Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Mar;2(1):34-41.
doi: 10.1007/s12308-009-0024-1. Epub 2009 Feb 20.

PCR clonality detection in Hodgkin lymphoma

K M Hebeda  1 , M C Van Altena, P Rombout, J H J M Van Krieken, P J T A Groenen
Affiliations

PCR clonality detection in Hodgkin lymphoma

K M Hebeda et al. J Hematop. 2009 Mar.

Abstract

B-cell clonality detection in whole tissue is considered indicative of B-cell non-Hodgkin lymphoma (NHL). We tested frozen tissue of 24 classical Hodgkin lymphomas (cHL) with a varying tumor cell load with the multiplex polymerase chain reaction (PCR) primer sets for IGH and IGK gene rearrangement (BIOMED-2). A clonal population was found in 13 cases with the IGH FR1 and/or FR2/FR3 PCRs. Using the IGK-VJ and IGK-DE PCRs, an additional six cases had a dominant clonal cell population, resulting in a detection rate of 79% in frozen tissue. Of 12 cases, also the formalin-fixed and paraffin-embedded (FFPE) tissue was tested. Surprisingly, in eight of the 12 FFPE cases with acceptable DNA quality (allowing PCR amplification of >200 nt fragments), the IGK multiplex PCRs performed better in detecting clonality (six out of eight clonal IGK rearrangements) than the IGH PCRs (four out of nine clonal rearrangements), despite a rather large amplicon size. There was no evidence of B-cell lymphoma during follow-up of 1 to 6 years and no correlation was found between the presence of a clonal result and Epstein-Barr virus in the tumor cells. Our results indicate that the present routine PCR methods are sensitive enough to detect small numbers of malignant cells in cHL. Therefore, the presence of a clonal B-cell population does not differentiate between cHL and NHL.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Example of nodular sclerosing classical Hodgkin lymphoma (patient 8). a HE stain. The HRS cells, indicated by arrows, are positive for CD30 (b) and CD15 (c). Stains for CD20 (d) and CD3 (e) are negative on the HRS cells (×20)
Fig. 2
Fig. 2
Clonality assessment. GeneScan results of the gene rearrangement profile of a cHL frozen biopsy sample (patient 12) showing clonal Ig rearrangements with multiple primer sets in a polyclonal background of B cells (a) and a polyclonal tonsil biopsy control (b)
Fig. 3
Fig. 3
Examples of PCR on DNA samples from corresponding frozen and paraffin-embedded tissue. All examples show clonal products and a polyclonal background. (a) Case 1: the IGH-V(D)J rearrangement with the FR3 and FR1 amplicon sizes of 177 and 371 nt is clearly detectable in the DNA from frozen tissue, but not in the DNA sample from paraffin-embedded tissue. Case 14 (b) and case 12 (c), IGK-DE amplicon sizes of 374 and 274 nt are detected in frozen and paraffin-embedded tissue. DNA obtained from a tonsil is used as the polyclonal control sample in each PCR experiment
See this image and copyright information in PMC

References

    1. Tamaru J, Hummel M, Zemlin M, Kalvelage B, Stein H. Hodgkin's disease with a B-cell phenotype often shows a VDJ rearrangement and somatic mutations in the VH genes. Blood. 1994;84(3):708–15. - PubMed
    1. Evans PA, Ch Pott, Groenen PJ, Salles G, Davi F, Berger F, Garcia JF, Krieken JH, Pals S, Kluin P, Schuuring E, Spaargaren M, Boone E, Gonzalez D, Martinez B, Villuendas R, Gameiro P, Diss TC, Mills K, Morgan GJ, Carter GI, Milner BJ, Pearson D, Hummel M, Jung W, Ott M, Canioni D, Beldjord K, Bastard C, Delfau-Larue MH, Dongen JJ, Molina TJ, Cabecadas J. Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia. 2007;21(2):207–14. doi: 10.1038/sj.leu.2404479. - DOI - PubMed
    1. Chute DJ, Cousar JB, Mahadevan MS, Siegrist KA, Silverman LM, Stoler MH. Detection of immunoglobulin heavy chain gene rearrangements in classic Hodgkin lymphoma using commercially available BIOMED-2 primers. Diagn Mol Pathol. 2008;17(2):65–72. doi: 10.1097/PDM.0b013e318150d695. - DOI - PubMed
    1. McClure RF, Kaur P, Pagel E, Ouillette PD, Holtegaard CE, Treptow CL, Kurtin PJ. Validation of immunoglobulin gene rearrangement detection by PCR using commercially available BIOMED-2 primers. Leukemia. 2006;20(1):176–9. doi: 10.1038/sj.leu.2404049. - DOI - PubMed
    1. Dongen JJ, Langerak AW, Bruggemann M, Evans PA, Hummel M, Lavender FL, Delabesse E, Davi F, Schuuring E, Garcia-Sanz R, Krieken JH, Droese J, Gonzalez D, Bastard C, White HE, Spaargaren M, Gonzalez M, Parreira A, Smith JL, Morgan GJ, Kneba M, Macintyre EA. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia. 2003;17(12):2257–317. doi: 10.1038/sj.leu.2403202. - DOI - PubMed