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PCR clonality detection in Hodgkin lymphoma.

Hebeda KM, Van Altena MC, Rombout P, Van Krieken JH, Groenen PJ - J Hematop (2009)

Bottom Line: A clonal population was found in 13 cases with the IGH FR1 and/or FR2/FR3 PCRs.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology 812, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands, k.hebeda@pathol.umcn.nl.

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.

No MeSH data available.


Related in: MedlinePlus

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)
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Fig1: 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)

Mentions: All lymph node biopsies were received fresh at the Department of Pathology where a part was collected for liquid nitrogen storage. The remaining tissue was formalin-fixed and paraffin-embedded. Four-micrometer tissue sections were stained with hematoxylin–eosin, and with monoclonal antibodies for CD20, CD30, CD79a (all DAKO), CD3 (Neomarkers), and CD15 (BD Diagnostics). A typical example is presented in Fig. 1. Staining was scored on the Hodgkin and Reed–Sternberg cells (HRS) cells: − <10%, +/− 10–50%, and + >50%. Average HRS cell percentages on HE and CD30 stains were independently scored by two observers (KH and HvK). In situ hybridization for Epstein–Barr virus (EBER) was performed on 22 cases according to the instructions of the manufacturer (DAKO). Immunophenotype and Epstein–Barr virus (EBV) status of the HRS cells is shown in Table 1. In addition, the percentage of small B cells in the background was estimated: − <5%, + 5–30%, and ++ >30%.Fig. 1


PCR clonality detection in Hodgkin lymphoma.

Hebeda KM, Van Altena MC, Rombout P, Van Krieken JH, Groenen PJ - J Hematop (2009)

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)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2713492&req=5

Fig1: 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)
Mentions: All lymph node biopsies were received fresh at the Department of Pathology where a part was collected for liquid nitrogen storage. The remaining tissue was formalin-fixed and paraffin-embedded. Four-micrometer tissue sections were stained with hematoxylin–eosin, and with monoclonal antibodies for CD20, CD30, CD79a (all DAKO), CD3 (Neomarkers), and CD15 (BD Diagnostics). A typical example is presented in Fig. 1. Staining was scored on the Hodgkin and Reed–Sternberg cells (HRS) cells: − <10%, +/− 10–50%, and + >50%. Average HRS cell percentages on HE and CD30 stains were independently scored by two observers (KH and HvK). In situ hybridization for Epstein–Barr virus (EBER) was performed on 22 cases according to the instructions of the manufacturer (DAKO). Immunophenotype and Epstein–Barr virus (EBV) status of the HRS cells is shown in Table 1. In addition, the percentage of small B cells in the background was estimated: − <5%, + 5–30%, and ++ >30%.Fig. 1

Bottom Line: A clonal population was found in 13 cases with the IGH FR1 and/or FR2/FR3 PCRs.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology 812, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands, k.hebeda@pathol.umcn.nl.

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.

No MeSH data available.


Related in: MedlinePlus