Differential binding of soybean agglutinin to human neuroblastoma cell lines: potential application to autologous bone marrow transplantation.
Normal human bone marrow cells were mixed with neuroblastoma cells from four different human cell lines, and the cell mixtures were separated by differential agglutination with soybean agglutinin (SBA). The unagglutinated cell fraction, previously shown to be highly enriched for the hematopoietic pluripotential stem cells and capable of reconstituting lethally irradiated adult patients with acute leukemia, was further fractionated by affinity chromatography on the lectin conjugated to Sepharose 6MB beads. Two independent assays, one using radiolabeling of the tumor cells and the other based on cloning of the neuroblastoma cells on agar, showed that the agglutination step alone removes 64-76% of the radiolabeled neuroblastoma cells and 85-98% of the clonogenic cells from the tumor/bone marrow cell mixture. Passage of the unagglutinated radiolabeled cells through SBA-Sepharose columns results in further purging of 28-53% of the neuroblastoma cells. Thus a combination of the two methods affords only one-log depletion for the neuroblastoma cells, compared to a three-log depletion achieved for a T-cell leukemia line CEM tested in parallel. It seems therefore that the agglutination technique, or the use of SBA-Sepharose columns, can be used only as a preliminary step for the purging of neuroblastoma cells from involved human bone marrow preparations. Staining with fluorescein isothiocyanate-conjugated SBA of nine different neuroblastoma cell lines, including the four tested in the fractionation studies, showed that more than 98% of the cells, of all the cell lines tested, specifically bind to the lectin, whereas no specific binding can be detected on the stem cell-enriched bone marrow cell fraction. However, the total number of receptors on the neuroblastoma cells is small compared to that of line CEM or normal granulocytes, which are strongly agglutinated by SBA. It seems therefore that the quantitative difference in the total number of SBA receptors is a crucial factor for purging by the agglutination technique or by affinity chromatography. Although these results show limitations to the use of both methods, this study establishes that all neuroblastoma cell lines tested express receptors for the lectin. Improved purging of neuroblastoma cells may possibly be achieved by targeting SBA-bound toxins or magnetic spheres to these receptors.
Y
Y
Reisner
J
J
Gan
D000371:Agglutination; D016026:Bone Marrow Transplantation; D002460:Cell Line; D002847:Chromatography, Agarose; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006098:Granulocytes / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D007938:Leukemia / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism*; D037121:Plant Lectins; D030262:Soybean Proteins; D013194:Staining and Labeling; D013861:Thiocyanates; D014182:Transplantation, Autologous; D010835:Phytohemagglutinins
D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D037102:Lectins; D037121:Plant Lectins; D030262:Soybean Proteins; D013861:Thiocyanates; C028824:soybean lectin; D010835:Phytohemagglutinins