Monoclonal antibodies against murine γ interferon

M. Prat, G. Gribaudo, P. M. Comoglio, G. Cavallo, S. Landolfo

Research output: Contribution to journalArticlepeer-review

Abstract

Monoclonal antibodies against murine immune interferon (IFN-γ) were produced by fusing the murine nonsecreting myeloma cell line P3.X63.Ag8.653 with spleen cells from rats immunized with IFN-γ-containing supernatants obtained by stimulating a T-cell lymphoma, L12-R4, with phorbol 12-myristate 13-acetate. Supernatants from a twice-cloned hydridoma (AN-18.17.24) were found to neutralize and to absorb in depletion experiments up to 27 units of mouse IFN-γ but not equivalent amounts of mouse leukocyte or fibroblast IFNs. The AN-18.17.24 monoclonal antibody neutralized to the same extent mouse IFN-γ 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-γ. Moreover, the monoclonal antibody displayed species specificity, since it did not neutralize IFN-γ of human origin. Binding inhibition experiments with murine IFN-γ 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-γ of about 16.8 and 17.8 kilodaltons, respectively, from [35S]methionine- or [3H]glucosamine-labeled supernatants of stimulated L12-R4 cells.

Original languageEnglish
Pages (from-to)4515-4519
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number14 I
Publication statusPublished - 1984

ASJC Scopus subject areas

  • Genetics
  • General

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