Relationships between degree of binding, cytotoxicity and cytoagglutinating activity of plant-derived agglutinins in normal lymphocytes and cultured leukemic cell lines.
OBJECTIVE
To clarify the relationships between the degree of lectin-cell binding, cytotoxicity and cytoagglutinating activity of plant-derived lectins in normal lymphocytes and cultured leukemic cell lines.


METHODS
Plant lectins with different quaternary structures and saccharide specificity were used: Dolichos biflorus agglutinin (DBA), Soybean agglutinin (SBA) and Wheat germ agglutinin (WGA). The leukemic cell lines used were: Jurkat, MOLT-4, RPMI-8402, HPB-ALL, CCR-HSB-2 and BALL-1 (derived from acute lymphoblastic leukemia); Raji and Daudi (derived from Burkitt's lymphoma); K-562 (derived from myelogenous leukemia). The lectin-cell binding was detected microscopically and fluorimetrically using FITC-conjugated lectins. Cytotoxicity was estimated by the CellTiter-Glo luminescent cell viability assay, and cytoagglutinating activity by a spectrophotometric method.


RESULTS
The binding of DBA and SBA to normal lymphocytes was negligible, while their binding to leukemic cells increased markedly with increasing lectin concentration. Analogous results were obtained for WGA. However, it was found that WGA also interacted to a significant degree with normal lymphocytes. The degree of lectin-cell binding increased in the order: DBA<SBA<WGA. The cytoagglutinating activity and cytotoxicity of lectins increased in the same order. DBA did not exhibit a cytotoxic effect against normal or leukemic cells, and showed a poor cytoagglutinating activity only in MOLT-4, CCR-HSB-2 and BALL-1 cells. SBA exhibited poor cytotoxicity against Jurkat, RPMI-8402, HPB-ALL and CCR-HSB-2 cells, but a well-defined cytotoxicity against Raji and Daudi cells. SBA showed poor cytoagglutinating activity in leukemic cells. In contrast, WGA at concentrations higher than 0.05 microM showed high cytotoxicity against all leukemic cell lines tested as well as against normal lymphocytes. WGA also showed a well-expressed cytoagglutinating effect in all cell lines except normal lymphocytes. There was a moderate inverse correlation between cell viability and the velocity of cytoagglutination ( r=-0.56, P<0.001), and a good correlation between cell viability and the degree of lectin-cell binding ( r=-0.75, P<0.001). There was a low positive correlation between the velocity of cytoagglutination and the degree of lectin-cell binding ( r=0.43, P<0.001).


CONCLUSIONS
The results suggest that the lectins that bound most strongly to leukemic cells expressed higher cytotoxic and cytoagglutinating activities.
Ohba
Hideki
H
Single-Molecule Bioanalysis Laboratory, National Institute for Advanced Industrial Science and Technology, AIST-Shikoku, 2217-14 Hayashi-cho, Takamatsu, 761-0395, Kagawa, Japan.
Bakalova
Rumiana
R
D000371:Agglutination / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D037102:Lectins; D007938:Leukemia / Q000097:blood / Q000378:metabolism*; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism*; D037121:Plant Lectins / Q000378:metabolism* / Q000633:toxicity*; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D030262:Soybean Proteins / Q000378:metabolism / Q000633:toxicity; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins / Q000378:metabolism / Q000633:toxicity; D010835:Phytohemagglutinins
D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D037102:Lectins; D037121:Plant Lectins; D011956:Receptors, Cell Surface; D030262:Soybean Proteins; D014909:Wheat Germ Agglutinins; C036554:dolichos biflorus agglutinin; C028824:soybean lectin; D010835:Phytohemagglutinins