Molecular Diagnostic Approach to Non-Hodgkin's Lymphoma

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Presentation transcript:

Molecular Diagnostic Approach to Non-Hodgkin's Lymphoma Daniel A. Arber The Journal of Molecular Diagnostics Volume 2, Issue 4, Pages 178-190 (November 2000) DOI: 10.1016/S1525-1578(10)60636-8 Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Immunoglobulin heavy chain gene rearrangement. Most PCR tests for this rearrangement use consensus primers directed against the framework three (FRIII) region and the heavy chain joining (JH or FRIV) region of the rearranged product. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Different methods for analyzing the immunoglobulin heavy chain PCR product are illustrated. A: A polyacrylamide gel illustrates both polyclonal and monoclonal results using FRIII/VLJH primers. Specimens 1–3 are run in duplicate and show a polyclonal pattern resulting in a smear pattern. Specimen 4 shows two reproducible, discrete bands. This biclonal pattern is considered evidence of a clonal population. Negative samples with, including a water control, a sample with no B lymphocytes (both with no amplifiable products), and a polyclonal B cell specimen (resulting in a smear pattern) are illustrated as lanes marked H2O, −, and −. A monoclonal B cell line control and a 1:100 dilution of that control are labeled + and 10−2. Both show a distinct band (arrow) of approximately 130 kb. MW lanes indicate molecular weight controls.B: The figure illustrates detection with a capillary electrophoresis instrument. Both demonstrate results of a monoclonal B cell population showing a large distinct peak, mixed with a polyclonal B cell population (multiple smaller peaks). In the upper portion, FRII/VLJH primers amplify a 243-kb clonal product; at bottom, FRIII/VLJH primers amplify an 82-kb clonal product. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 BCL-2/JH rearrangements usually involve the major breakpoint region (MBR) of the BCL-2 gene, but may also involve the minor cluster region (MCR) of the gene.BCL-1/JH rearrangements of t(11;14)(q13;q32) (not shown) rearrange in a similar fashion with the BCL-1 gene of chromosome region 11q13 fused 5′ to the JH region of the immunoglobulin heavy chain. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 A: Nuclear detection of BCL-1 (a.k.a. cyclin D1) protein overexpression by immunohistochemistry in mantle cell lymphoma is an excellent surrogate marker for the t(11;14) and reduces the need for the PCR detection method. B: ALK-1 immunohistochemistry is specific for abnormalities of the ALK gene in lymphoid neoplasms. C: In situ hybridization for EBER-1 RNA of the EBV demonstrates numerous EBV positive tumor cells in a case of nasal natural killer/T cell lymphoma. D: Some EBV-infected tumor cells, including the neoplastic cells of EBV-positive Hodgkin's disease, express the EBV latent membrane protein. Detection of this protein by immunohistochemistry is comparable to the in situhybridization method in those cases. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 FISH analysis for the t(8;14) of Burkitt's lymphoma may confirm this translocation (arrows) on metaphase spreads (left) or within intact nuclei (right), including nuclei from paraffin-embedded tissue (kindly provided by M. L. Slovak, Ph.D., City of Hope National Medical Center). The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 6 The T cell receptor γ chain locus on chromosome region 7p15 contains a limited number of variable and joining region genes that make it ideal for PCR amplification of the rearrangements. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 7 A diagnostic algorithm for clonality molecular testing in lymphoid proliferations. Additional studies could be performed to detect disease specific cytogenetic translocations. The Journal of Molecular Diagnostics 2000 2, 178-190DOI: (10.1016/S1525-1578(10)60636-8) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions