A rapid two-step method for elimination of bcl-2/IgH positive non-Hodgkin's lymphoma cells from human blood or marrow stem cells, employing immunomagnetic purging with streptavidin-coated ferrofluids.

Abstract

BACKGROUND:

Follicular lymphoma cells may contaminate blood or BM stem cell collections used for autologous transplants and contribute to relapse. We report on the development of a B-cell lymphoma-purging technique, using streptavidin-coated ferrofluids (SAFF) and a high-gradient magnetic-separation device (HGMSD).

METHODS:

Blood or BM samples were spiked with bcl-2 positive lymphoma cells, labeled with biotinylated B cell MAb (CD19, CD20, CD37) then purged by positive selection of CD34(+) cells, followed by negative selection with SAFF and a high HGMSD. Purging efficiency was judged by dual-labeled flow cytometry and by PCR analysis of bcl-2.

RESULTS:

A 6 log purge of bcl-2 lymphoma cells could be achieved by first enriching for CD34(+) cells, followed by negative selection with SAFF and HGMSD. The use of SAFF with HGMSD alone could only accomplish a 2-3 log purge.

DISCUSSION:

CD34 selection from blood or BM of follicular lymphoma patients, followed by negative selection of MAb-labeled cells with SAFF could achieve a 6 log purge. However, cell yields were low (10%) and purging efficacy was limited to low starting concentration of lymphoma cells (< or = 1.0%). This technique, however, may be useful for full-scale clinical purging of minimal residual disease following debulking with a cyclophosphamide or other agents.

• Authors: K F Mangan, J Leonardo, M T Mullaney, L W Terstappen, C Rao, P Liberti
• Journal: Cytotherapy (Cytotherapy) Vol. 1 Issue 4 Pg. 287-93 ( 1999) ISSN: 1465-3249 [Print] England
• PMID: 20426554 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, CD34
• Proto-Oncogene Proteins c-bcl-2
• Streptavidin

Topics

• Antigens, CD34 (biosynthesis)
• B-Lymphocytes (cytology)
• Biotinylation
• Blood Component Removal
• Cell Separation (methods)
• Cell Transplantation
• Flow Cytometry (methods)
• Genes, Immunoglobulin Heavy Chain
• Humans
• Immunomagnetic Separation (methods)
• Lymphoma, Non-Hodgkin (blood, metabolism)
• Magnetics
• Polymerase Chain Reaction (methods)
• Proto-Oncogene Proteins c-bcl-2 (biosynthesis)
• Streptavidin (chemistry)