Clonal VDJ Recombination of the Immunoglobulin Heavy Chain Gene by PCR in Classical Hodgkin’s Disease

Abstract

Although Hodgkin's disease (HD) has been a subject of much investigation, fundamental questions remain unanswered regarding its lineage and clonality. The authors used a polymerase chain reaction (PCR) technique to investigate whether clonal Variable-Diversity-Joining recombination of the immunoglobulin heavy (IgH) chain gene, a phenomenon that characterizes clonal B-cell proliferations, exists in nodular sclerosing (NSHD) and mixed cellularity (MCHD) Hodgkin's disease (so-called "classical" Hodgkin's disease). The isolation of DNA from paraffin-embedded tissue sections allowed for direct correlation of PCR results with the cell populations that were analyzed. Thirty-two cases were studied. These included 12 cases in which the Reed-Sternberg (RS) cells expressed the B-cell antigen, CD20, and 10 cases that were classified as syncytial variant of NSHD (3 CD20+, 7 B-cell antigen negative). Overall, clonal patterns of VDJ PCR products were found in 14 of 32 (44%) cases. These clonal patterns were identified in 7 of 12 (58%) cases of CD20+ classical HD and in 7 of 20 (35%) cases of B-antigen-negative classical HD. Clonal patterns were found in 3 of 10 cases of syncytial variant of NSHD, including 2 of 3 (67%) CD20+ cases and 1 of 7 (14%) B-cell antigen-negative cases. The results of this study provide support that a subset of HD represents a clonal B-cell neoplasm, and indicate that clonal IgH VDJ sequences are more frequently found in CD20+ HD.

PCR Assays Detect B-Lymphocyte Clonality in Formalin-Fixed, Paraffin-Embedded Specimens of Classical Hodgkin Lymphoma Without Microdissection

“…Only a handful of publications have described PCR-based clonality detection for HL using non-microdissected specimens 3, 5, 8-10, 4. Just 3 of these have assessed the use of the standardized BIOMED-2 primer system for FFPE HL specimens (Table 4).…”

IgH AND TcR-γ GENE REARRANGEMENTS IDENTIFIED IN HODGKIN'S DISEASE BY PCR DEMONSTRATE LACK OF CORRELATION BETWEEN GENOTYPE, PHENOTYPE, AND EPSTEIN-BARRVIRUS STATUS

“…Approximately 30 per cent of cases of classic HD showed IgH chain gene rearrangement, in overall agreement with reports using similar techniques but with a smaller number of cases. [5][6][7][8] Complete rearrangement of the IgH gene signifies commitment to B-lymphocyte lineage. 9 The study of TcR-gene rearrangements has practical implications, since TcRgenes have a limited V gene repertoire, despite extensive 23,[32][33][34] In addition, TcR-gene rearrangement occurs at an early stage of T-lymphocyte development and can be exploited as a marker of clonality in almost all T-lymphoid neoplasms.…”

“…3,4 The low detection rate of Ig and TcR gene rearrangements by Southern blotting has been attributed to either the paucity of tumour cells below the detection threshold or the actual absence of such gene rearrangements, leaving widely divergent possibilities for the derivation of HRS cells. With the use of the more sensitive and specific polymerase chain reaction (PCR), rearranged Ig heavy chain (IgH) genes have been detected recently in 25-50 per cent of classic HD (non-lymphocyte predominant subtypes), [5][6][7][8] supporting a clonal lymphocytic origin in a proportion of HD cases. While the current investigation on the study of HD by PCR using consensus primers was in progress, two groups confirmed the presence of Ig gene rearrangements in selected cases of HD in single HRS cells isolated by micromanipulation from frozen sections.…”

“…Example of PCR analysis of IgH chain gene rearrangement using Fr2a and J H primers in 12 cases of HD(1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12). 's are size markers; Bl (blank) is PCR amplification without a DNA template.…”

Hodgkin Disease Associated With T-Cell Non-Hodgkin Lymphomas

“…A substantial fraction of RS cells express lymphoid antigens; approximately 15% to 38% express CD20, and 11% to 24% express CD3. [27][28][29] Although early studies detected clonal immunoglobulin heavy chain gene rearrangements in RS cells in only 50% of cases, [27][28][29][30] individual cell micromanipulation and PCR have permitted the identification of clonal immunoglobulin heavy chain gene rearrangements in 90% to 95% of HD cases, with evidence of somatic mutation suggestive of germinal center origin. [31][32][33][34] Because most cases of HD are presumed to arise from the B-cell lineage, it is not surprising that a high incidence of other B-cell lymphomas is seen.…”