Erythrocyte-Based Targeted Release to Macrophages of an Azidothymidine Homodinucleotide Prevents Retroviral Infection

  • Umberto Benatti
  • Marco Giovine
  • Gianluca Damonte
  • Antonio De Flora
  • Rosamund Williams
  • Sandra Gessani
  • Giorgio Brandi
  • Anna Casabianca
  • Alessandra Fraternale
  • y Mauro Magnani

Summary

Mononuclear phagocytes, representing a known reservoir of HIV-1, feature a poor response to the nucleoside analogues currently used in AIDS therapy. The main reason is a weak phosphorylating activity to start metabolic activation that terminates with the intracellular synthesis of the retroviral reverse transcriptase inhibitors, i.e. the corresponding nucleoside triphosphates. The azidothymidine homodinucleotide (AZTp2AZT) recently designed and synthesized in our laboratories proved to be suitable to overcome this limitation. AZTp2AZT was encapsulated in erythrocytes where it showed a remarkable stability. This allowed to selectively perturb the membrane of the AZTp2AZT-loaded erythrocytes, thereby triggering subsequent opsonization and uptake by macrophage cultures. Macrophages exposed to carrier erythrocytes showed a significant resistance to infection by immunodeficiency viruses. Protection was observed with human, feline and murine macrophages against infection by HIV-1, FIV and LP-BM5 retroviral strains, respectively. These data demonstrate that AZTp2AZT-loaded and opsonized erythrocytes represent an efficient “trojan horse” resulting in significant protection of macrophages from retroviral infection.

Keywords

Nucleoside Analogue Mononuclear Phagocyte Macrophage Culture Nodeficiency Virus Diphosphate Ribose 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Benatti, U., Giovine, M, Damonte, G., Gasparini, A., Scarfi, S., De Flora, A., Fraternale, A., Rossi, L., Magnani, M. Azidothymidine homodinucleotide-loaded erythrocytes as bioreactors for slow delivery of the antiretroviral drug azidothymidine. Biochem. Biophys. Res. Comm., 220, 20–25, 1996.PubMedCrossRefGoogle Scholar
  2. 2.
    Fraternale, A., Casabianca, A., Rossi, L., Chiarantini, L., Brandi, G., Aluigi, G., Schiavano, G.F., Magnani, M. Inibition of murine AIDS by combination of AZT and dideoxycytidine 5′-triphosphate. J. Acquir. Immune Defic. Syndr. 12, 164–173, 1996.CrossRefGoogle Scholar
  3. 3.
    Gendelman H.E., Meltzer M.S. Mononuclear phagocytes and the human immunodeficiency virus. Curr. Opin. Immunol. 2, 414–419, 1990.CrossRefGoogle Scholar
  4. 4.
    Giovine, M., Scarfi, S., Gasparini, A., Millo, E., Damonte, G., De Flora, A., Magnani, M., Fraternale, A., Rossi, L., Williams, R., Benatti, U. Synthesis, characterization and erythrocyte encapsulation of an azidothymidine homodinucleotide behaving as a prodrug of azidothymidine. Biological Carriers in Medicine (U. Sprandel and J.L. Way Eds.) Plenum Press, New York, in press.Google Scholar
  5. 5.
    Lewis, W., Dalakas, M.C., Mitochondrial toxicity of antiviral drugs. Nat. Med. 1, 417–422, 1995.PubMedCrossRefGoogle Scholar
  6. 6.
    Magnani, M., Rossi, L., Brandi, G., Schiavano, G.F., Montroni, M., Piedimonte, G. Targeting antiretroviral nucleoside analogues in phosphorylated form to macrophages: in vitro ed in vivo studies. Proc. Natl. Acad. Sci. USA 89, 6477–6481, 1992.PubMedCrossRefGoogle Scholar
  7. 7.
    Magnani, M., Giovine, M., Fraternale, A., Damonte, G., Rossi, L., Scarfi, S., Benatti, U., De Flora, A. Red Blood Cells as a Delivery System for AZT. Drug Delivery, 2, 57–61, 1995.CrossRefGoogle Scholar
  8. 8.
    Magnani, M., Casabianca, A., Fraternale, A., Brandi G., Gessani, S., Williams, R., Giovine, M., Damonte, G., De Flora, A., Benatti, U. Synthesis and targeted delivery of an azidothymidine homodinucleotide conferring protection to macrophages against retroviral infection. Proc. Natl. Acad. Sci. USA 93, 4403–1408, 1996.PubMedCrossRefGoogle Scholar
  9. 9.
    Parker, W.B., Cheng, Y.C., Mitochondrial toxicity of antiviral analogs. J. NIH Res. 6, 57–61, 1994.Google Scholar
  10. 10.
    Perno, C.F., Yarchoan, R., Cooney, D.A., Hartman, N.R., Gartner, S., Popovic, M., Hao, Z., Gerrard, T.L., Wilson, Y.A., Johns, D.G. and Broder, S. J. Inhibition of human immunodeficiency virus (HIV-1/HTLV-II-IBa-L) replication in fresh and cultured human peripheral blood monocytes/macrophages by azidothymidine and related 2′, 3′-′dideoxynucleosides. J. Exp. Med. 168, 1111–1125, 1988.PubMedCrossRefGoogle Scholar
  11. 11.
    Perno CF., Santoro N., Balestra, E., Aquaro S., Lazzarino G., Di Pierro D., Tavazzi B., Balzarini, J., Garaci, E., Grimaldi, S. and Caliò, R. Red blood cells mediated delivery of 9-(2-phosphonyl-methoxyethyl) adenine to primary macrophages: efficiency, metabolism and activity against human immunodeficiency virus or herpes simplex virus. Antiv. Res. 33, 153–164, 1997.CrossRefGoogle Scholar
  12. 12.
    Robbins, B.L., Rodman, J., McDonald, C, Srinivas, R.V., Flynn, P.M., Fridland, A. Enzymatic assay for measurement of zidovudine triphosphate in peripheral blood mononuclear cells. Antimicrob. Agents Chemother. 38, 115–121, 1994.PubMedCrossRefGoogle Scholar
  13. 13.
    Rossi, L., Brandi, G:, Fraternale, A., Schiavano, G.F., Chiarantini, L., Magnani, M. Inibition of murine retrovirus induced immunodeficiency disease by dideoxycytidine and dideoxycytidine 5′ triphosphate J. Acquir. Immune Defic. Syndr. 6, 1179–1186, 1993.Google Scholar
  14. 14.
    Zocchi, E., Guida, L., Franco, L., Silvestro, L., Guerrini, M., Benatti, U. and De Flora, A. Free Adenosine diphosphate ribose in human erythrocytes: pathways of intraerythrocytic conversion and non-enzymic binding to membrane proteins. Biochem. J. 295, 121–130, 1993.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1997

Authors and Affiliations

  • Umberto Benatti
    • 1
  • Marco Giovine
    • 1
  • Gianluca Damonte
    • 1
  • Antonio De Flora
    • 1
  • Rosamund Williams
    • 2
  • Sandra Gessani
    • 3
  • Giorgio Brandi
    • 4
  • Anna Casabianca
    • 5
  • Alessandra Fraternale
    • 5
  • y Mauro Magnani
    • 5
  1. 1.Institute of BiochemistryUniversity of GenoaGenoaItaly
  2. 2.Lepetit Research CenterGerenzano, VareseItaly
  3. 3.Laboratory of VirologyIstituto Superiore di SanitàRomeItaly
  4. 4.Institute of HygieneUniversity of UrbinoUrbinoItaly
  5. 5.Institute of Biochemistry “Giorgio Fornaini”University of UrbinoUrbinoItaly

Personalised recommendations