PCR

PCR-based clonality detection in angioimmunoblastic T-cell lymphoma

CLINICAL PRESENTATION, DIAGNOSIS, AND MANAGEMENT OF LYMPHOID MALIGNANCIES , , ,

Yu.V. Sidorova, Ye.Ye. Nikulina, N.G. Chernova, L.G. Gorenkova, Ye.A. Gilyazitdinova, S.K. Kravchenko, A.M. Kovrigina, and A.B. Sudarikov

Hematology Research Center, RF Ministry of Health, Moscow, Russian Federation

ABSTRACT

In this article, we discuss the issues of angioimmunoblastic T-cell lymphoma diagnosis, particularly the PCR-based methods of clonality detection. Assessments of T-and B-cell clonality was based on TCRG (Vg-Jg), TCRB (Vb-Jb, Db-Jb), IGH (FR1, FR2, and FR3), IGK (Vk-Jk, Vk/intron-Kde), or IGL (Vl-Jl) gene rearrangements in 15 patients. Clonal TCRG gene rearrangements were found in 66.7 % of primary biopsy samples. The combined use of primers for TCRG and TCRB gene rearrangements confirmed T-cell monoclonal population in most cases (86.7 %). The rate of B-cell clonality detection was 26.6 %. The presence of B-cell clones was not associated with monoclonal secretion in the blood or detecting Epstein-Barr virus positive B-cells in the biopsy samples. PCR-based clonality analysis is an important step in diagnosis of angioimmunoblastic T-cell lymphoma that enables identifying monoclonal origin of T-lymphocytes in most cases. However, when interpreting the results obtained by this method, it is necessary to consider the possibility of detecting B-cell monoclonal products of unclear origin.

Keywords: angioimmunoblastic T-cell lymphoma, PCR, clonality, T-cell antigen receptor

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Determination of B-cell clonality in Hodgkin’s lymphoma

EXPERIMENTAL STUDIES , , ,

Yu.V. Sidorova, N.V. Ryzhikova, S.Yu. Smirnova, E.E. Nikulina, B.V. Biderman, A.M. Kovrigina, T.N. Moiseeva, N.N. Sharkunov, and A.B. Sudarikov

Hematology Research Center, Moscow, Russian Federation

ABSTRACT

B-cell origin of Hodgkin’s lymphoma was demonstrated using microdissection and single cell PCR of Reed-Sternberg and Hodgkin cells (R. Kuppers et al., 1994). We assessed B-cell clonality in the biopsy samples of 35 patients with Hodgkin’s lymphoma without microdissection. B-cell clonality was evaluated using PCR amplification by IGH (FR1, FR2, FR3) and IGK (Vk-Jk, Vk/intron-Kde) gene rearrangements with multiplex BIOMED-2 primer sets and subsequent fragment analysis using ABI PRISM 3130 Genetic Analyzer (Applied Biosystems). Clonality was found in 11 out of 35 (31,5 %) formalin fixed paraffin-embedded (FFPE) lymph node specimens from patients with Hodgkin’s lymphoma. In 11 cases when both FFPE and fresh frozen samples were available, we observed the similar results with the specimens of both types. No correlation was found between the presence of B-cell clones and age, histological type of Hodgkin lymphoma, type of tumor cell growth (syncytial or diffuse), number of eosinophils in tissues, or CD20/CD15 expression on the surface of tumor cells. The high incidence of B-cell clonality determined in Hodgkin’s lymphoma biopsy samples makes the B-cell clonality assay unsuitable for differential diagnosis between Hodgkin’s lymphoma and B-cell lymphomas.

Keywords: Hodgkin’s lymphoma, B-cell clonality, PCR, immunoglobulin gene rearrangements.

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