Antibody-Mediated Targeting of Liposomes

  • John N. Weinstein
  • Lee D. Leserman
  • Pierre A. Henkart
  • Robert Blumenthal
Part of the NATO Advanced Study Institutes Series book series (NSSA, volume 47)


We have studied a number of permutations on the use of antibody and antigen for targeting liposomes in vitro. Our first strategy was to make liposomes with lipid bearing the dinitrophenyl (DNP) hapten. These liposomes bound specifically to cells in three experimental configurations: (i) with TNP-modified human peripheral blood lymphocytes as targets and sheep IgG anti-TNP as cross-linking agent; (ii) with murine myeloma MOPC 315 cells (which bear an IgA with high affinity for nitrophenyl haptens) as target, using endogeneous surface immunoglobulin on the cells as a point of attachment; (iii) with Fc-receptor bearing cells as targets and rabbit anti-TNP as an opsonizing agent. We monitored the interactions by encapsulating carboxyfluorescein and/or methotrexate in the liposomes, and in some experiments by incorporating 14C-dipalmitoyl phosphatidylcholine or a fluorescent marker in the lipid. In each study large numbers of liposomes could be bound specifically to the target cells, but they were internalized in significant numbers only in (iii) when cells capable of Fc-mediated endocytosis were used. The most striking internalization was found with the mouse macrophage line P388D1. Encapsulated methotrexate had a several-fold greater effect on the metabolism of P388D1 (as assayed by 3H-deoxyuridine uptake) than did an equivalent amount of methotrexate free in solution. This finding indicated that an appropriately chosen drug can escape the phagosomal system to exert its effect in the cytoplasm.

Our second principal effort has been to couple immunoglobulin to liposomal phosphatidylethanolamine covalently, or else through covalently coupled Staphylococcus aureus protein A. Coupling is achieved with the heterobifunctional agent N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). This method of coupling results in minimal aggregation and little leakage of vesicle contents. Liposomes bearing covalently coupled monoclonal antibody bind with high specificity to cells with the corresponding determinants.


P388 Cell RAJI Cell Vesicle Content Covalent Coupling Murine Tumor Cell 
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  1. Barbet, J., Machy,P., and Leserman, L.D., 1981, Monoclonal antibody covalently coupled to liposomes: Specific targeting to cells, J. Supramolec. Struct. Cell Biochem., in press.Google Scholar
  2. Cabantchik, Z.I., Volsky, D.J., Ginsberg, H., and Loyter, A., Reconstitution of the erythrocyte anion transport system: in vitro and in vivo approaches, Ann. N.Y. Acad. Sci., U.S.A. 341: 444.Google Scholar
  3. Eisen, H.N., Simms, E.S., and Potter, M., 1968, Mouse myeloma proteins with anti-hapten antibody activity. The protein produced by plasma cell tumor MOPC-315, Biochemistry 7: 4126.PubMedCrossRefGoogle Scholar
  4. Fraley, R., and D. Papahadjopoulos, D., 1981, New generation liposomes: the engineering of an efficient vehicle for intracellular delivery of nucleic acids, T.I.B.S. 9: 467.Google Scholar
  5. Godfrey, W., Doe, B., Wallace, E.F., Bredt, B., and Wofsy, L., 1981, Affinity targeting of membrane vesicles to cell surfaces, Exptl.Cell Research, in press.Google Scholar
  6. Gregoriadis, G., and Neerunjun, D.E., 1975, Homing of liposomes to target cells, Biochem. Biophys. Res. Commun. 65: 537.Google Scholar
  7. Hagins, W.A., and Yoshikami, S., 1978, In: “Vertebrate Photoreceptors,” Fatt, P., and Barlow, H.B., eds., Academic Press, N.Y.Google Scholar
  8. Heath, T.D., Macher, B.A., and Papahadjopoulos, D., 1981, Covalent attachment of immunoglobulins to liposomes via glycosphingolipids, Biochem. Biophys. Acta, 640: 66.CrossRefGoogle Scholar
  9. Huang, L., and Kennel, S.J., 1979, Binding of immunoglobulin G to phospholipid vesicles by sonication, Biochemistry, 18: 1702.PubMedCrossRefGoogle Scholar
  10. Huang, A., Huang., L., and Kennel, S.J., 1980, Monoclonal antibody covalently coupled with fatty acid, J. Biol. Chem., 255: 8015.PubMedGoogle Scholar
  11. Leserman, L.D., Weinstein, J.N., Blumenthal, R., Sharrow, S.O., and Terry, W.D., 1979, Binding of antigen-bearing fluorescent liposomes to the murine myeloma tumor MOPC 315, J. Immunol. 122: 585.PubMedGoogle Scholar
  12. Leserman, L.D., Weinstein, J.N., Moore, J.J., and Terry, W.D., 1980a, Specific interaction of myeloma tumor cells with haptenbearing liposomes containing methotrexate and carboxyfluorescein, Cancer Res., 40: 4768.PubMedGoogle Scholar
  13. Leserman, L.D., Weinstein, J.N., Blumenthal, R., and Terry, W.D., 1980b, Receptor-mediated endocytosis of antibody opsonized liposomes by tumor cells, Proc. Natl. Acad. Sci. U.S.A., 77: 4089.PubMedCrossRefGoogle Scholar
  14. Leserman, L.D., Barbet, J., Kourilsky, F.M., and Weinstein J.N., 1980c, Targeting to cells of fluorescent liposomes covalently coupled with monoclonal antibody or protein A, Nature, 288: 602.PubMedCrossRefGoogle Scholar
  15. Leserman, L.D., and Weinstein, J.N., Receptor mediated binding and endocytosis of drug-containing liposomes by tumor cells, in: “Liposomes and Immunobiology,” Tom, B.H., and Six, H.R., eds., Elsevier, Amsterdam (1980).Google Scholar
  16. Magee, W.E., Cronenberger, J.H., and Thor, D.E., 1978, Marked stimulation of lymphocyte-mediated attack on tumor cells by target-directed liposomes containing immune RNA, Cancer Res., 38: 1173.PubMedGoogle Scholar
  17. Martin, F.D., Hubbell, W.L. and Papahadjopoulos, D., 1981, Immunospecific targeting of liposomes to cells: a novel and efficient method for covalent attachment of Fab’ fragments via disulfide bonds, Biochemistry, 20: 4229.PubMedCrossRefGoogle Scholar
  18. Six, H.R., Uemura, K., and Kinsky, S.C., 1973, Effect of immunoglobulin class and affinity on the initiation of complement-dependent damage to liposomal model membranes sensitized with dinitrophenylated phospholipids, Biochemistry, 12: 4003.PubMedCrossRefGoogle Scholar
  19. Szoka, F., Magnusson, K.E., Wojcieszyn, J., Hou, Y., Derzko, Z. and Jacobson, K., 1981, Use of lectins and polyethylene glycol for fusion of glycolipid-containing liposomes with eukaryotic cells, Proc. Natl. Acad. Sci. U.S.A., 78: 1685.PubMedCrossRefGoogle Scholar
  20. Weinstein, J.N., Yoshikami, S., Henkart, P.A., Blumenthal, R., and Hagins, W.A., 1977, Liposome-cell interaction. transfer and intracellular release of a trapped fluorescent marker, Science, 195: 489.PubMedCrossRefGoogle Scholar
  21. Weinstein, J.N., Blumenthal, R., Sharrow, S.O., and Henkart, P.A., 1978, Antibody-mediated targeting of liposomes; binding to lymphocytes does not ensure incorporation of vesicle contents into the cells. Biochem. Biophys. Acta, 509: 272.PubMedCrossRefGoogle Scholar
  22. Weissmann, G., Bloomgarden, D., Kaplan, R., Cohen, C., Hoffstein, S., Collins, T., Gotleib, A., and Nagle, D., 1975, A general method for the introduction of enzymes, by means of immunoglobulin-coated liposomes, into lysosomes of deficient cells, Proc. Natl. Acad. Sci. U.S.A., 72: 88.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • John N. Weinstein
    • 1
  • Lee D. Leserman
    • 2
  • Pierre A. Henkart
    • 3
  • Robert Blumenthal
    • 1
  1. 1.Section on Membrane Structure and Function, Laboratory of Theoretical Biology, NCINIHBethesdaUSA
  2. 2.Centre d’Immunologie INSERM-CNRS de Marseille-LuminyMarseille Cedex gFrance
  3. 3.Immunology Branch, NCINIHUSA

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