Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Aug;81(16):5219–5222. doi: 10.1073/pnas.81.16.5219

Use of monoclonal antibodies as sensitive and specific probes for biologically active human gamma-interferon.

T W Chang, S McKinney, V Liu, P C Kung, J Vilcek, J Le
PMCID: PMC391669  PMID: 6433346

Abstract

Mouse monoclonal antibodies B1 and B3 are specific for natural and Escherichia coli-derived recombinant human gamma-interferon (IFN-gamma). The two antibodies recognize different epitopes of the IFN-gamma molecule and do not compete with each other's binding. We have used these two antibodies to construct a solid-phase, sandwich immunoradiometric assay for human IFN-gamma. Purified antibody B1 was coated on polystyrene beads (0.64 cm in diameter) and used as the solid-phase immunoadsorbent and antibody B3 was labeled with 125I and used as tracer. This assay can be completed in about 4 hr and is capable of detecting IFN-gamma levels in human serum or tissue culture fluids as low as 0.1 NIH reference unit/ml. Recombinant human IFN-gamma derived from E. coli was detectable at a concentration of 0.02 ng/ml. The assay appears to be specific for the biologically active forms of IFN-gamma, since after exposure to pH 2, 37 degrees C, or 56 degrees C, biological activity and reactivity in the immunoradiometric assay decreased in parallel. The immunoradiometric assay can be employed for the analysis of the structural characteristics of the human IFN-gamma molecule.

Full text

PDF
5219

Selected References

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

  1. Basham T. Y., Merigan T. C. Recombinant interferon-gamma increases HLA-DR synthesis and expression. J Immunol. 1983 Apr;130(4):1492–1494. [PubMed] [Google Scholar]
  2. Chang T. W., Testa D., Kung P. C., Perry L., Dreskin H. J., Goldstein G. Cellular origin and interactions involved in gamma-interferon production induced by OKt3 monoclonal antibody. J Immunol. 1982 Feb;128(2):585–589. [PubMed] [Google Scholar]
  3. Cunningham A. L., Merigan T. C. gamma Interferon production appears to predict time of recurrence of herpes labialis. J Immunol. 1983 May;130(5):2397–2400. [PubMed] [Google Scholar]
  4. Ennis F. A., Meager A. Immune interferon produced to high levels by antigenic stimulation of human lymphocytes with influenza virus. J Exp Med. 1981 Nov 1;154(5):1279–1289. doi: 10.1084/jem.154.5.1279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fujii N., Minagawa T., Nakane A., Kato F., Ohno S. Spontaneous production of gamma-interferon in cultures of T lymphocytes obtained from patients with Behçet's disease. J Immunol. 1983 Apr;130(4):1683–1686. [PubMed] [Google Scholar]
  6. Gray P. W., Leung D. W., Pennica D., Yelverton E., Najarian R., Simonsen C. C., Derynck R., Sherwood P. J., Wallace D. M., Berger S. L. Expression of human immune interferon cDNA in E. coli and monkey cells. Nature. 1982 Feb 11;295(5849):503–508. doi: 10.1038/295503a0. [DOI] [PubMed] [Google Scholar]
  7. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  8. Havell E. A., Vilcek J. Production of high-titered interferon in cultures of human diploid cells. Antimicrob Agents Chemother. 1972 Dec;2(6):476–484. doi: 10.1128/aac.2.6.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Herberman R. B., Ortaldo J. R., Mantovani A., Hobbs D. S., Kung H. F., Pestka S. Effect of human recombinant interferon on cytotoxic activity of natural killer (NK) cells and monocytes. Cell Immunol. 1982 Feb;67(1):160–167. doi: 10.1016/0008-8749(82)90208-8. [DOI] [PubMed] [Google Scholar]
  10. Hooks J. J., Jordan G. W., Cupps T., Moutsopoulos H. M., Fauci A. S., Notkins A. L. Multiple interferons in the circulation of patients with systemic lupus erythematosus and vasculitis. Arthritis Rheum. 1982 Apr;25(4):396–400. doi: 10.1002/art.1780250406. [DOI] [PubMed] [Google Scholar]
  11. Johnson H. M., Stanton G. J., Baron S. Relative ability of mitogens to stimulate production of interferon by lymphoid cells and to induce suppression of the in vitro immune response. Proc Soc Exp Biol Med. 1977 Jan;154(1):138–141. [PubMed] [Google Scholar]
  12. Le J., Barrowclough B. S., Vilcek J. Monoclonal antibodies to human immune interferon and their application for affinity chromatography. J Immunol Methods. 1984 Apr 13;69(1):61–70. doi: 10.1016/0022-1759(84)90277-1. [DOI] [PubMed] [Google Scholar]
  13. Le J., Prensky W., Yip Y. K., Chang Z., Hoffman T., Stevenson H. C., Balazs I., Sadlik J. R., Vilcek J. Activation of human monocyte cytotoxicity by natural and recombinant immune interferon. J Immunol. 1983 Dec;131(6):2821–2826. [PubMed] [Google Scholar]
  14. Le J., Rubin B. Y., Kelker H. C., Feit C., Nagler C., Vilcek J. Natural and recombinant Escherichia coli-derived interferon-gamma differ in their reactivity with monoclonal antibody. J Immunol. 1984 Mar;132(3):1300–1304. [PubMed] [Google Scholar]
  15. Le J., Vilcek J. Lymphokine-mediated activation of human monocytes: neutralization by monoclonal antibody to interferon-gamma. Cell Immunol. 1984 Apr 15;85(1):278–283. doi: 10.1016/0008-8749(84)90299-5. [DOI] [PubMed] [Google Scholar]
  16. Nakamura M., Manser T., Pearson G. D., Daley M. J., Gefter M. L. Effect of IFN-gamma on the immune response in vivo and on gene expression in vitro. 1984 Jan 26-Feb 1Nature. 307(5949):381–382. doi: 10.1038/307381a0. [DOI] [PubMed] [Google Scholar]
  17. Nathan C. F., Murray H. W., Wiebe M. E., Rubin B. Y. Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J Exp Med. 1983 Sep 1;158(3):670–689. doi: 10.1084/jem.158.3.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Novick D., Eshhar Z., Fischer D. G., Friedlander J., Rubinstein M. Monoclonal antibodies to human interferon-gamma: production, affinity purification and radioimmunoassay. EMBO J. 1983;2(9):1527–1530. doi: 10.1002/j.1460-2075.1983.tb01618.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ohno S., Kato F., Matsuda H., Fujii N., Minagawa T. Detection of gamma interferon in the sera of patients with Behçet's disease. Infect Immun. 1982 Apr;36(1):202–208. doi: 10.1128/iai.36.1.202-208.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pestka S., Kelder B., Langer J. A., Staehelin T. Monoclonal antibodies can discriminate between some active and inactive forms of leukocyte interferon. Arch Biochem Biophys. 1983 Jul 1;224(1):111–116. doi: 10.1016/0003-9861(83)90194-7. [DOI] [PubMed] [Google Scholar]
  21. Rhodes-Feuillette A., Druilhe P., Canivet M., Gentilini M., Périés J. Présence d'interféron circulant dans le sérum de malades infectés par Plasmodium falciparum. C R Seances Acad Sci III. 1981 Nov 23;293(11):635–637. [PubMed] [Google Scholar]
  22. Rinderknecht E., O'Connor B. H., Rodriguez H. Natural human interferon-gamma. Complete amino acid sequence and determination of sites of glycosylation. J Biol Chem. 1984 Jun 10;259(11):6790–6797. [PubMed] [Google Scholar]
  23. Roberts W. K., Vasil A. Evidence for the identity of murine gamma interferon and macrophage activating factor. J Interferon Res. 1982;2(4):519–532. doi: 10.1089/jir.1982.2.519. [DOI] [PubMed] [Google Scholar]
  24. Rubin B. Y., Bartal A. H., Anderson S. L., Millet S. K., Hirshaut Y., Feit C. The anticellular and protein-inducing activities of human gamma interferon preparations are mediated by the interferon. J Immunol. 1983 Mar;130(3):1019–1020. [PubMed] [Google Scholar]
  25. Schreiber R. D., Pace J. L., Russell S. W., Altman A., Katz D. H. Macrophage-activating factor produced by a T cell hybridoma: physiochemical and biosynthetic resemblance to gamma-interferon. J Immunol. 1983 Aug;131(2):826–832. [PubMed] [Google Scholar]
  26. Steeg P. S., Johnson H. M., Oppenheim J. J. Regulation of murine macrophage Ia antigen expression by an immune interferon-like lymphokine: inhibitory effect of endotoxin. J Immunol. 1982 Dec;129(6):2402–2406. [PubMed] [Google Scholar]
  27. Targan S., Stebbing N. In vitro interactions of purified cloned human interferons on NK cells: enhanced activation. J Immunol. 1982 Sep;129(3):934–935. [PubMed] [Google Scholar]
  28. Trinchieri G., Santoli D. Anti-viral activity induced by culturing lymphocytes with tumor-derived or virus-transformed cells. Enhancement of human natural killer cell activity by interferon and antagonistic inhibition of susceptibility of target cells to lysis. J Exp Med. 1978 May 1;147(5):1314–1333. doi: 10.1084/jem.147.5.1314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wheelock E. F. Interferon-like virus-inhibitor induced in human leukocytes by phytohemagglutinin. Science. 1965 Jul 16;149(3681):310–311. [PubMed] [Google Scholar]
  30. Yip Y. K., Barrowclough B. S., Urban C., Vilcek J. Molecular weight of human gamma interferon is similar to that of other human interferons. Science. 1982 Jan 22;215(4531):411–413. doi: 10.1126/science.6173921. [DOI] [PubMed] [Google Scholar]
  31. Yip Y. K., Barrowclough B. S., Urban C., Vilcek J. Purification of two subspecies of human gamma (immune) interferon. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1820–1824. doi: 10.1073/pnas.79.6.1820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ziola B. R., Matikainen M. T., Salmi A. Polystyrene balls as the solid-phase of a double-antibody radioimmunoassay for human serum albumin. J Immunol Methods. 1977;17(3-4):309–317. doi: 10.1016/0022-1759(77)90113-2. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES