Editorial Type:
Infectious Disease

Antiviral efficacy of nine nucleoside reverse transcriptase inhibitors against feline immunodeficiency virus in feline peripheral blood mononuclear cells

Anita M. Schwartz Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

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Mary Ann McCrackin Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

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 DVM, PhD
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Raymond F. Schinazi Center for AIDS Research, Veterans Affairs Medical Center, Emory University, Atlanta, GA 30322.

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Peter B. Hill Division of Companion Animal Studies, Department of Clinical Veterinary Science, University of Bristol, Langford House, Langford, North Somerset BS40 5DU, England.

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Thomas W. Vahlenkamp Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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Mary B. Tompkins Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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Katrin Hartmann Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

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 Drmedvet, Drhabil
DOI:
https://doi.org/10.2460/ajvr.75.3.273
Volume/Issue:
Volume 75: Issue 3
Online Publication Date:
01 Mar 2014
Restricted access
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Abstract

Objective—To compare cytotoxic effects and antiviral efficacy of 9 nucleoside reverse transcriptase inhibitors (NRTIs) against FIV in feline peripheral blood mononuclear cells.

Sample—Peripheral blood mononuclear cells obtained from 3 specific pathogen–free cats.

Procedures—3 of the 9 NRTIs had not been previously assessed in feline cell lines. Cytotoxic effects were determined by colorimetric quantification of a formazan product resulting from bioreduction of a tetrazolium reagent by viable peripheral blood mononuclear cells; uninfected cells from 1 cat were used in these assays. Cells from all 3 cats were infected with a pathogenic clone of FIV, and in vitro antiviral efficacy of each NRTI was assessed with an FIV p24 antigen capture ELISA.

Results—Cytotoxic effects in feline peripheral blood mononuclear cells were observed only at concentrations > 10 μM for all 9 NRTIs. Comparison of the cytotoxic effect at the highest concentration investigated (500μM) revealed that didanosine and amdoxovir were significantly less toxic than abacavir. All drugs induced a dose-dependent reduction of FIV replication. At the highest concentration investigated (10μM), there was no significant difference in antiviral efficacy among the test compounds.

Conclusions and Clinical Relevance—The evaluated NRTIs had low cytotoxicity against feline peripheral blood mononuclear cells and appeared to be safe options for further in vivo evaluation for the treatment of FIV-infected cats. There was no evidence suggesting that the newly evaluated compounds would be superior to the existing NRTIs for reducing FIV burden of infected cats.

Abstract

Objective—To compare cytotoxic effects and antiviral efficacy of 9 nucleoside reverse transcriptase inhibitors (NRTIs) against FIV in feline peripheral blood mononuclear cells.

Sample—Peripheral blood mononuclear cells obtained from 3 specific pathogen–free cats.

Procedures—3 of the 9 NRTIs had not been previously assessed in feline cell lines. Cytotoxic effects were determined by colorimetric quantification of a formazan product resulting from bioreduction of a tetrazolium reagent by viable peripheral blood mononuclear cells; uninfected cells from 1 cat were used in these assays. Cells from all 3 cats were infected with a pathogenic clone of FIV, and in vitro antiviral efficacy of each NRTI was assessed with an FIV p24 antigen capture ELISA.

Results—Cytotoxic effects in feline peripheral blood mononuclear cells were observed only at concentrations > 10 μM for all 9 NRTIs. Comparison of the cytotoxic effect at the highest concentration investigated (500μM) revealed that didanosine and amdoxovir were significantly less toxic than abacavir. All drugs induced a dose-dependent reduction of FIV replication. At the highest concentration investigated (10μM), there was no significant difference in antiviral efficacy among the test compounds.

Conclusions and Clinical Relevance—The evaluated NRTIs had low cytotoxicity against feline peripheral blood mononuclear cells and appeared to be safe options for further in vivo evaluation for the treatment of FIV-infected cats. There was no evidence suggesting that the newly evaluated compounds would be superior to the existing NRTIs for reducing FIV burden of infected cats.

Contributor Notes

Dr. McCrackin's present addresses are Research Service, Ralph H. Johnson VA Medical Center, 109 Bee St, Charleston, SC 29401, and the Department of Comparative Medicine, Medical University of South Carolina, Charleston, SC 29425.

Dr. Hill's present address is Department of Veterinary Science, School of Animal and Veterinary Sciences, University of Adelaide, Roseworthy, SA 5371, Australia.

Dr. Vahlenkamp's present address is Institute of Virology, Leipzig University, 04103 Leipzig, Germany.

Dr. Hartmann's present address is Clinic of Small Animal Medicine, Ludwig Maximilian University, 80539 Munich, Germany.

This manuscript represents a portion of a thesis submitted by Anita Schwartz to the Faculty of Veterinary Medicine, Ludwig Maximilians University, Munich, Germany, as partial fulfillment of the requirements for a postgraduate doctoral degree.

Supported in part by a Faculty Research Grant from the University of Georgia College of Veterinary Medicine. Dr. Schinazi was supported in part by CFAR NIH grant 2P30-AI-50409 and by the US Department of Veterans Affairs. The reagents didanosine and abacavir were obtained through the NIH AIDS Reagent Program, Division of AIDS, National Institutes of Allergy and Infectious Diseases, National Institute of Health.

Presented in abstract form at the 11th International Feline Retrovirus Research Symposium, Leipzig, Germany, August 2012.

Address correspondence to Dr. Hartmann (hartmann@lmu.de).
Cover American Journal of Veterinary Research
American Journal of Veterinary Research
Publication Date:
01 Mar 2014

  • 1. Pedersen NC, Ho EW, Brown ML, et al. Isolation of a T-lymphotropic virus from domestic cats with an immunodeficiency-like syndrome. Science 1987; 235: 790–793.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 2. Bandecchi P, Matteucci D, Baldinotti F, et al. Prevalence of feline immunodeficiency virus and other retroviral infections in sick cats in Italy. Vet Immunol Immunopathol 1992; 31: 337–345.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 3. Ishida T, Washizu T, Toriyabe K, et al. Feline immunodeficiency virus infection in cats of Japan. J Am Vet Med Assoc 1989; 194: 221–225.

    • Search Google Scholar
    • Export Citation

  • 4. Pedersen NC, Yamamoto JK, Ishida T, et al. Feline immunodeficiency virus infection. Vet Immunol Immunopathol 1989; 21: 111–129.

  • 5. Zhu Y, Antony JM, Martinez JA, et al. Didanosine causes sensory neuropathy in an HIV/AIDS animal model: impaired mitochondrial and neurotrophic factor gene expression. Brain 2007; 130: 2011–2023.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 6. Zhu YQ, Remington KM, North TW. Mutants of feline immunodeficiency virus resistant to 2′,3′-dideoxy-2′,3′-didehydrothymidine. Antimicrob Agents Chemother 1996; 40: 1983–1987.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 7. de Mari K, Maynard L, Sanquer A, et al. Therapeutic effects of recombinant feline interferon-omega on feline leukemia virus (FeLV)-infected and FeLV/feline immunodeficiency virus (FIV)-coinfected symptomatic cats. J Vet Intern Med 2004; 18: 477–482.

    • Search Google Scholar
    • Export Citation

  • 8. Elder JH, Sundstrom M, de Rozieres S, et al. Molecular mechanisms of FIV infection. Vet Immunol Immunopathol 2008; 123: 3–13.

  • 9. Gobert JM, Remington KM, Zhu YQ, et al. Multiple-drug-resistant mutants of feline immunodeficiency virus selected with 2′,3′-dideoxyinosine alone and in combination with 3′-azido-3′-deoxythymidine. Antimicrob Agents Chemother 1994; 38: 861–864.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 10. Hernandez-Santiago BI, Mathew JS, Rapp KL, et al. Antiviral and cellular metabolism interactions between dexelvucitabine and lamivudine. Antimicrob Agents Chemother 2007; 51: 2130–2135.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 11. Palmisano L, Vella S. A brief history of antiretroviral therapy of HIV infection: success and challenges. Ann Ist Super Sanita 2011; 47: 44–48.

    • Search Google Scholar
    • Export Citation

  • 12. Bisset LR, Lutz H, Boni J, et al. Combined effect of zidovudine (ZDV), lamivudine (3TC) and abacavir (ABC) antiretroviral therapy in suppressing in vitro FIV replication. Antiviral Res 2002; 53: 35–45.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 13. Norelli S, El Daker S, D'Ostilio D, et al. Response of feline immunodeficiency virus (FIV) to tipranavir may provide new clues for development of broad-based inhibitors of retroviral proteases acting on drug-resistant HIV-1. Curr HIV Res 2008; 6: 306–317.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 14. Auwerx J, Esnouf R, De Clercq E, et al. Susceptibility of feline immunodeficiency virus/human immunodeficiency virus type 1 reverse transcriptase chimeras to non-nucleoside RT inhibitors. Mol Pharmacol 2004; 65: 244–251.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 15. Remington KM, Chesebro B, Wehrly K, et al. Mutants of feline immunodeficiency virus resistant to 3′-azido-3′-deoxythymidine. J Virol 1991; 65: 308–312.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 16. McCrackin Stevenson MA, McBroom DG. In vitro characterization of FIV-pPPR, a pathogenic molecular clone of feline immunodeficiency virus, and two drug-resistant pol gene mutants. Am J Vet Res 2001; 62: 588–594.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 17. Zhang W, Mauldin JK, Schmiedt CW, et al. Pharmacokinetics of zidovudine in cats. Am J Vet Res 2004; 65: 835–840.

  • 18. Smyth NR, Bennett M, Gaskell RM, et al. Effect of 3′azido-2′,3′-deoxythymidine (AZT) on experimental feline immunodeficiency virus infection in domestic cats. Res Vet Sci 1994; 57: 220–224.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 19. Hartmann K, Donath A, Beer B, et al. Use of two virustatica (AZT, PMEA) in the treatment of FIV and of FeLV seropositive cats with clinical symptoms. Vet Immunol Immunopathol 1992; 35: 167–175.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 20. Levy J, Crawford C, Hartmann K, et al. 2008 American Association of Feline Practitioners' feline retrovirus management guidelines. J Feline Med Surg 2008; 10: 300–316.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 21. Arai M, Earl DD, Yamamoto JK. Is AZT/3TC therapy effective against FIV infection or immunopathogenesis? Vet Immunol Immunopathol 2002; 85: 189–204.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 22. Hartmann K, Donath A, Kraft W. AZT in the treatment of feline immunodeficiency virus infection. Part 2. Feline Pract 1995; 23(6): 13–20.

    • Search Google Scholar
    • Export Citation

  • 23. Smith RA, Remington KM, Preston BD, et al. A novel point mutation at position 156 of reverse transcriptase from feline immunodeficiency virus confers resistance to the combination of (−)-beta-2′,3′-dideoxy-3′-thiacytidine and 3′-azido-3′-deoxythymidine. J Virol 1998; 72: 2335–2340.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 24. Larder BA, Darby G, Richman DD. HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. Science 1989; 243: 1731–1734.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 25. Larder BA, Kemp SD. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science 1989; 246: 1155–1158.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 26. Martins AN, Medeiros SO, Simonetti JP, et al. Phylogenetic and genetic analysis of feline immunodeficiency virus gag, pol, and env genes from domestic cats undergoing nucleoside reverse transcriptase inhibitor treatment or treatment-naive cats in Rio de Janeiro, Brazil. J Virol 2008; 82: 7863–7874.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 27. Phillips TR, Talbott RL, Lamont C, et al. Comparison of two host cell range variants of feline immunodeficiency virus. J Virol 1990; 64: 4605–4613.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 28. Lerner DL, Grant CK, de Parseval A, et al. FIV infection of IL-2-dependent and -independent feline lymphocyte lines: host cells range distinctions and specific cytokine upregulation. Vet Immunol Immunopathol 1998; 65: 277–297.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 29. Joshi A, Vahlenkamp TW, Garg H, et al. Preferential replication of FIV in activated CD4(+)CD25(+)T cells independent of cellular proliferation. Virology 2004; 321: 307–322.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 30. Hubert JJ. Spearman-Karber method. In: Hubert JJ, ed. Bioassay. 2nd ed. Dubuque, Iowa: Hunt Publishing Co, 1984; 65–66.

  • 31. Smyth NR, McCracken C, Gaskell RM, et al. Susceptibility in cell culture of feline immunodeficiency virus to eighteen antiviral agents. J Antimicrob Chemother 1994; 34: 589–594.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 32. Hartmann K, Donath A, Kraft W. AZT in the treatment of feline immunodeficiency virus infection. Part 1. Feline Pract 1995; 23(5): 16–21.

    • Search Google Scholar
    • Export Citation

  • 33. Vahlenkamp TW, De Ronde A, Balzarini J, et al. (R)-9-(2-phosphonylmethoxypropyl)-2,6-diaminopurine is a potent inhibitor of feline immunodeficiency virus infection. Antimicrob Agents Chemother 1995; 39: 746–749.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 34. Balzarini J, Egberink H, Hartmann K, et al. Antiretrovirus specificity and intracellular metabolism of 2′,3′-didehydro-2′,3′-dideoxythymidine (stavudine) and its 5′-monophosphate triesterprodrug So324. Mol Pharmacol 1996; 50: 1207–1213.

    • Search Google Scholar
    • Export Citation

  • 35. Du DL, Volpe DA, Grieshaber CK, et al. In vitro myelotoxicity of 2′,3′-dideoxynucleosides on human hematopoietic progenitor cells. Exp Hematol 1990; 18: 832–836.

    • Search Google Scholar
    • Export Citation

  • 36. Lambert JS, Seidlin M, Reichman RC, et al. 2′,3′-dideoxyinosine (ddI) in patients with the acquired immunodeficiency syndrome or AIDS-related complex. A phase I trial. N Engl J Med 1990; 322: 1333–1340.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 37. White AJ. Mitochondrial toxicity and HIV therapy. Sex Transm Infect 2001; 77: 158–173.

  • 38. van der Meer FJ, Schuurman NM, Balzarini J, et al. Comparative evaluation of the activity of antivirals towards feline immunodeficiency virus in different cell culture systems. Antiviral Res 2007; 76: 198–201.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 39. North TW, North GL, Pedersen NC. Feline immunodeficiency virus, a model for reverse transcriptase-targeted chemotherapy for acquired immune deficiency syndrome. Antimicrob Agents Chemother 1989; 33: 915–919.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 40. Hart S, Nolte I. Long-term treatment of diseased, FIV-seropositive field cats with azidothymidine (AZT). Zentralbl Veterinarmed A 1995; 42: 397–409.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 41. Zhu Y, Vergote D, Pardo C, et al. CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy. FASEB J 2009; 23: 2928–2941.

    • Crossref
    • Search Google Scholar
    • Export Citation

  • 42. Delta Coordinating Committee. Evidence for prolonged clinical benefit from initial combination antiretroviral therapy: Delta extended follow-up. HIV Med 2001; 2: 181–188.

    • Search Google Scholar
    • Export Citation

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