Photodynamic Inactivation of Herpesvirus

  • J. L. Melnick
  • C. Wallis
Part of the Photobiology book series (PB)


Photodynamic inactivation has been known since Raab, at the beginning of the 20th century, observed that acridine was harmless to paramecia in the dark but was lethal when the organisms were exposed to visible light (1). Three decades later viruses were shown to be photosensitive (2, 3). However, assay methods in the 1930s were crude, and no quantitative results were reported. In 1958, Yamamoto (4) reported the first quantitative studies on photodynamic inactivation of bacterial virus; in 1960, Hiatt et al. (5) extended this work to a number of DNA-containing animal viruses, but found that RNA-containing enteroviruses were resistant to photosensitization. We have since learned that naturally photoresistant viruses can be made photosensitive if the virus is grown in cells maintained with medium containing proflavine, neutral red, or acridine orange. During replication of the virus, the photoreactive dye becomes incorporated within the virus structure (6–10). If one looks through the literature, one can see that in most laboratories virus titers could be reduced markedly by “dye-light” treatment, but usually some virus persisted. The point is that, as usually practiced, photoinactivation may not be complete. Transformation by such preparations in which some infectious virus is still present (11) cannot be said to be caused by photoinactivated virus. However, if the procedures described in the papers from our laboratory are carefully followed, photoinactivation can be made total (12–17).


Methylene Blue Herpes Simplex Virus Acridine Orange Live Virus Photodynamic Action 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Raab, O. Z. (1900): Ueber die Wirkung fluorescirender Stoffe auf Infusorien. Z. Biol 39:524–526.Google Scholar
  2. 2.
    Perdrau, J. R., and Todd, C. (1933): Photodynamie action of methylene blue on certain viruses. Proc. R. Soc. Lond. [Biol.] 112:288–298.CrossRefGoogle Scholar
  3. 3.
    Shortt, H. E., and Brooks, A. G. (1934): Photodynamie action of methylene blue on fixed rabies virus. Indian J. Med. Res. 21:581.Google Scholar
  4. 4.
    Yamamoto, N. (1958): Photodynamie inactivation of bacteriophage and its inhibition. J. Bacteriol 75:443–448.PubMedGoogle Scholar
  5. 5.
    Hiatt, C. W., Kaufman, E., Helprin, J. J., et al. (1960): Inactivation of viruses by the photodynamie action of toluidine blue. J. Immunol 84:480–484.PubMedGoogle Scholar
  6. 6.
    Crowther, D., and Melnick, J. L. (1961): The incorporation of neutral red and acridine orange into developing poliovirus particles making them photosensitive. Virology 14:11–21.PubMedCrossRefGoogle Scholar
  7. 7.
    Schaffer, F. L. (1962): Binding of proflavine by photoinactivation of poliovirus propagated in the presence of the dye. Virology 18:412–425.PubMedCrossRefGoogle Scholar
  8. 8.
    Wilson, J. N., and Cooper, P. D. (1962): Photodynamie demonstration of two stages in the growth of poliovirus. Virology 17:195–196.PubMedCrossRefGoogle Scholar
  9. 9.
    Schaffer, F. L., and Hackett, A. J. (1963): Early events in poliovirus-HeLa cell interaction: acridine orange photosensitization and detergent extraction. Virology 21:124–126.PubMedCrossRefGoogle Scholar
  10. 10.
    Wilson, J. N., and Cooper, P. D. (1963): Aspects of the growth of poliovirus as revealed by the photodynamie effects of neutral red and acridine orange. Virology 21:135–145.PubMedCrossRefGoogle Scholar
  11. 11.
    Kucera, L. S., and Gusdon, J. P. (1976): Transformation of human embryonic fibroblasts by photodynamically inactivated herpes simplex virus, type 2 at supra-optimal temperature. J. Gen. Virol 30:257–261.PubMedCrossRefGoogle Scholar
  12. 12.
    Wallis, C., and Melnick, J. L. (1963): Photodynamie inactivation of poliovirus. Virology 21:332–341.PubMedCrossRefGoogle Scholar
  13. 13.
    Wallis, C., and Melnick, J. L. (1964): Irreversible photosensitization of viruses. Virology 23:520–527.PubMedCrossRefGoogle Scholar
  14. 14.
    Wallis, C., and Melnick, J. L. (1965): Photodynamie inactivation of enteroviruses. J. Bacteriol 89:41–46.PubMedGoogle Scholar
  15. 15.
    Wallis, C., Sakurada, N., and Melnick, J. L. (1963): Influenza vaccine prepared by photodynamie inactivation of virus. J. Immunol 91:677–682.PubMedGoogle Scholar
  16. 16.
    Wallis, C., Scheiris, C., and Melnick, J. L. (1967): Photodynamically inactivated vaccines prepared by growing viruses in cells containing neutral red. J. Immunol 99:1134–1139.PubMedGoogle Scholar
  17. 17.
    Wallis, C., Trulock, S., and Melnick, J. L. (1969): Inherent photosensitivity of herpes virus and other enveloped viruses. J. Gen. Virol. 5:53–61.PubMedCrossRefGoogle Scholar
  18. 18.
    Wallis, C., and Melnick, J. L. (1965): Photodynamic inactivation of animal viruses: a review. Photochem. Photobiol. 4:159–170.CrossRefGoogle Scholar
  19. 19.
    Melnick, J. L., and Wallis, C. (1975): Photodynamic inactivation of herpesvirus. Perspect. Virol. 9:297–314.Google Scholar
  20. 20.
    Felber, T. D., Smith, E. B., Knox, J. M., et al. (1973): Photodynamic inactivation of herpes simplex. J. AMA 223:289–292.Google Scholar
  21. 21.
    Mitchell, J. C., and Stewart, W. B. (1973): Allergic contact dermatitis from neutral red applied for herpes simplex. Arch. Dermatol. 108:689.PubMedCrossRefGoogle Scholar
  22. 22.
    Mitchell, G. A. G., and Buttle, G. A. H. (1943): Proflavine in closed wounds. Lancet 2:749.CrossRefGoogle Scholar
  23. 23.
    Browning, C. H., and Gilmour, W. (1913): Bacterial action and chemical constitution with special reference to basic benzol derivatives. J. Pathol. Bacteriol. 18:144–146.Google Scholar
  24. 24.
    Mcintosh, J., and Selbie, F. R. (1944): Sulphathiazole—proflavine powder in wounds. Lancet 1:591–594.CrossRefGoogle Scholar
  25. 25.
    Wallis, C., Melnick, J. L., and Phillips, C. A. (1965): Bacterialandfungaldecontaminationof virus specimens by differential photosensitization. Am. J. Epidemiol. 81:222–229.PubMedGoogle Scholar
  26. 26.
    Albert, A. (1966): The Acridines, 2nd ed., p. 193. St. Martins Press, New York.Google Scholar
  27. 27.
    Lerman, L. S. (1964): Acridine mutagens and DNA structure. J. Cell. Comp. Physiol. 64(Suppl. 1):1–18.CrossRefGoogle Scholar
  28. 28.
    Gilden, R. V., Kern, J., Lee, Y. K., et al. (1970): Serologic surveys of human cancer patients for antibody to adenovirus T antigens. Am. J. Epidemiol 91:500–509.PubMedGoogle Scholar
  29. 29.
    Simon, M. L. and Vunakis, V. (1962): The photodynamic reaction of methylene blue with deoxyribonucleic acid. J. Mol. Biol. 4:488–499.PubMedCrossRefGoogle Scholar
  30. 30.
    Rapp, F., Li, J. H., and Jerkofsky, M. (1973): Transformation of mammalian cells by DNA-containing viruses following photodynamic inactivation. Virology 55:339–346.PubMedCrossRefGoogle Scholar
  31. 31.
    Li, J. H., Jerkosky, M., and Rapp, F. (1975): Demonstration of oncogenic potential of mammalian cells transformed by DNA-containing viruses following photodynamic inactivation. Int. J. Cancer 15:190–202.PubMedCrossRefGoogle Scholar
  32. 32.
    Melnick, J. L., Courtney, R. J., Powell, K. L., et al. (1976): Studies on herpes simplex virus and cancer. Cancer Res. 36:845–856.PubMedGoogle Scholar
  33. De The, G., Epstein, M. A., and zur Hausen, H. (1975): Oncogenesis and Herpesviruses II: Proceedings of a Symposium Held in Nuremberg, Federal Republic of Germany, 14–16 October 1974. IARC Publications No. 11. Google Scholar
  34. 34.
    Melnick, J. L., Khan, N. C., and Biswal, N. (1977): Photodynamic inactivation of herpes simplex virus and its DNA. Photochem. Photobiol. 25:341–342.PubMedCrossRefGoogle Scholar
  35. 35.
    Khan, N. C., Melnick, J. L., and Biswal, N. (1977): Photodynamic treatment of herpes simplex virus during its replicative cycle. J. Virol. 21:16–23.PubMedGoogle Scholar
  36. 36.
    Dougherty, T. J. (1974): Activated dyes as antitumor agents. J. Natl. Cancer Inst. 52:1333–1336.PubMedGoogle Scholar
  37. 37.
    Tomson, S. H., Emmett, E. A., and Fox, S. H. (1974): Photodestruction of mouse epithelial tumors after oral acridine orange and argon laser. Cancer Res. 34:3124–3127.PubMedGoogle Scholar
  38. 38.
    Seemayer, N. H., and Defendi5V. (1973): Analysis of minimal functions of simian virus 40. II. Enhancement of oncogenic transformation in vitro by UV J. Virol. 12:1265–1271.PubMedGoogle Scholar
  39. 39.
    Seemayer, N. H., Hirai, K., and Defendi, V. (1973): Analysis of minimal functions of simian virus 40. I. Oncogenic transformation of Syrian hamster kidney cells in vitro by photodynamically inactivated SV40. Int. J. Cancer 12:524–531.PubMedCrossRefGoogle Scholar
  40. 40.
    Moore, C., Wallis, C., Melnick, J. L., et al. (1972): Photodynamic treatment of herpes keratitis. Infect. Immun. 5:169–171.PubMedGoogle Scholar
  41. 41.
    Stanley, J. A., and Pinnolis, M. (1976): Light intensity on the photodynamic inactivation of herpes simplex keratitis. Am. J. Ophthalmol. 81:332–336.PubMedGoogle Scholar
  42. 42.
    Friedrich, E. G., Jr. (1973): Relieffor herpes vulvitis. Obstet. Gynecol. 41:74–77.PubMedGoogle Scholar
  43. 43.
    Kaufman, R. H., Gardner, H. L., Brown, D., et al. (1973): Herpes genitalis treated by photodynamic inactivation of virus. Am. J. Obstet. Gynecol. 117:1144–1146.PubMedGoogle Scholar
  44. 44.
    Roome, A. P. C. H., Tinkler, A. E., Hilton, A. L., et al. (1975): Neutral red with photoinactivation in the treatment of herpes genitalis. Br. J. Vener. Dis. 51:130–133.PubMedGoogle Scholar
  45. 45.
    Stanley, J. A. (1973): In: Research to Prevent Blindness. International Science Writers Seminar, pp. 29–30.Google Scholar
  46. 46.
    Editorial (1974): Clinical Dentistry 2:7.Google Scholar
  47. 47.
    Chang, T. -W., and Weinstein, L. (1975): Eczema herpeticum treatment with methylene blue and light. Arch. Dermatol. 111:1174–1175.PubMedCrossRefGoogle Scholar
  48. 48.
    Myers, M. G., Oxman, M. N., Clark, J. E., et al. (1975): Failure of neutral-red photodynamic inactivation in recurrent herpes simplex virus infections. N. Engl. J. Med. 293:945–949.PubMedCrossRefGoogle Scholar
  49. 49.
    Kaufman, R. H., Adam, E., Mirkovic, R. M., et al. (1978): Treatment of genital herpes simplex virus infection with photodynamic inactivation. Am. J. Obstet. Gynecol. 132:861–869.PubMedGoogle Scholar
  50. 50.
    O’Day, D. M., Jones, B. R., Poirier, R., et al. (1975): Proflavine photodynamic viral inactivation in herpes simplex keratitis. Am. J. Ophthalmol. 79:941–948.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • J. L. Melnick
    • 1
  • C. Wallis
    • 1
  1. 1.Department of Virology and EpidemiologyBaylor College of MedicineHoustonUSA

Personalised recommendations