DETECTION OF CLONAL HODGKIN AND REED-STERNBERG CELLS WITH IDENTICAL SOMATICALLY MUTATED AND REARRANGED V-H GENES IN DIFFERENT BIOPSIES IN RELAPSED HODGKINS-DISEASE

Hodgkin's disease (HD) represents a malignant lymphoma in which the pu tative malignant Hodgkin and Reed-Sternberg (H-RS) cells are rare and surrounded by abundant reactive cells. Single-cell analyses showed tha t H-RS cells regularly bear clonal Ig gene rearrangements. However, th ere is little information on the clinical evolution of HD in a given p atient. In this study, we used the single-cell polymerase chain reacti on (PCR) to identify H-RS cells with clonal Ig gene rearrangements in biopsy specimens of patients with relapsed HD. The obtained clonal var iable region heavy-chain (VH) gene rearrangements were used to constru ct tumor-clone-specific oligonucleotides spanning the complementarity determining region (CDR) III and somatically mutated areas in the rear ranged VH gene. A number of biopsies were obtained during a period of 3 years from two HD patients. H-RS cells with identical VH rearrangeme nts were detected in two separate infiltrated lymph nodes from one pat ient with nodular sclerosis HD. In a second patient with mixed cellula rity HD subtype, clonal VH rearrangements with identical sequences wer e detected in infiltrated spleen and two lymph node biopsies. Despite the high sensitivity of the PCR method used tone clonal cell in 10(5) mononuclear cells), residual H-RS cells were not found in peripheral b lood, leukapheresis material, purified CD34(+) stem cells or bone marr ow, The results show that different specimens from relapsed patients s uffering from classical HD carry the same clonotypic IgH rearrangement s with identical somatic mutations, demonstrating the persistence and the dissemination of a clonal tumor cell population. Thus, PCR assays with CDRIII-specific probes derived from clonal H-RS cells are of clin ical importance in monitoring the dissemination of HD and tumor progre ssion and could be useful for analysis of minimal residual disease aft er autologous stem cell transplantation. (C) 1998 by The American Soci ety of Hematology.