Classical Hodgkin and Reed-Sternberg cells demonstrate a non-clonal immature B lymphoid lineage: evidence from a single cell assay and in situ hybridization.
Hodgkin and Reed-Sternberg (H & RS) cells are generally considered to be the neoplastic cells of Hodgkin's disease (HD), however such cells are only found in a minority of the lesions. Recently in a few studies on HD, the clonality of H & RS cells was examined, using a single-cell polymerase chain reaction (PCR) examination. To clarify the lineage and clonality of H & RS cells, we performed single cell PCR and in situ hybridization (ISH), and nine cases of classical HD were thus studied. By ISH, the immunoglobulin J chain, and the kappa and lambda light chain were rarely expressed in the H & RS cells, however, no T-cell markers could be detected. The expression of the recombination activating genes (RAG-1, 2) could be determined in the H & RS cells. We isolated CD30+ H & RS cells, CD3 + T cells and CD20 + B cells from suspended materials using a mechanical sorter. We performed single cell PCR in a sorted individual cell, to amplify the complementarity determining region of the Ig heavy chain (IgH) gene and T-cell receptor gamma chain (TCR gamma) gene. In all cases, TCR gamma could be frequently amplified in the T cells, but was only rarely amplified in the H & RS and B cells. In contrast, the IgH was frequently amplified in the H & RS and B cells, but not in the T cells. In addition, the PCR production of the H & RS cells all showed different lengths. The results therefore support the polyclonal nature and immature B lymphoid cell origin of H & RS cells.
Ohshima
K
K
Department of Pathology, School of Medicine, Fukuoka University, Japan.
Suzumiya
J
J
Mukai
Y
Y
Tashiro
K
K
Shibata
T
T
Tanaka
T
T
Kato
A
A
Kikuchi
M
M
D016175:B-Lymphocyte Subsets / Q000473:pathology*; D019070:Cell Lineage; D015322:Gene Rearrangement, B-Lymphocyte; D015329:Gene Rearrangement, T-Lymphocyte; D005803:Genes, Immunoglobulin; D006566:Herpesviridae Infections / Q000473:pathology / Q000821:virology; D004854:Herpesvirus 4, Human / Q000302:isolation & purification; D006689:Hodgkin Disease / Q000473:pathology* / Q000821:virology; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D011946:Receptors, Antigen / Q000235:genetics; D016539:Reed-Sternberg Cells / Q000473:pathology* / Q000821:virology; D016176:T-Lymphocyte Subsets / Q000473:pathology; D014412:Tumor Virus Infections / Q000473:pathology / Q000821:virology
D007143:Immunoglobulin Heavy Chains; D011946:Receptors, Antigen