Single cell PCR for the analysis of Hodgkin's disease: Four years later

Abstract

Background: Single cell-based studies represent a promising alternative to conventional molecular approaches in the study of Hodgkin's disease since the malignant Hodgkin and Reed-Sternberg cells (H and RS) represent only a small minority of the cellular infiltrate in affected nodes. Methods: Single cell polymerase chain reaction (PCR) assays were developed for the analysis of specific genomic DNA sequences and the detection of gene expression. Single H and RS cells were isolated by micromanipulation from cytospin slides or fresh cell suspensions after staining with an anti-CD 30 MoAB. Results: The status of oncogenes and immune receptor genes was examined by DNA-PCR. So far, no IgH or TCR gamma rearrangements were detected in H and RS cells of T- and B-antigen negative classical Hodgkin's cases but were detected in two cases of nodular paragranuloma. Global cDNA amplification was successfully performed from single H and RS cells, and specific gene transcripts were detected with a novel PCR method. Conclusion: Single cell PCR is a novel and promising method that will help to elucidate many of the open questions in the biology of Hodgkin's disease. In the case of contradictory results, collaborations between different groups utilizing similar approaches have to be performed.