466. Artesunate Inhibits the Immortalization of Human Umbilical Cord Blood Lymphocytes (HUCL) by Epstein-Barr virus (EBV)
Session: Poster Abstract Session: Prevention and Treatment of Viral Infections
Thursday, October 3, 2013
Room: The Moscone Center: Poster Hall C
Posters
  • PneumoPoster_BZK_9-26-13.pdf (3.6 MB)
  • Background:  Concentrations of artesunate that have been found to inhibit lytic Epstein-Barr virus (EBV) replication are in the range of 5-10 µM [Efferth, Clin Infect Dis 2008;47:804; Auerochs, J. Virol Meth 2011; 173:334].  We tested whether artesunate can inhibit EBV immortalization of human umbilical cord blood lymphocytes (HUCL).    

    Methods: Artesunate (Sigma-Aldrich, St Louis, MO) is dissolved in DMSO.  Cord blood was Ficoll-hypaque separated, counted and resuspended at 2 x 106 / ml in RPMI with 10% fetal calf serum using standard methodologies [Katz, J Infect Dis 1988; 157:299].  The source of immortalizing EBV was standardized, filtered aliquots of supernatant from B95-8 cells.  Plates were prepared in duplicate.  Eight replicates of each dilution of drug, virus or control were plated.  Every well of each plate was seeded with 2 x 105cord blood cells.  Every plate contained the following negative and positive immortalization control wells, respectively: HUCL and medium, HUCL and B95-8 supernatant.  Other rows contained B95-8 supernatant, cord blood cells and either 5 or 10 µM of DMSO (another positive immortalization control) or B95-8 supernatant, cord blood cells, and either 5 or 10 µM of artesunate.  Plates were examined weekly for immortalization for at least 2 months and fed with fresh medium.  Immortalization was scored visually and by outgrowth of immortalized cell lines [Katz, J Infect Dis 1988; 157:299].

    Results: The results of 4 different experiments are pooled.  As expected, no immortalization was seen in the negative control wells (0/29), while 11/31 wells were immortalized when B95-8 supernatant was added.  When 5 µM of DMSO was added to the B95-8 supernatant, 5/29 wells demonstrated immortalization; when 10 µM of DMSO was added to the B95-8 supernatant, 3/15 wells demonstrated immortalization.  None of 29 wells demonstrated immortalization when 5 µM of artesunate was added to the B95-8 supernatant, and none of 16 wells demonstrated immortalization when 10 µM of artesunate was added to the B95-8 supernatant. 

    Conclusion: Five or ten µM of artesunate can inhibit immortalization of HUCL by EBV. This drug should be further tested as a potential agent against EBV infection.

    Ben Katz, MD, Pediatrics, Ann and Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL; Northwestern University Feinberg School of Medicine, Chicago, IL, Joseph Quintas, Infectious Diseases, Ann and Robert H Lurie Children's Research Center of Chicago, Chicago, IL and William Kabat, BS, Infectious Diseases, Ann and Robert H Lurie Children's Reseacr Center of Chicago, Chicago, IL

    Disclosures:

    B. Katz, None

    J. Quintas, None

    W. Kabat, None

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