DETECTION OF IMMUNOGLOBULIN HEAVY-CHAIN GENE REARRANGEMENT AT THE SINGLE-CELL LEVEL IN MALIGNANT-LYMPHOMAS - NO REARRANGEMENT IS FOUND IN HODGKIN AND REED-STERNBERG CELLS

Hodgkin and Reed-Sternberg (HRS) cells, the neoplastic cells of Hodgki n's disease (HD) represent only a minority of the cellular infiltrate in affected tissue. Therefore, rearrangements of the immunoglobulin he avy-chain (IgH) gene detected in DNA extracted from an entire Hodgkin' s lymph node cannot be attributed to the HRS cells and cannot be used as an argument for the B-cell origin of HRS cells. We developed a new method for the amplification of rearranged DNA of the IgH gene from si ngle cells. Using 6 ''forward primers'' which were constructed corresp onding to consensus sequences of the 6 known families of the IgH varia ble (V) region (framework region I) and a mix of 2 ''reverse primers'' corresponding to consensus sequences of the different joining (J) seg ments, rearrangements of all 6 V-families were detected in human perip heral blood lymphocytes. Rearranged IgH DNA could be amplified from si ngle cells of B-cell lymphoma-cell lines and from 13 patients with B-c ell non-Hodgkin's lymphomas. However, analysis of HRS cells isolated f rom lymph nodes of 13 patients with Hodgkin's disease did not show any rearrangement of the IgH gene locus. These findings, obtained by poly merase chain reaction (PCR) on isolated single HRS cells, contrast wit h previous studies that used Southern-blot analysis of entire tissues affected by Hodgkin's disease. We conclude that the neoplastic HRS cel ls in Hodgkin's disease-with the possible exception of the nodular par agranuloma subtype-are probably not derived from B cells. (C) 1994 Wil ey-Liss, Inc.