Abstract
To optimize polynucleotide vaccinations for protective antitumor immunity we used a self-replicating RNA vaccine in which Semliki Forest virus replicase drives RNA expression of the lacZ gene coding for β-galactosidase as model tumor-associated antigen (TAA). This was compared with replicase-deficient control RNA and with lacZ DNA plasmids with respect to gene expression in vitro and in vivo and for vaccination using the mouse ear pinna as an optimal immunization site. In vitro, the highest expression was observed with self-replicating RNA. Gene expression following pinna inoculation of either non-replicating DNA plasmids or self-replicating RNA was similar, lasting for 2–3 weeks. Higher antibody responses were obtained with RNA than with DNA. β-Gal peptide specific CTL memory responses to lacZ DNA or RNA lasted for more than 6 weeks while respective responses induced by lacZ-transfected tumor cells lasted for only 2 weeks. To achieve a protective response against lacZ tumor cells with self-replicating RNA about a 100-fold lower dose of polynucleotide was sufficient in comparison to DNA. The extent of protective antitumor immunity not only depended on the gene dose used for vaccination, but also on the aggressiveness of the lacZ- transfected tumor line used for challenge. In comparison to lacZ-transfected tumor cells as vaccines, polynucleotide vaccination also demonstrated superiority with regard to cross-protection. Protective antitumor immunity could be strongly increased upon co-inoculation of lacZ DNA with IL-2 DNA or IL-12 RNA. IL-2 DNA, but not IL-12 RNA, also augmented the CTL response while IL-12 RNA, but not IL-2 DNA, reduced the antibody response. These results demonstrate efficient protective antitumor immunity after intra-pinna lacZ TAA polynucleotide vaccination and show additional immunomodulatory effects by co-administration of cytokine polynucleotides.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Donnelly JJ, Ulmer JB, Shiver JW, Liu MA . DNA vaccines Annu Rev Immunol 1997 15: 617–648
Barry MA, Johnston SA . Biological features of genetic immunization Vaccine 1997 8: 788–791
Hargest R, Williamson R . Prophylactic gene therapy for cancer Gene Therapy 1996 3: 97–102
Moelling K . Naked DNA – the poor man's gene therapy? Editorial Gene Therapy 1998 5: 573–574
Whalen RG . DNA vaccines for infectious diseases, allergies and cancer. In: Eibl et al (eds) Symposium in Immunology VII 1998 pp 165–183
Förg P, von Hoegen P, Dalemans W, Schirrmacher V . Superiority of the ear pinna over muscle tissue as site for DNA vaccination Gene Therapy 1998 5: 789–797
Liljeström, P, Garoff H . A new generation of animal cell expression vectors based on the Semliki Forest virus replicon Biotechnology 1991 9: 1356–1361
Meanger J, Peroulis I, Mills J . Modified Semliki Forest virus expression vector that facilitates cloning BioTechniques 1997 23: 432–436
Carbone FR, Bevan J . Class I-restricted progressing and presentation of exogenous cell-associated antigen in vivo J Exp Med 1990 171: 377–387
Khazaie K et al. Persistence of dormant tumor cells in the bone marrow of tumor cell vaccinated mice correlates with long-term immunological protection Proc Natl Acad Sci USA 1994 91: 7430–7434
Krüger A, Schirrmacher V, von Hoegen P . Scattered micrometastasis visualized at the single cell level: detection and re-isolation of lacZ labeled metastasized lymphoma cells Int J Cancer 1994 58: 275–284
Krüger A el al. Pattern and load of spontaneous liver metastasis dependent on host immune status studied with a lacZ-transduced lymphoma Blood 1994 84: 3166–3174
Raz E . Gene Vaccination: Theory and Practice Springer: Berlin 1998 pp 1–180
Conry RM, LoBuglio A, Curiel DT . Polynucleotide-mediated immunization therapy of cancer Semin Oncol 1996 23: 135–147
Dubensky TW Jr, Polo JM, Liu MA . Live virus vaccines: something old, something new, something borrowed Nature Med 1998 4: 1357–1358
Conry RM et al. Characterization of a messenger RNA polynucleotide vaccine vector Cancer Res 1995 55: 1397–1400
Qiu P, Ziegelhoffer P, Sun J, Yang NS . Gene gun delivery of mRNA in situ results in efficient transgene expression and genetic immunization Gene Therapy 1996 3: 262–268
Hahn CS, Hahn YS, Braciale TJ, Rice CM . Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation Proc Natl Acad Sci USA 1992 89: 2679–2683
Mandl CW et al. In vitro synthesized infections RNA as an attenuated live vaccine in a flavivirus model Nature Med 1989 4: 1438–1440
Ying H et al. Cancer therapy using a self-replicating RNA vaccine Nature Med 1999 5: 823–827
Leitner WW et al. Enhancement of tumor-specific immune response with plasmid DNA replicon vectors Cancer Res 2000 60: 51–55
Leitner WW, Ying H, Restifo NP . DNA and RNA-based vaccines: principles, progress and prospects Vaccine 2000 18: 765–777
Condon C et al. DNA-based immunization by in vivo transfection of dendritic cells Nature Med 1996 2: 1122–1128
Grohmann U et al. Dendritic cells, interleukin 12 and CD4+ lymphocytes in the initiation of class I-restricted reactivity to a tumor/self peptide Crit Rev Immunol 1998 18: 87–98
Casares S et al. Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility class II-restricted viral epitope J Exp Med 1997 186: 1481–1486
Progador A et al. Predominant role for directly transfected dendritic cells in antigen presentation to CD8+ T cells after gene gun immunization J Exp Med 1998 188: 1075–1082
Akbari O et al. DNA vaccination: transfection and activation of dendritic cells as key events for immunity J Exp Med 1999 189: 169–177
Lynch DH, Namen AE, Miller RE . In vivo evaluation of the effects of interleukins 2, 4 and 7 on enhancing the immunotherapeutic efficacy of anti-tumor cytotoxic T lymphocytes Eur J Immunol 1991 21: 2977–2985
Sparwasser T et al. Bacterial DNA and immuno-stimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells Eur J Immunol 1998 28: 2045–2054
Boccaccio GL, Mor F, Steinmann L . Non-coding plasmid DNA induces IFN-γ in vivo and suppresses autoimmune encephalomyelitis Int Immunol 1999 11: 289–296
Kovacsovics-Bankowski M, Clark K, Benacerraf B, Rock KL . Efficient major histocompatibility complex I presentation of exogenous antigen upon phagocytosis by macrophages Proc Natl Acad Sci USA 1993 90: 4942–4944
Bachmann MF et al. TAP1-independent loading of class I molecules by exogenous viral proteins Eur J Immunol 1995 25: 1739–1743
Albert ML, Sauter B, Bhardwaj N . Dendritic cells acquire antigen from apoptotic cells and induce class I restricted CTLs Nature 1998 392: 86–89
Lau LL, Jamieson BD, Somasundaran T, Ahmed R . Cytotoxic T cell memory without antigen Nature 1994 369: 648–652
Opferman IT, Ober BT, Ashton-Rickardt PG . Linear differentiation of cytotoxic effectors into memory T lymphocytes Science 1999 283: 1745–1748
Klinman DM et al. Contribution of cells at the site of DNA vaccination to the generation of antigen-specific immunity and memory J Immunol 1998 160: 2388–2392
Sigal LJ, Crotty S, Andino R, Rock KL . Cytotoxic T cell immunity to virus-infected non-haematopoietic cells requires presentation of exogenous antigen Nature 1999 398: 77–80
Schirrmacher V, Zangemeister-Wittke U . γ-Irradiation suppresses T cell mediated protective immunity against a metastatic tumor in the afferent phase of the immune response but enhances it in the efferent phase if given before immune cell transfer Int J Oncol 1994 4: 335–346
Schild HJ, Kyewski B, von Hoegen P, Schirrmacher V . CD4+ helper T cells are required for resistance to a highly metastatic murine tumor Eur J Immunol 1987 17: 1863–1866
Jurianz K, von Hoegen P, Schirrmacher V . Superiority of the ear pinna over a subcutaneous tumor inoculation site for induction of a TH1-type cytokine response Cancer Immunol Immunother 1998 45: 327–333
Schirrmacher V, Jurianz K, Griesbach A . Intra-pinna induction of specific antitumor immune T cell functions: effect of ear resection after antigen application Int J Oncol 1997 11: 227–233
Ridge JP, Di Rosa F, Matzinger P . A conditioned dendritic cell can be a temporal bridge between a CD4 T-helper and a T-killer cell Nature 1998 393: 474–478
Ramarathinam L et al. Multiple lineages of tumors express a common tumor antigen, P1A, but they are not cross-protected J Immunol 1995 155: 5323–5329
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
Irvine KR, Rao JB, Rosenberg SA, Restifo NP . Cytokine enhancement of DNA immunization leads to effective treatment of established pulmonary metastases J Immunol 1996 156: 238–245
Zhu N, Liggit D, Liu Y, Debs R . Systemic gene expression after intravenous DNA delivery into adult mice Science 1993 261: 209–211
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
Waisman A et al. Suppressive vaccination with DNA encoding a variable region gene of the T cell receptor prevents autoimmune encephalitis and activates Th2 immunity Nature Med 1996 2: 899–905
Prayaga SK, Ford MJ, Haynes JR . Manipulation of HIV-1 gp120-specific immune responses elicited via gene gun-based DNA immunization Vaccine 1997 15: 1349–1352
Von Hoegen P . Gene tag: just a way to follow tumor cells or induction of new antigens Cancer J 1995 8: 31
Abram SI, Hand PH, Tsang KY, Schlom J . Mutant ras epitopes as targets for cancer vaccines Semin Oncol 1996 23: 118–134
Theobald M et al. Targeting p53 as a general tumor antigen Proc Natl Acad Sci USA 1995 92: 11993–11997
Coulie P et al. A mutated intron sequence for an antigenic peptide recognized by cytolytic T lymphocytes on a human melanoma Proc Natl Acad Sci USA 1995 92: 7976–7980
Ciernik FI, Berzofsky JA, Carbone DP . Induction of cytotoxic T lymphocytes and antitumor immunity with DNA vaccine expressing single T cell epitopes J Immunol 1996 156: 2369–2375
Chen CH et al. Gene gun-mediated DNA vaccination induces antitumor immunity against human papillomavirus type 16 E7-expressing murine tumor metastases in the liver and lungs Gene Therapy 1999 6: 1972–1981
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schirrmacher, V., Förg, P., Dalemans, W. et al. Intra-pinna anti-tumor vaccination with self-replicating infectious RNA or with DNA encoding a model tumor antigen and a cytokine. Gene Ther 7, 1137–1147 (2000). https://doi.org/10.1038/sj.gt.3301220
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301220
Keywords
This article is cited by
-
N1-methylpseudouridine-incorporated mRNA enhances exogenous protein expression and suppresses immunogenicity in primary human fibroblast-like synoviocytes
Cytotechnology (2022)
-
Viromers as carriers for mRNA-mediated expression of therapeutic molecules under inflammatory conditions
Scientific Reports (2020)
-
Optimization of CFTR-mRNA transfection in human nasal epithelial cells
Translational Medicine Communications (2016)
-
mRNA-based therapeutics — developing a new class of drugs
Nature Reviews Drug Discovery (2014)
-
Vaccination with mRNAs encoding tumor-associated antigens and granulocyte-macrophage colony-stimulating factor efficiently primes CTL responses, but is insufficient to overcome tolerance to a model tumor/self antigen
Cancer Immunology, Immunotherapy (2006)