Archives of Pharmacal Research

The Pharmaceutical Society of Korea (대한약학회)
권호 표지

Covalent Linkage of IL-12 and Ovalbumin Confines the Effects of IL-12 to Ovalbumin-specific Immune Responses

  • Published : 1997.10.01
Download PDF
⟨ Previous Next ⟩

Abstract

In order to direct the form of the immune response in an antigen-specific manner, we constructed a fusion protein (OVA/IL12) that contained the T cell-dependent antigen, ovalbumin (OVA), covalently linked to murine interleukin-12 (IL-12). The OVA/IL12 protein was produced in a baculovirus expression system and was purified by anti-OVA immunoaffinity chromatography. The purified OVA/ILI2 protein displayed potent IL-12 bioactivity in an IL-12 proliferation assay. BALB/c mice immunized with the OVA/IL12 protein produced increased quantities of anti-OVA IgG2a antibody compared with mice immunized with recombinant OVA alone. Lymph node cells from the immunized mice with the OVA/IL12 protein produced large amounts of IFN-,Y when restimulated in vitro with OVA, while those from mice immunized with the OVA protein produced little or no IFN-.gamma.. In contrast, immunization with a mixture of OVA and free recombinant IL-12 also induced IFN-.gamma. production, which was not OVA-specific. These studies indicate that the OVA/IL12 fusion protein can induce OVA-specific, Th1-dominated immune responses, and that the covalent linkage of OVA and IL-12 confines the effect of IL-12 to OVA-specific cells.

Keywords

References

  1. Science v.263 The adjuvant effect of interleukin-12 in a vaccine against Leishmania major Afonso,L.C.;Scharton,T.M.;Vieira,L.Q.;Wysocka,M.;Trinchieri,G.;Scott,P.
  2. J. Immunol. v.156 IL-12, as an adjuvant, promotes a T helper 1 cell, but does not suppress a Thelper 2 cell recall response Bliss,J.;Cleace,V.V.;Murray,K.;Wiencis,A.;Ketchum,M.;Maylor,R.;Haire,T.;Resmini,C.;Abbas,A.K.;Wolf,S.
  3. J. Immunol v.153 Idiotype-cytokine fusion proteins as cancer vaccines: relative efficacy of IL-2, IL-4, and granulocyte macrophage colonystimulating fator Chen,T.T.;Tao,M.H.;Levy,R.
  4. Current Protocols in Immunology Coligan,J.E.;Kruisbeek,A.M.;Margulise,D.H.;Shevach,E.M.;Strober,W.
  5. J. Immunol. v.154 IL-12 inhibits IL-4 synthesis in keyhole limpet hemocyanin-primed $CD4^T$ cells through an effect on antigen-presenting cells DeKruyff,R.H.;Fang,Y.;Wolf,S.F.;Umetsu,D.T.
  6. Eur. J. Immunol. v.27 Effect of interleukin-10 on dendritic cell maturation and function De Smedt,T.;Van Mechelen,M.;De Becker,G.;Urbain,J.;Leo,O;Moser,M.
  7. J. Exp. Med. v.183 Interleukin-12 primes human CD4 and CD8 T cell clones for high production of both interferon-γ and interleukin-10 Gerosa,F.;Paganin,C.;Peritt,D.;Paiola,F.;Scupoli,M.T.;Aste-Amezaga,M.;Frank,I.;Trinchieri,G.
  8. Semin. Oncol. v.23 Immunotherapy of cancer with genetically modified tumor vaccines Gilboa,E.
  9. Bioconj. Chem. v.4 Biological activity and in vivo clearance of actitumor antibody/cytokine fusion proteins Gillies,S.D.;Young,D.;Lo,K.M.;Roberts,S.
  10. Eur. J. Immunol. v.26 Interleukin-12 is produced by dendritic cells and mediates T helper 1 development as well as interferon-gamma production by T helper 1 cells Heufler,C.;Koch,F.;Stanzl,U.;Toper,G.;Wysocka,M.;Trinchieri,G.;Enk,A.;Steinman,R.M.;Romani,N.;Schuler,G.
  11. J. Immunol. v.156 IL-12 synergizes with IL-12 and other stimuli in inducing IL-10 production by human T cells Jeannin,P.;Delneste,Y.;Seveso,M.;Life,P.;Bonnefoy,J.Y.
  12. Nucleic Acids Res. v.19 Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus Kaufman,R.J.;Davis,M.V.;Wasley,L.C.;Michnick,D.
  13. J. EXP. Med. v.179 Interleukin-12 induces stable priming for interferon gamma production during differention of human T helper (Th) cells and transient IFN-γ production in established Th2 cell clones Manetti,R.;Gerosa,F.;Giudizi,M.G.;Biagiotti,R.;Parronchi,P.;Piccinni,M.P.;Sampognaro,S.;Maggi,E.;Romagnani,S.;Trinchieri,G.
  14. J. Immunol. v.156 IL-12-induced IL-10 production by human T cells as a negative feedback for IL-12-induced immune responses Meyaard,L.;Hovenkamp,E.;Otto,S.A.;Miedema,F.
  15. Proc. Natl. Acad. Sci. USA v.92 Switch from a type 2 to a type 1 T helper cell response and cure of established Leishmania major infection in mice is induced by combined therapy with interleukin 12 and Pentostam Nabors,G.S.;Afonso,L.C.;Rarrell,J.P.;Scott,P.
  16. J. Immunol. v.126 Monoclonal antibodies to mouse MHC antigens. Ⅲ. Hybridoma antibodies reacting to antigens of the $H-2^b$ haplotype reveal genetic control of isotype expression Ozato,K.;Sachs,D.H.
  17. Blood. v.83 Depletion of B cells in vivo by chimeric mouse human monoclonal antibody to CD20 Reff,M.E.;Carner,K.;Chambers,K.S.;Chinn,P.S.;Leonard,J.E.;Raab,R.;Newman,R.A.;Hanna,N.;Anderson,D.R.
  18. J. Immunol v.151 Anti-TAPA-1 antibodies induce protein tyrosine phosphorylation that is prevented by increasing intracellular thiol levels Schick,M.R.;Ngujen,N.Q.;Levy,S.
  19. J. Leu. Biol. v.59 Immunoregulation by interleukin-12 Trinchieri,G.;Gerosa,F.
  20. J. Immunol v.144 Distinct regulation of lymphokine production is found in fresh versus in vitro primed murine helper T cells. Weinberg,A.D.;English,M.;Swain,S.L.