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  • Nonviral Transfer Technology
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Regulation of T-helper-1 versus T-helper-2 activity and enhancement of tumor immunity by combined DNA-based vaccination and nonviral cytokine gene transfer

Gene Therapy volume 7, pages 481–492 (2000)Cite this article

Abstract

Intramuscular (i.m.) injections of a plasmid encoding human carcinoembryonic antigen (CEA) elicited both humoral and cellular immune responses in mice, but only partial inhibition of the growth of transplanted syngeneic CEA-positive P815 tumor cells (CEA/P815). Coinjection of the CEA vector with a vector encoding either interferon-γ (IFNγ) or IL-12 promoted IgG2a isotype anti-CEA antibody production, anti-CEA/P815 CTL activity and greater resistance to CEA/P815 tumor challenge. As well, CEA/P815-stimulated IFNγ secretion in vitro was increased, but IL-4 diminished, consistent with a T-helper type 1 (Th1) response. In contrast, coinjection of the CEA vector with an IL-4 vector increased IgG1 production, but reduced CTL activity and resistance to tumor challenge. The latter treatment inhibited CEA/P815-dependent IFNγ production but enhanced IL-4 secretion, consistent with a Th type 2 (Th2) response. Antitumor immunity was enhanced when the CEA and IL-12 plasmids were coinjected at the same muscle site, but not at separate sites despite increased serum IL-12 levels. Though the tumor cells expressed neomycin phosphotransferase, mice immunized with vectors encoding that protein (without CEA) were not protected against tumor growth, and produced no CTLs except for low levels when coinjected with an IL-12 vector. Thus, we show that immunity elicited by DNA vaccination against CEA can be biased to a protective type (high Th1 and CTL activity) or nonprotective type (high Th2 and low CTL activity) by i.m. coinjection of cytokine-expressing plasmids. IL-12 appears to act locally, but not systemically, through an adjuvant effect.

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References

  1. Cohen AD, Boyer JD, Paul DB . Modulating the immune response to genetic immunization FASEB J 1998 12: 1611–1626

    Article  CAS  PubMed  Google Scholar 

  2. Rock KL . A new foreign policy: MHC class I molecules monitor the outside world Immunol Today 1996 17: 131–138

    Article  CAS  PubMed  Google Scholar 

  3. Jondal M, Schirmbeck R, Reimann J . MHC class I-restricted CTL responses to exogenous antigens Immunity 1996 5: 295–302

    Article  CAS  PubMed  Google Scholar 

  4. Raychaudhuri S, Morrow WJ . Can soluble antigen induce CD8+ cytotoxic T cell responses? A paradox revisited Immunol Today 1993 14: 344–348

    Article  CAS  PubMed  Google Scholar 

  5. Engers HD et al. Functional activity in vivo of effector T cell populations J Immunol 1984 133: 1664–1670

    CAS  PubMed  Google Scholar 

  6. Yee C, Riddell SR, Greenberg PD . Prospects for adoptive T cell therapy Curr Opin Immunol 1997 9: 705–708

    Article  Google Scholar 

  7. Carmichael A, Jin X, Sissons P, Borysiewicz L . Quantitative analysis of human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocyte (CTL) response at different stages of HIV-1 infection: differential CTL responses to HIV-1 and Epstein–Barr virus in late disease J Exp Med 1993 177: 249–256

    Article  CAS  PubMed  Google Scholar 

  8. Böhm W et al. T cell-mediated, IFNγ-facilitated rejection of murine B16 melanomas J Immunol 1998 161: 897–908

    PubMed  Google Scholar 

  9. Xu D, Liew FY . Protection against leishmaniasis by injection of DNA encoding a major surface glycoprotein, gp63, of 1-major Immunol 1995 84: 173–176

    CAS  Google Scholar 

  10. Boyle JS, Koniaras C, Lew AM . Influence of cellular location and expressed antigen on the efficacy of DNA vaccination: cytotoxic T lymphocyte and antibody responses are suboptimal when antigen is cytoplasmic after intramuscular DNA injection Int Immunol 1997 9: 1897–1906

    Article  CAS  PubMed  Google Scholar 

  11. Schirmbeck R, Böhm W, Reimann J . DNA vaccination primes MHC class I-restricted, Simian virus 40 large tumor antigen-specific CTL in H-2d mice that reject syngeneic tumors J Immunol 1996 157: 3550–3558

    CAS  PubMed  Google Scholar 

  12. Thompson J, Zimmermann W . The carcinoembryonic antigen gene family: structure, expression and evolution Tumor Biol 1988 9: 63–83

    Article  CAS  Google Scholar 

  13. Thompson JA, Grunert F, Zimmermann W . Carcinoembryonic antigen gene family: molecular biology and clinical perspectives J Clin Lab Anal 1991 5: 344–366

    Article  CAS  PubMed  Google Scholar 

  14. Hodge JW . Carcinoembryonic antigen as a target for cancer vaccines Cancer Immunol Immunother 1996 43: 127–134

    Article  CAS  PubMed  Google Scholar 

  15. Hunter RL, Olsen MR, Bennett B . Copolymer adjuvants and titermax. In: Stewart-Tull DES (ed) The Theory and Practical Application of Adjuvants John Wiley: Chichester 1995 pp 51–94

    Google Scholar 

  16. Allsopp CE et al. Comparison of numerous delivery systems for the induction of cytotoxic T lymphocytes by immunization Eur J Immunol 1996 25: 1137–1140

    Google Scholar 

  17. Ishii KJ et al. Activity and safety of DNA plasmids encoding IL-4 and IFNgamma Gene Therapy 1999 6: 237–244

    Article  CAS  PubMed  Google Scholar 

  18. Sin JI et al. In vivo modulation of vaccine-induced immune responses toward a Th1 phenotype increases potency and vaccine effectiveness in a herpes simplex virus type 2 mouse model J Virol 1999 73: 501–509

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Kim JJ et al. Modulation of amplitude and direction of in vivo immune responses by co-administration of cytokine gene expression cassettes with DNA immunogens Euro J Immunol 1998 28: 1089–1103

    Article  CAS  Google Scholar 

  20. Chow YH et al. Development of Th1 and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccines can be modulated by codelivery of various cytokine genes J Immunol 1998 160: 1320–1329

    CAS  PubMed  Google Scholar 

  21. Conry RM et al. A carcinoembryonic antigen polynucleotide vaccine has in vivo antitumor activity Gene Therapy 1995 2: 59–65

    CAS  PubMed  Google Scholar 

  22. Coffman RL, Lebman DA, Rothman P . Mechanism and regulation of immunoglobulin isotype switching Adv Immunol 1993 54: 229–270

    Article  CAS  PubMed  Google Scholar 

  23. Bloom ET, Horvath JA . Cellular and molecular mechanisms of the IL-12-induced increase in allospecific murine cytolytic T cell activity: implications for the age-related decline in CTL J Immunol 1994 152: 4242–4254

    CAS  PubMed  Google Scholar 

  24. Seder RA, Gassinelli R, Sher A, Paul WE . IL-12 acts directly on CD4+ T cells to enhance priming for IFNγ production and diminishes IL-4 inhibition in such priming Proc Natl Acad Sci USA 1993 90: 10188–10192

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Abbas AK, Murphy KM, Sher A . Functional diversity of helper T lymphocytes Nature 1996 383: 787–793

    Article  CAS  PubMed  Google Scholar 

  26. Tahara H, Lotze MT . Antitumor effects of interleukin-12 (IL-12): applications for the immunotherapy and gene therapy of cancer Gene Therapy 1995 2: 96–106

    CAS  PubMed  Google Scholar 

  27. Schultz J et al. Long-lasting anti-metastatic efficiency of interleukin 12-encoding plasmid DNA Hum Gene Ther 1999 10: 407–417

    Article  CAS  PubMed  Google Scholar 

  28. Fagerberg J et al. Recombinant carcinoembryonic antigen and granulocyte–macrophage colony-stimulating factor for active immunization of colorectal carcinoma patients. Proceedings of the 11th Annual Meeting of the Society for Biological Therapy, Madison, WI: Society for Biological Therapy, 1996

    Article  Google Scholar 

  29. Irvine K, Kantor J, Schlom J . Comparison of a CEA-recombinant vaccinia virus, purified CEA and an anti-idiotype bearing the image of a CEA epitope in the treatment and prevention of CEA-expressing tumors Vaccine Res 1993 2: 79–94

    CAS  Google Scholar 

  30. Bei R et al. Enhanced immune responses and anti-tumor activity by baculovirus recombinant carcinoembryonic antigen (CEA) in mice primed with the recombinant vaccinia CEA J Immunother 1994 16: 275–282

    Article  CAS  Google Scholar 

  31. Kantor J et al. Antitumor activity and immune responses induced by a recombinant carcinoembryonic antigen-vaccinia virus vaccine J Natl Can Inst 1992 84: 1084–1091

    Article  CAS  Google Scholar 

  32. McLaughlin JP, Schlom J, Kontor JA, Greiner JW . Improved immunotherapy of a recombinant carcinoembryonic antigen vaccinia vaccine when given in combination with interleukin-2 Cancer Res 1996 56: 2361–2367

    CAS  PubMed  Google Scholar 

  33. Bei R et al. The use of a cationic liposome formulation (DOTAP) mixed with a recombinant tumor-associated antigen to induce immune responses and protective immunity in mice J Immunother 1998 21: 159–169

    Article  CAS  PubMed  Google Scholar 

  34. Tsuji T et al. Enhancement of cell-mediated immunity against HIV-1 induced by coinoculation of plasmid-encoded HIV-1 antigen with plasmid expressing IL-12 J Immunol 1997 158: 4008–4013

    CAS  PubMed  Google Scholar 

  35. Kim JJ et al. In vivo engineering of a cellar immune response by coadministration of IL-12 expression vector with a DNA immunogen J Immunol 1997 158: 816–826

    CAS  PubMed  Google Scholar 

  36. Michael G, Gesien A, Tokushige K, Wands JR . Enhancement of cellular and humoral immune responses to hepatitis C virus core protein using DNA-based vaccines augmented with cytokine-expressing plasmids J Immunol 1997 158: 1231–1237

    Google Scholar 

  37. OGarra A . Cytokines induce the development of functionally heterogeneous T helper cell subsets Immunity 1998 8: 275–283

    Article  CAS  Google Scholar 

  38. Sgadari C, Angiolillo AL, Tosato G . Inhibition of angiogenesis by interleukin-12 is mediated by the interferon-inducible protein 10 Blood 1996 87–9: 3877–3882

    Google Scholar 

  39. Macatonia SE, Hsieh C, Murphy KM, O'Garra A . Dendritic cells and macrophages are required for Th1development of CD4+ T cells from αβ-TCR transgenic mice: IL-12 substitution for macrophages to stimulate IFNγ production is IFNγ-dependent Int Immunol 1993 5: 1119–1128

    Article  CAS  PubMed  Google Scholar 

  40. Kobayashi M et al. Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes J Exp Med 1989 170: 827–845

    Article  CAS  PubMed  Google Scholar 

  41. Chatelain R, Varkila K, Coffman RL . IL-4 induces a Th2 response in Leishmania major-infected mice J Immunol 1992 148: 1182–1187

    CAS  PubMed  Google Scholar 

  42. Seder RA et al. The presence of interleukin 4 during in vitro priming determines the lymphokine-producing potential of CD4+ T cells from T cell receptor transgenic mice J Exp Med 1992 176: 1091–1098

    Article  CAS  PubMed  Google Scholar 

  43. Liew FY et al. Macrophage activation by interferon-gamma from host-protective T cells is inhibited by interleukin (IL)3 and IL4 produced by disease-promoting T cells in leishmaniasis Eur J Immunol 1989 19: 1227–1232

    Article  CAS  PubMed  Google Scholar 

  44. Paul WE, Ohara J . B-cell stimulatory factor-1/interleukin-4 Annu Rev Immunol 1987 5: 429–459

    Article  CAS  PubMed  Google Scholar 

  45. Nastala CL et al. Recombinant IL-12 administration induces tumor regression in association with IFNγ production J Immunol 1994 153: 1697–1706

    CAS  PubMed  Google Scholar 

  46. Zitvogel L et al. Cancer immunotherapy of established tumors with IL-12. Effective delivery by genetically engineered fibroblasts J Immunol 1995 155: 1393–1403

    CAS  PubMed  Google Scholar 

  47. Martinotti A et al. CD4 T cells inhibit in vivo the CD8-mediated immune response against murine colon carcinoma cells transduced with interleukin-12 genes Eur J Immunol 1995 25: 137–146.

    Article  CAS  PubMed  Google Scholar 

  48. Bramson JL et al. Direct intratumoral injection of an adenovirus expressing interleukin-12 induces regression and long-lasting immunity that is associated with highly localized expression of interleukin-12 Hum Gene Ther 1996 7: 1995–2002

    Article  CAS  PubMed  Google Scholar 

  49. Rakhmilevich AL et al. Cytokine gene therapy of cancer using gene gun technology: superior antitumor activity of interleukin-12 Hum Gene Ther 1997 8: 1303–1311

    Article  CAS  PubMed  Google Scholar 

  50. Coor M, Lee DJ, Carson DA, Tighe H . Gene vaccination with naked plasmid DNA: mechanism of CTL priming J Exp Med 1996 184: 1555–1560

    Article  Google Scholar 

  51. Ulmer JB et al. Generation of MHC class I-restricted cytotoxic T lymphocytes by expression of a viral protein in muscle cells: antigen presentation by non-muscle cells Immunol 1996 89: 59–67

    Article  CAS  Google Scholar 

  52. Iwasaki A et al. The dominant role of bone marrow-derived cells in CTL induction following plasmid DNA immunization at different sites J Immunol 1997 159: 11–14

    CAS  PubMed  Google Scholar 

  53. Conry RM et al. Selected strategies to augment polynucleotide immunization Gene Therapy 1996 3: 67–74

    CAS  PubMed  Google Scholar 

  54. Behrens L et al. Human muscle cells express a functional costimulatory molecule distinct from B7.1 (CD80) and B7.2 (CD86) in vitro and in inflammatory lesions J Immunol 1998 161: 5943–5951

    CAS  PubMed  Google Scholar 

  55. Erwing LP, Kluth DC, Walsh GM, Ress AJ . Initial cytokine exposure determines function of macrophages and renders them corresponsive to other cytokines J Immunol 1998 161: 1983–1988

    Google Scholar 

  56. Sato Y et al. Immune-stimulatory DNA sequences necessary for effective intradermal gene immunization Science 1996 273: 352–354

    Article  CAS  PubMed  Google Scholar 

  57. Tepper RI, Coffman RL, Leder P . An eosinophil-dependent mechanism for the anti-tumor effect of interleukin-4 Science 1992 257: 548–551

    Article  CAS  PubMed  Google Scholar 

  58. Chomarat P, Rybak ME, Banchereau J . Interleukin-4. In: Thomson A (ed) The Cytokine Handbook Academic Press: San Diego 1998 pp 133–174

    Google Scholar 

  59. I. Xiang Z, Ertl HC . Manipulation of the immune response to a plasmid-encoded viral antigen by coinoculation with plasmids expressing cytokines Immunity 1995 2: 129–135

    Article  PubMed  Google Scholar 

  60. Hartikka J et al. An improved plasmid DNA expression vector for direct injection into skeletal muscle Hum Gene Ther 1996 7: 1205–1217

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Dr Abraham Fuks (McGill University, Montreal, Canada) and Dr Serge Jothy (Sunnybrook Hospital, Toronto, Ontario, Canada) for kindly providing reagents and cell lines, Dr Fawaz Halwani (McGill University) for assistance with FACS analyses, and Dr Fu Hu (Montreal General Hospital, Montreal, Canada) for assistance with statistical analyses. This study was funded by the Fraser Fund (Royal Victoria Hospital, Montreal, Canada). Keli Song was supported in part by Royal Victoria Hospital Research Institute Fellowship.

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  1. Department of Pathology, McGill University, 3775 University Street, Room 13, Montreal, H3A 2B4, Quebec, Canada

    K Song, Y Chang & G J Prud'homme

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Song, K., Chang, Y. & Prud'homme, G. Regulation of T-helper-1 versus T-helper-2 activity and enhancement of tumor immunity by combined DNA-based vaccination and nonviral cytokine gene transfer. Gene Ther 7, 481–492 (2000). https://doi.org/10.1038/sj.gt.3301123

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