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Lymphocyte predominance Hodgkin's disease: lineage and clonality
determination using a single-cell assay
J Delabie, A Tierens, G Wu, DD Weisenburger and WC Chan
Department of Pathology and Microbiology, University of Nebraska Medical
Center, Omaha 68198-3135.
Lymphocyte predominance Hodgkin's disease (LPHD) is a clinically indolent
condition. Although there is evidence that the putative neoplastic cell in
this disease, the "L&H" cell, is of B-cell lineage, there is
conflicting data concerning the clonality of these cells. Our study was
aimed at clarifying the issue of lineage and clonality of the L&H cells
of LPHD using a single-cell assay. Four cases of LPHD were studied. To
circumvent the difficulties of obtaining fresh tissue and to be able to
study representative cases, a new method was developed to obtain
single-cell suspensions of L&H cells from archival formalin- fixed
paraffin-embedded tissue. Single L&H cells were identified by
morphology and immunostaining for epithelial membrane antigen, isolated
using a micropipette, and subjected to polymerase chain reaction (PCR)
amplification of the complematarity determining region 3 (CDR3) of the Ig
heavy chain (IgH) gene, which is B-cell clone-specific. The PCR products
were size-fractionated by polyacrylamide gel electrophoresis and
representative products were directly sequenced. Single T cells and small B
cells were also isolated from the tissues and used as negative and positive
controls, respectively. In all four cases of LPHD, the IgH CDR3 of single
L&H cells could be amplified. Within each case, the IgH CDR3 of single
L&H cells was found to be of different length or of different sequence.
Therefore, our results provide strong evidence for the B-cell origin of the
L&H cells and the polyclonal nature of LPHD.
Volume 84,
Issue 10,
pp. 3291-3298,
11/15/1994
Copyright © 1994 by The American Society of Hematology
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