Abstract
A microassay for interferon is described which uses target cells grown in microculture wells, [3H]uridine to measure vesicular stomatitis virus replication in target cells, and a multiple automated sample harvester to collect the radioactively labeled viral ribonucleic acid onto glass fiber filter disks. The disks were placed in minivials, and radioactivity was counted in a liquid scintillation spectrophotometer. Interferon activity was calculated as the reciprocal of the highest titer which inhibited the incorporation of [3H]uridine into viral ribonucleic acid by 50%. Interferon titers determined by the microassay were similar to the plaque reduction assay when 100 plaque-forming units of challenge vesicular stomatitis virus was used. However, it was found that the interferon titers decreased approximately 2-fold for each 10-fold increase in the concentration of challenge vesicular stomatitis virus when tested in the range of 10(2) to 10(5) plaque-forming units. Interferon titers determined by the microassay show a high degree of repeatability, and the assay can be used to measure small and large numbers of interferon samples.
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- Allen P. T., Giron D. J. Rapid sensitive assay for interferons based on the inhibition of MM virus nucleic acid synthesis. Appl Microbiol. 1970 Sep;20(3):317–322. doi: 10.1128/am.20.3.317-322.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campbell J. B., Grunberger T., Kochman M. A., White S. L. A microplaque reduction assay for human and mouse interferon. Can J Microbiol. 1975 Aug;21(8):1247–1253. doi: 10.1139/m75-186. [DOI] [PubMed] [Google Scholar]
- FINTER N. B. QUANTITATIVE HAEMADSORPTION, A NEW ASSAY TECHNIQUE. I. ASSAY OF INTERFERON. Virology. 1964 Dec;24:589–597. doi: 10.1016/0042-6822(64)90212-0. [DOI] [PubMed] [Google Scholar]
- Gauntt C. J. Effect of interferon on synthesis of ssRNA in reovirus type 3-infected L cell cultures. Biochem Biophys Res Commun. 1972 Jun 9;47(5):1228–1236. doi: 10.1016/0006-291x(72)90966-7. [DOI] [PubMed] [Google Scholar]
- Green J. A. Rapid assay of interferon. Tex Rep Biol Med. 1977;35:167–172. [PubMed] [Google Scholar]
- Knudsen R. C., Ahmed A. A., Sell K. W. An in vitro microassay for lymphotoxin using microculture plates and the multiple automated sample harvester. J Immunol Methods. 1974 May;5(1):55–63. doi: 10.1016/0022-1759(74)90045-3. [DOI] [PubMed] [Google Scholar]
- Knudsen R. C., Callahan L. T., 3rd, Ahmed A., Sell K. W. Use of microculture plates and the multiple automated sample harvester for in vitro microassay of bacterial toxins. Appl Microbiol. 1974 Aug;28(2):326–327. doi: 10.1128/am.28.2.326-327.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koblet H., Kohler U., Wyler R. Optimization of the interferon assay using inhibition of Semliki forest virus-ribonucleic acid synthesis. Appl Microbiol. 1972 Sep;24(3):323–327. doi: 10.1128/am.24.3.323-327.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McLaren C. Influence of cell age on production and assay of mouse interferon in L-cells. Arch Gesamte Virusforsch. 1970;32(1):13–18. doi: 10.1007/BF01241514. [DOI] [PubMed] [Google Scholar]
- McWilliams M., Finkelstein M. S., Allen P. T., Giron D. J. Assay of chick interferons by the inhibition of viral ribonucleic acid synthesis. Appl Microbiol. 1971 May;21(5):959–961. doi: 10.1128/am.21.5.959-961.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mirchamsy H., Rapp F. A new overlay for plaquing animal viruses. Proc Soc Exp Biol Med. 1968 Oct;129(1):13–17. doi: 10.3181/00379727-129-33237. [DOI] [PubMed] [Google Scholar]
- Richmond J. Y., Polatnick J. Further studies of the physical and metabolic properties of foot-and-mouth disease virus temperature-sensitive mutants. Infect Immun. 1976 May;13(5):1392–1396. doi: 10.1128/iai.13.5.1392-1396.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Richmond J. Y. Preadsorption of boar semen with kaolin: increased efficiency of foot-and-mouth disease virus detection. Am J Vet Res. 1978 Oct;39(10):1612–1616. [PubMed] [Google Scholar]
- SHATKIN A. J. Actinomycin inhibition of ribonucleic acid synthesis and poliovirus infection of HeLa cells. Biochim Biophys Acta. 1962 Aug 20;61:310–313. doi: 10.1016/0926-6550(62)90095-6. [DOI] [PubMed] [Google Scholar]
- Strong D. M., Ahmed A. A., Thurman G. B., Sell K. W. In vitro stimulation of murine spleen cells using a microculture system and a multiple automated sample harvester. J Immunol Methods. 1973 Apr;2(3):279–291. doi: 10.1016/0022-1759(73)90054-9. [DOI] [PubMed] [Google Scholar]
- Suzuki J., Akaboshi T., Kobayashi S. A rapid and simple method for assaying interferon. Jpn J Microbiol. 1974 Nov;18(6):449–456. doi: 10.1111/j.1348-0421.1974.tb00833.x. [DOI] [PubMed] [Google Scholar]
- Thurman G. B., Strong D. M., Ahmed A., Green S. S., Sell K. W., Hartzman R. J., Bach F. H. Human mixed lymphocyte cultures. Evaluation of a microculture technique utilizing the Multiple Automated Sample Harvester (MASH). Clin Exp Immunol. 1973 Oct;15(2):289–302. [PMC free article] [PubMed] [Google Scholar]
- Tilles J. G., Finland M. Microassay for human and chick cell interferons. Appl Microbiol. 1968 Nov;16(11):1706–1707. doi: 10.1128/am.16.11.1706-1707.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vassef A., Beaud G., Paucker K., Lengyel P. Interferon assay based on the inhibition of double-stranded reovirus RNA accumulation in mouse L cells. J Gen Virol. 1973 Apr;19(1):81–87. doi: 10.1099/0022-1317-19-1-81. [DOI] [PubMed] [Google Scholar]
- Woody J. N., Ahmed A., Knudsen R. C., Strong D. M., Sell K. W. Human T-cell heterogeneity as delineated with a specific human thymus lymphocyte antiserum. In vitro effects on mitogen response mixed leukocyte culture, cell-mediated lymphocytotoxicity, and lymphokine production. J Clin Invest. 1975 May;55(5):956–966. doi: 10.1172/JCI108025. [DOI] [PMC free article] [PubMed] [Google Scholar]