SINGLE-CELL PCR FOR THE ANALYSIS OF HODGKINS-DISEASE - 4 YEARS LATER

Background: Single cell-based studies represent a promising alternativ e to conventional molecular approaches in the study of Hodgkin's disea se since the malignant Hodgkin and Reed-Sternberg cells (H & RS) repre sent only a small minority of the cellular infiltrate in affected node s. Methods: Single cell polymerase chain reaction (PCR) assays were de veloped for the analysis of specific genomic DNA sequences and the det ection of gene expression. Single H & RS cells were isolated by microm anipulation from cytospin slides or fresh cell suspensions after stain ing with an anti-CD 30 MoAB. Results: The status of oncogenes and immu ne receptor genes was examined by DNA-PCR. So far, no IgH or TCR B-ant igen negative classical Hodgkin's cases but were detected in two cases of nodular paragranuloma. Global cDNA amplification was successfully performed from single H & RS cells, and specific gene transcripts were detected with a novel PCR method. Conclusion: Single cell PCR is a no vel and promising method that will help to elucidate many of the open questions in the biology of Hodgkin's disease. In the case of contradi ctory results, collaborations between different groups utilizing simil ar approaches have to be performed.