X-chromosome inactivation analysis of isolated Reed-Sternberg cells in nodular sclerosing Hodgkin's disease

In spite of several studies of immunoglobulin (Ig) gene rearrangements in whole Hodgkin's disease (HD) tissues and in isolated single Reed-Sternberg (RS) cells, the issue of clonality of the RS cell in HD remains incompletely resolved. Analysis of X-chromosome inactivation patterns (XCIPs) can be used to determine whether cell populations are clonal in origin, By FCR amplification of the human androgen receptor (HUMARA) loci using nested primers, we have studied XCIPs in six cases of HU of the nodular sclerosing (NS) subtype in which individual RS cells were isolated by micromanipulation from formalin-fixed, paraffin-embedded tissue sections immunostained with CD30 or CD15, In order to assess whether a clonal population of RS cells might be present in NSHD tissues, we compared the XCIPs obtained from whole NSHD tissues with those obtained from single RS cells harvested from the same tissues. Whole tissues from all six cases of NSHD showed balanced HUMARA allelic patterns, whereas an average of 83% (range 77-91%) of single RS cells from each of the six tissues expressed the same high- or low-molecular weight allele, suggesting that a clonal population of RS cells was likely to be present in each case. These data are consistent with the presence of a clonal population of RS cells in NSHD tissues.