Skip to main content
NCBI home page
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 Jul;81(14):4515–4519. doi: 10.1073/pnas.81.14.4515

Monoclonal antibodies against murine gamma interferon.

M Prat, G Gribaudo, P M Comoglio, G Cavallo, S Landolfo
PMCID: PMC345621  PMID: 6431408

Abstract

Monoclonal antibodies against murine immune interferon (IFN-gamma) were produced by fusing the murine nonsecreting myeloma cell line P3.X63.Ag8.653 with spleen cells from rats immunized with IFN-gamma-containing supernatants obtained by stimulating a T-cell lymphoma, L12-R4, with phorbol 12-myristate 13-acetate. Supernatants from a twice-cloned hybridoma (AN-18.17.24) were found to neutralize and to adsorb in depletion experiments up to 27 units of mouse IFN-gamma but not equivalent amounts of mouse leukocyte or fibroblast IFNs. The AN-18.17.24 monoclonal antibody neutralized to the same extent mouse IFN-gamma from different sources--namely, (i) concanavalin A-stimulated spleen cells, (ii) alloantigen-stimulated spleen cells, and (iii) monkey fibroblasts transfected with the cloned gene of murine IFN-gamma. Moreover, the monoclonal antibody displayed species specificity, since it did not neutralize IFN-gamma of human origin. Binding inhibition experiments with murine IFN-gamma preparations exposed to enzymatic or physicochemical degradation demonstrated that the protein moiety and not the carbohydrate residues were responsible for the binding to the AN-18.17.24 monoclonal antibody. Finally, this monoclonal antibody immunoprecipitated two molecular species of IFN-gamma of about 16.8 and 17.8 kilodaltons, respectively, from [35S]methionine- or [3H]glucosamine-labeled supernatants of stimulated L12-R4 cells.

Full text

PDF
4515

Images in this article

4518Image
on p.4518

Selected References

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

  1. Gray P. W., Goeddel D. V. Cloning and expression of murine immune interferon cDNA. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5842–5846. doi: 10.1073/pnas.80.19.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gray P. W., Goeddel D. V. Structure of the human immune interferon gene. Nature. 1982 Aug 26;298(5877):859–863. doi: 10.1038/298859a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Gribaudo G., Cofano F., Negro-Ponzi A., Landolfo S. Mouse gamma interferon produced by a cloned T-cell lymphoma. I. Purification and physicochemical characterization. J Interferon Res. 1984;4(1):91–99. doi: 10.1089/jir.1984.4.91. [DOI] [PubMed] [Google Scholar]
  5. Hochkeppel H. K., Menge U., Collins J. Monoclonal antibodies against human fibroblast interferon. Nature. 1981 Jun 11;291(5815):500–501. doi: 10.1038/291500a0. [DOI] [PubMed] [Google Scholar]
  6. Hochkeppel H. K., de Ley M. Monoclonal antibody against human IFN-gamma. Nature. 1982 Mar 18;296(5854):258–259. doi: 10.1038/296258a0. [DOI] [PubMed] [Google Scholar]
  7. Hämmerling G. J., Lemke H., Hämmerling U., Höhmann C., Wallich R., Rajewsky K. Monoclonal antibodies against murine cell surface antigens: anti-H-2, anti-Ia and anti-T cell antibodies. Curr Top Microbiol Immunol. 1978;81:100–106. doi: 10.1007/978-3-642-67448-8_15. [DOI] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Landolfo S., Arnold B., Suzan M. Immune (y) interferon production by murine T cell lymphomas. J Immunol. 1982 Jun;128(6):2807–2809. [PubMed] [Google Scholar]
  10. Landolfo S., Kirchner H., Simon M. M. Production of immune interferon is regulated by more than T cell subset: Lyt-1,2,3 and Qat-5 phenotypes of murine T lymphocytes involved in IFN-gamma production in primary and secondary mixed lymphocyte reaction. Eur J Immunol. 1982 Apr;12(4):295–299. doi: 10.1002/eji.1830120408. [DOI] [PubMed] [Google Scholar]
  11. Landolfo S., Marcucci F., Schirrmacher V., Kirchner H. Characteristics of alloantigens and cellular mechanisms responsible for gamma-interferon production in primary murine MLC. J Interferon Res. 1981;1(3):339–345. doi: 10.1089/jir.1981.1.339. [DOI] [PubMed] [Google Scholar]
  12. Marcucci F., Waller M., Kirchner H., Krammer P. Production of immune interferon by murine T-cell clones from long-term cultures. Nature. 1981 May 7;291(5810):79–81. doi: 10.1038/291079a0. [DOI] [PubMed] [Google Scholar]
  13. McKimm-Breschkin J. L., Mottram P. L., Thomas W. R., Miller J. F. Antigen-specific production of immune interferon by T Cells lines. J Exp Med. 1982 Apr 1;155(4):1204–1209. doi: 10.1084/jem.155.4.1204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Männel D., De Maeyer E., Kemler R., Cachard A., De Maeyer-Guignard J. A rat monoclonal antibody against mouse alpha and beta interferon of all molecular weight species. Nature. 1982 Apr 15;296(5858):664–665. doi: 10.1038/296664a0. [DOI] [PubMed] [Google Scholar]
  15. Otto M. J., Sedmak J. J., Grossberg S. E. Enzymatic modifications of human fibroblast and leukocyte interferons. J Virol. 1980 Aug;35(2):390–399. doi: 10.1128/jvi.35.2.390-399.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prat M., Comoglio P. M. A solid-state competitive binding radioimmunoassay for measurement of antigens solubilized from membranes. J Immunol Methods. 1976;9(3-4):267–272. doi: 10.1016/0022-1759(76)90201-5. [DOI] [PubMed] [Google Scholar]
  17. Prat M., Tarone G., Comoglio P. M. Mouse fibroblasts transformed by Rous sarcoma virus express a virus-specific non-virion transplantation antigen. Int J Cancer. 1981 Jun 15;27(6):797–805. doi: 10.1002/ijc.2910270611. [DOI] [PubMed] [Google Scholar]
  18. Sarzotti M., Landolfo S., Dianzani F. Immune (gamma) interferon production by a murine T cell lymphoma: requirements for macromolecular synthesis and lack of relationship with cell cycle. Infect Immun. 1983 Aug;41(2):624–630. doi: 10.1128/iai.41.2.624-630.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Secher D. S., Burke D. C. A monoclonal antibody for large-scale purification of human leukocyte interferon. Nature. 1980 Jun 12;285(5765):446–450. doi: 10.1038/285446a0. [DOI] [PubMed] [Google Scholar]
  20. Staehelin T., Durrer B., Schmidt J., Takacs B., Stocker J., Miggiano V., Stähli C., Rubinstein M., Levy W. P., Hershberg R. Production of hybridomas secreting monoclonal antibodies to the human leukocyte interferons. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1848–1852. doi: 10.1073/pnas.78.3.1848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vogel S. N., Weedon L. L., Moore R. N., Rosenstreich D. L. Correction of defective macrophage differentiation in C3H/HeJ mice by an interferon-like molecule. J Immunol. 1982 Jan;128(1):380–387. [PubMed] [Google Scholar]
  22. Wietzerbin J., Stefanos S., Lucero M., Falcoff E., O'Malley J. A., Sulkowski E. Physico-chemical characterization and partial purification of mouse immune interferon. J Gen Virol. 1979 Sep;44(3):773–781. doi: 10.1099/0022-1317-44-3-773. [DOI] [PubMed] [Google Scholar]
  23. Wiranowska-Stewart M., Lin L. S., Braude I. A., Stewart W. E., 2nd Production, partial purification and characterization of human and murine interferons--type II. Mol Immunol. 1980 May;17(5):625–633. doi: 10.1016/0161-5890(80)90160-1. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]

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