Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1985 May;27(5):802–805. doi: 10.1128/aac.27.5.802

Determination of inhibitory concentrations of antiviral agents in cell culture by use of an enzyme immunoassay with virus-specific, peroxidase-labeled monoclonal antibodies.

F H van Tiel, W A Boere, T Harmsen, C A Kraaijeveld, H Snippe
PMCID: PMC180155  PMID: 3925876

Abstract

An enzyme immunoassay (EIA) to determine 50% inhibitory concentrations of drugs which suppress Semliki Forest virus replication is described. Inhibition of virus replication was measured in L-cells, seeded as monolayers in 96-well plates by use of horseradish peroxidase-labeled monoclonal antibodies directed against the E1 glycoprotein of Semliki Forest virus. The antiviral agents tested were cycloheximide, tunicamycin, NH4Cl, and disodium cromoglycate. The 50% inhibitory concentration of these antiviral agents was arbitrarily defined as the concentration of drug, in culture medium, associated with 50% reduction of the control absorbance value measured on Semliki Forest virus-infected cells without drug in the culture fluid. Twenty-two hours after infection the 50% inhibitory concentrations of the drugs were 0.2 microgram/ml for cycloheximide, 0.8 microgram/ml for tunicamycin, 0.3 mg/ml for NH4Cl, and 4.9 mg/ml for disodium cromoglycate. These values are similar to those determined by others with conventional methods of virus quantification. This test is sensitive and easy to perform and therefore is suited for large-scale experiments.

Full text

PDF
802

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baglioni C., Vesco C., Jacobs-Lorena M. The role of ribosomal subunits in mammalian cells. Cold Spring Harb Symp Quant Biol. 1969;34:555–565. doi: 10.1101/sqb.1969.034.01.063. [DOI] [PubMed] [Google Scholar]
  2. Bauer D. J., Selway J. W., Batchelor J. F., Tisdale M., Caldwell I. C., Young D. A. 4',6-Dichloroflavan (BW683C), a new anti-rhinovirus compound. Nature. 1981 Jul 23;292(5821):369–370. doi: 10.1038/292369a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Becker Y., Hadar J. Antivirals 1980--an update. Prog Med Virol. 1980;26:1–44. [PubMed] [Google Scholar]
  4. Boere W. A., Benaissa-Trouw B. J., Harmsen M., Kraaijeveld C. A., Snippe H. Neutralizing and non-neutralizing monoclonal antibodies to the E2 glycoprotein of Semliki Forest virus can protect mice from lethal encephalitis. J Gen Virol. 1983 Jun;64(Pt 6):1405–1408. doi: 10.1099/0022-1317-64-6-1405. [DOI] [PubMed] [Google Scholar]
  5. Boere W. A., Harmsen T., Vinjé J., Benaissa-Trouw B. J., Kraaijeveld C. A., Snippe H. Identification of distinct antigenic determinants on Semliki Forest virus by using monoclonal antibodies with different antiviral activities. J Virol. 1984 Nov;52(2):575–582. doi: 10.1128/jvi.52.2.575-582.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Foreman J. C., Garland L. G. Cromoglycate and other antiallergic drugs: a possible mechanism of action. Br Med J. 1976 Apr 3;1(6013):820–821. doi: 10.1136/bmj.1.6013.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gadler H. Nucleic acid hybridization for measurement of effects of antiviral compounds on human cytomegalovirus DNA replication. Antimicrob Agents Chemother. 1983 Sep;24(3):370–374. doi: 10.1128/aac.24.3.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HAFF R. F. INHIBITION OF THE MULTIPLICATION OF PSEUDORABIES VIRUS BY CYCLOHEXAMIDE. Virology. 1964 Mar;22:430–431. doi: 10.1016/0042-6822(64)90036-4. [DOI] [PubMed] [Google Scholar]
  9. Helenius A., Marsh M., White J. Inhibition of Semliki forest virus penetration by lysosomotropic weak bases. J Gen Virol. 1982 Jan;58(Pt 1):47–61. doi: 10.1099/0022-1317-58-1-47. [DOI] [PubMed] [Google Scholar]
  10. Henderson B. E., Metselaar D., Kirya G. B., Timms G. L. Investigations into yellow fever virus and other arboviruses in the northern regions of Kenya. Bull World Health Organ. 1970;42(5):787–795. [PMC free article] [PubMed] [Google Scholar]
  11. Klenk H. D., Schwarz R. T. Viral glycoprotein metabolism as a target for antiviral substances. Antiviral Res. 1982 Sep;2(4):177–190. doi: 10.1016/0166-3542(82)90041-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kraaijeveld C. A., Harmsen M., Khader Boutahar-Trouw B. Delayed-type hypersensitivity against Semliki Forest virus in mice. Infect Immun. 1979 Feb;23(2):219–223. doi: 10.1128/iai.23.2.219-223.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Loveday D. E., Wenham R. B. Effect of disodium cromoglycate on virus-cell systems in vitro. Br Med J. 1977 Aug 27;2(6086):557–558. doi: 10.1136/bmj.2.6086.557-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
  15. Penttinen K., Aarnio A., Hovi T. Disodium cromoglycate can inhibit virus-induced cytopathic effects in vitro. Br Med J. 1977 Jan 8;1(6053):82–82. doi: 10.1136/bmj.1.6053.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schwarz R. T., Rohrschneider J. M., Schmidt M. F. Suppression of glycoprotein formation of Semliki Forest, influenza, and avian sarcoma virus by tunicamycin. J Virol. 1976 Sep;19(3):782–791. doi: 10.1128/jvi.19.3.782-791.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wahren B., Harmenberg J., Sundqvist V. A., Levén B., Sköldenberg B. A novel method for determining the sensitivity of herpes simplex virus to antiviral compounds. J Virol Methods. 1983 Mar;6(3):141–149. doi: 10.1016/0166-0934(83)90026-5. [DOI] [PubMed] [Google Scholar]
  18. van Tiel F. H., Boere W. A., Vinjé J., Harmsen T., Benaissa-Trouw B. J., Kraaijeveld C. A., Snippe H. Detection of Semliki Forest virus in cell culture by use of an enzyme immunoassay with peroxidase-labeled monoclonal antibodies specific for glycoproteins E1 and E2. J Clin Microbiol. 1984 Sep;20(3):387–390. doi: 10.1128/jcm.20.3.387-390.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES