lymphoma.…”

“…For B-cell proliferation, rearrangement pattern of immunoglobulin heavy-chain gene was examined initially by the Southern blot method (10,11). Subsequently, polymerase chain reaction (PCR)-based methods were established and widely used to detect clones of cell in lymphoproliferative lesions (12)(13)(14)(15)(16). When compared with Southern blotting, this method is a faster, relatively simple, and inexpensive technique that avoids the use of radioactive materials and a requires smaller amount of DNA.…”