Antigen-specific primary immune response of human B-lymphocytes after in vitro immunization with GM3 ganglioside¹

Article type: Research Article

Authors: Alfonso, Mauro^{a; e} | Lanne, Boel^b | Ifversen, Peter^c | Vázquez, Ana M.^a | Pérez, Rolando^a | Portoukalian, Jacques^d | Zeuthen, Jasper^{e; *}

Affiliations: [a] Center of Molecular Immunology, Dept. of Research and Development, Havana 11600, Cuba | [b] Dept. of Medical Biochemistry, University of Göteborg S-41390 Göteborg, Sweden | [c] Lund University, Dept. of Immunotechnology, S-22007 Lund, Sweden | [d] INSERM, Centre Léon Bérard, Laboratory of Immunology and Experimental Oncology, F-69373 Lyon Cedex 08, France | [e] Danish Cancer Society, Dept. of Tumor Cell Biology, DK-2100 Copenhagen, Denmark

Correspondence: [*] Correspondence and reprint requests to: Prof. Jesper Zeuthen, Department of Tumor Cell Biology, Division of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, DK-2100 Copenhagen, Denmark.

Note: [1] A version of this paper was presented at the Fourth International Conference on Human Antibodies and Hybridomas, Amsterdam, The Netherlands, 26–28 April 1995.

Abstract: In vitro immunization of human B-lymphocytes was performed with liposomes containing the monosialoganglioside GM3, with or without either complete tetanus toxoid or a synthetic T helper epitope derived from tetanus toxin (determinant 830–843). The immunized B-cells were Epstein–Barr virus transformed and the human anti-ganglioside antibody response was evaluated using an indirect ELISA against different mono- and disialogangliosides. Clones producing antigen-specific human antibodies of the IgM isotype against the ganglioside GM3 used as the immunogen were selected and one clone, IM-11, was further characterized. In addition, a method of positive selection using GM3-coated magnetic beads has been developed which allowed us to rescue unstable clones. The binding of the human antibody M-11 to a large panel of glycosphingolipids separated on thin-layer plates was studied. The human MAb IM-11 was found to bind strongly to NeuAcGM3, IV3NeuAcnLc4 and sulfate containing glycosphingolipids and weakly to NeuGcGM3. Immunohistological staining of melanoma and breast cancer biopsy sections showed a selective reactivity of IM-11 with tumor cells which varied among different tumors.

Keywords: In vitro immunization, human monoclonal antibody, GM3 ganglioside, sulfated glycolipids

DOI: 10.3233/HAB-1995-6304

Journal: Human Antibodies, vol. 6, no. 3, pp. 102-112, 1995