Activated macrophages and antibodies against the plant lectin, GSI-B4, recognize the same tumor-associated structure (TAS).
Activated macrophages that were stabilized with either formalin or glutaraldehyde absorbed two polypeptides (Mr 100,000 and 60,000) from detergent extracts of all of the tumor cell lines tested, but not from detergent extracts of normal human peripheral blood lymphocytes. A major polypeptide (Mr 95,000) was retained from spent culture media of tumor cell lines. Polypeptides with molecular sizes of 100,000 and 60,000 daltons were also adsorbed by activated macrophages from detergent extracts of chicken embryo cell membranes, suggesting an oncofetal nature for these proteins. The 100,000 dalton polypeptide, but not the 60,000 dalton component, was found to be available to lactoperoxidase-catalyzed cell surface iodination. Polypeptides with identical molecular sizes could be adsorbed to immobilized galactopyranoside, indicating that they are vertebrate lectins. Activated macrophages and affinity adsorbents prepared by the covalent coupling of galactopyranoside to agarose also bind the plant lectin isolectin B4 prepared from the seeds of Griffonia simplicifolia. On the basis of these findings, we put forth the hypothesis that macromolecules of the same specificity, that is affinity to galactopyranosyl residues, must show homologies in their binding sites. We have predicted therefore that antisera prepared against this plant lectin should cross-react with galactopyranosyl-binding vertebrate lectins present on the surface of tumor cells. In this communication, we also report the generation of hybridomas that produce antibodies reactive with both the plant and vertebrate lectins. Inhibition experiments that make use of various mono- and disaccharides suggest that the specificities of these antibodies are for determinants intimately associated with the galactosyl binding site on the lectin molecule. Two of the antibodies were found to have moderate selectivity for tumor cells when tested in an immunohistochemical procedure that made use of fresh-frozen or paraffin-embedded sections of human biopsy material. These two antibodies on immunoblots of tumor cell membrane extracts reacted with a polypeptide with an apparent molecular size of 100,000 daltons.
D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology*; D002460:Cell Line; D004622:Embryo, Mammalian / Q000276:immunology; D005690:Galactose / Q000276:immunology; D006168:Guinea Pigs; D006801:Humans; D007157:Immunologic Surveillance; D037102:Lectins / Q000276:immunology*; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D037121:Plant Lectins
D000951:Antigens, Neoplasm; C045475:Griffonia simplicifolia lectins; D037102:Lectins; D037121:Plant Lectins; D005690:Galactose