title: CD19 selection improves the sensitivity of B cell lymphoma detection.
abstract: Reinfusion of residual tumor cells into B cell non-Hodgkin's lymphoma (B-NHL) patients during autologous transplantation may be an important cause of disease relapse. Determining the extent to which B-NHL cells are present in autologous progenitor cell products and if the presence of residual B-NHL cells is predictive of relapse will require extremely sensitive methods of detecting rare B-NHL cells. We attempted to improve the sensitivity of polymerase chain reaction (PCR)-based detection of rare B-NHL cells by preselecting CD19+ cells using an immunomagnetic column. To measure detection sensitivity, we prepared samples containing different levels of B-NHL cell contamination by mixing B-NHL cell lines containing the chromosomal translocation t(14;18) bcl-2/JH) with control leukapheresis samples. DNA extracted from each CD19-selected sample and from each matched nonselected sample was added to a PCR to amplify the bcl-2/JH breakdown junction. CD19 preselection improved the sensitivity of detection of t(14;18)-positive B-NHL cells 115-fold, so that B-NHL cells at a concentration of 1 tumor cell per 1 x 10(6) hematopoietic cells were detected in every specimen evaluated. t(14;18)-positive cells were not detected in any of 13 control leukapheresis specimens. We conclude that a combination of CD19 preselection and PCR amplification may improve the sensitivity of detection of rare lymphoma cells by two orders of magnitude without a significant decrease in specificity.
authors.affiliation: Department of Pathology, University of Colorado Health Sciences Center, Denver 80262, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Pugh
authors.forename: M A
authors.initials: MA
authors.lastname: Bitter
authors.forename: E J
authors.initials: EJ
authors.lastname: Shpall
authors.forename: L S
authors.initials: LS
authors.lastname: Hami
authors.forename: D M
authors.initials: DM
authors.lastname: Wolf
authors.forename: W A
authors.initials: WA
authors.lastname: Franklin
mesh_terms: D018941:Antigens, CD19 / Q000276:immunology; D004247:DNA / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D006801:Humans; D007937:Leukapheresis; D016393:Lymphoma, B-Cell / Q000097:blood* / Q000175:diagnosis; D018365:Neoplasm, Residual / Q000097:blood* / Q000175:diagnosis; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D018941:Antigens, CD19; D004247:DNA