Hodgkin and Reed–Sternberg cells in lymphocyte predominant Hodgkin disease represent clonal populations of germinal center-derived tumor B cells

Abstract

Among the four subtypes of Hodgkin disease (HD), lymphocyte-predominant (LP) HD is now generally considered as a separate entity. The B cell nature of the typical Hodgkin and Reed–Sternberg (HRS) cells and their variants (L and H, lymphocytic and histiocytic cells) in LP HD has long been suspected, but the question of whether these cells represent a true tumor clone is unclear. We previously demonstrated clonal Ig gene rearrangements in one case of LP HD. In the present study, five cases of LP HD were analyzed by micromanipulation of single HRS cells from frozen tissue sections and DNA amplification of rearranged Ig heavy chain genes from those cells. Clonal V gene rearrangements harboring somatic mutations were detected in each case. In three cases ongoing somatic mutation was evident. This shows that HRS cells in LP HD are a clonal tumor population derived from germinal center B cells. The pattern of somatic mutation indicates that HRS cells in LP HD are selected for antibody expression. This, and the presence of ongoing mutation discriminates LP from classical HD.

B-cell development in progressively transformed germinal centers: similarities and differences compared with classical germinal centers and lymphocyte-predominant Hodgkin disease

“…10,16,17 Consequently, to study the relation between LPHD and PTGCs, we analyzed, from each of 2 lymph nodes, Ki67 ϩ cells from 2 individual PTGCs for clonal expansion across the borders of individual PTGCs.…”

“…1,2,26,27,[32][33][34] In LPHD, a single clone of lymphocytic and histiocytic cells, characterized as the descendant of a centroblast, is found in the different nodules of the lymphoma. 10,16,17 Although the detection of single clones of Ki67 ϩ cells encompassing 50% to 70% of the mutated Ki67 ϩ cells in 3 PTGCs may be suggestive of aberrant clonal expansions, such expansions are not unusual for classical GCs. 14,28,30,35 To address the question whether PTGCs might represent preneoplastic lesions, we analyzed in each of 2 patients the Ki67 ϩ cells from 2 different PTGCs.…”

Molecular Single-Cell Analysis of Hodgkin- and Reed-Sternberg Cells Harboring Unmutated Immunoglobulin Variable Region Genes

“…Briefly, rearranged V H -, V -, and V -genes were amplified using family-specific framework region I (FRI) V-gene primers and two sets of J H -, J -, and J -primers in a semi-nested approach (Bräuninger et al, 1999b;Kanzler et al, 1996a). In some experiments, IgH-and Ig -gene rearrangements were amplified using V H leader- (Bräuninger et al, 1997) and V leader-specific primers instead of V H -and V -FRI-primers. The following V leader family-specific primers were used: 5Ј-GCTCAGCTCCTGGGGCTCCTGC-3Ј for V 1L, 5Ј-CTCAGCTYCTGGGGCTGCTAATGCT-3Ј for V 2L, 5Ј-CTCAGTTAGGACCCAGABGGAACCAT-3Ј for V 3L, and 5Ј-GGGGCAGCAAGATGGTGTTGCAGA-3Ј for V 4L.…”

Validation of immunoglobulin gene rearrangement detection by PCR using commercially available BIOMED-2 primers

“…Further support for this concept comes from the surprising result of moderate clinical sensitivity (35%) using the primer sets to evaluate Hodgkin lymphoma. Although the neoplastic cells in both nodular lymphocyte predominant and classical Hodgkin lymphoma contain rearranged IG genes, 6,7 they contain extensive somatic hypermutation and represent a Non-specific Results From the IGK-A tube 120-160 bp size range. Representative tracings from a negative sample (a) and a positive sample (b) showing a prominent reactive peak in the negative tracing (arrow) that might mistakenly be interpreted as positive.…”