Background: DNA immunization and protein immunization with CpG motifs as adjuvants represent promising approaches in allergen-specific immunotherapy. Objective: We investigated the effect of coinjection or prepriming with CpG-ODN on Th2-type responses induced by gene gun and protein immunization. Methods: BALB/c mice were immunized with the gene gun using plasmid DNA containing the cDNAs coding for the genes of Bet v 1a, Phl p 2 and β-galactosidase or with the purified Al(OH)3-adsorbed proteins. In addition, CpG-ODN were applied by coinjection or by prepriming treatment. Antibody and cytokine responses were measured by ELISA, proliferative and cytotoxic responses were determined by standard labeling procedures. Furthermore, the allergenic activity of sera was measured by passive cutaneous anaphylaxis. Results: Gene gun immunization and protein immunization induced a clear Th2-type response for all antigens. The Th1-promoting effect of CpG-ODN coinjection together with gene gun immunization was restricted to β-galactosidase as indicated by the increase of IgG2a and a marked expression of IFN-γ. CpG motifs also increased the specific cytotoxic response against β-galactosidase. Prepriming with CpG-ODN and gene gun or protein immunization with Bet v 1a exhibited no significant difference to the non-CpG control group. However, sera from mice preprimed with CpG-ODN induced no anaphylaxis with gene gun immunization, but with protein immunization. Conclusions: The effect of CpG motifs in vivo depends on a variety of parameters like the nature of the antigen and the immunization modality. Furthermore, our studies indicate that a combination of CpG + DNA immunization may be more effective in antagonizing Th2 responses than the combination of CpG + protein immunization.

Keywords:
DNA immunization, DNA immunotherapeutics, CpG motifs, BALB/c, Bet v 1, Phl p 2, β-Galactosidase
1.
Donnelly JJ, Ulmer JB, Shiver JW, Liu MA: DNA vaccines. Annu Rev Immunol 1997;15:617–648.
2.
Stevenson FK: DNA vaccines against cancer: From genes to therapy. Ann Oncol 1999;10:1413–1418.
3.
Hsu CH, Chua KY, Tao MH, Lai YL, Wu HD, Huang SK, Hsieh KH: Immunoprophylaxis of allergen-induced immunoglobulin E synthesis and airway hyperresponsiveness in vivo by genetic immunization. Nat Med 1996;2:540–544.
4.
Hartl H, Kiesslich J, Weiss R, Bernhaupt A, Mostbock S, Scheiblhofer S, Ebner C, Ferreira F, Thalhamer J: Immune responses after immunization with plasmid DNA encoding Bet v 1, the major allergen of birch pollen. J Allergy Clin Immunol 1999;103:107–113.
5.
Raz E, Tighe H, Sato Y, Corr M, Dudler JA, Roman M, Swain SL, Spiegelberg HL, Carson DA: Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization. Proc Natl Acad Sci USA 1996;93:5141–5145.
6.
Krieg AM, Yi AK, Matson S, Waldschmidt TJ, Bishop GA, Teasdale R, Koretzky GA, Klinman DM: CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 1995;374:546–549.
7.
Klinman DM, Yi AK, Beaucage SL, Conover J, Krieg AM: CpG motifs present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12 and interferon gamma. Proc Natl Acad Sci USA 1996;93:2879–2883.
8.
Kobayashi H, Horner AA, Takabayashi K, Nguyen MD, Huang E, Cinman N, Raz E: Immunostimulatory DNA prepriming: A novel approach for prolonged Th1-biased immunity. Cell Immunol 1999;198:69–75.
9.
Feltquate DM, Heaney S, Webster RG, Robinson HL: Different T helper cell types and antibody isotypes generated by saline and gene gun DNA immunization. J Immunol 1997;158:2278–2284.
10.
Brtko J, Mostbock S, Scheiblhofer S, Hartl A, Thalhamer J: DNA immunization is associated with increased activity of type I iodothyronine 5′-deiodinase in mouse liver. Mol Cell Endocrinol 1999;152:85–89.
11.
Hartl A, Kiesslich J, Weiss R, Bernhaupt A, Mostbock S, Scheiblhofer S, Flockner H, Sippl M, Ebner C, Ferreira F, Thalhamer J: Isoforms of the major allergen of birch pollen induce different immune responses after genetic immunization. Int Arch Allergy Immunol 1999;120:17–29.
12.
Abrams SI, Stanziale SF, Lunin SD, Zaremba S, Schlom J: Identification of overlapping epitopes in mutant ras oncogene peptides that activate CD4+ and CD8+ T cell responses. Eur J Immunol 1996;26:435–443.
13.
Faquim-Mauro EL, Coffman RL, Abrahamsohn IA, Macedo MS: Cutting edge: Mouse IgG1 antibodies comprise two functionally distinct types that are differentially regulated by IL-4 and IL-12. J Immunol 1999;163:3572–3576.
14.
Breiteneder H, Pettenburger K, Bito A, Valenta R, Kraft D, Rumpold H, Scheiner O, Breitenbach M: The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene. EMBO J 1989;8:1935–1938.
15.
Dolecek C, Vrtala S, Laffer S, Steinberger P, Kraft D, Scheiner O, Valenta R: Molecular characterization of Phl p II, a major timothy grass (Phleum pratense) pollen allergen. FEBS Lett 1993;335:299–304.
16.
Kline JN, Waldschmidt TJ, Businga TR, Lemish JE, Weinstock JV, Thorne PS, Krieg AM: Cutting edge: Modulation of airway inflammation by CpG oligodeoxynucleotides in a murine model of asthma. J Immunol 1998;160:2555–2559.
17.
Lee SW, Sung YC: Immuno-stimulatory effects of bacterial-derived plasmids depend on the nature of the antigen in intramuscular DNA inoculations. Immunology 1998;94:285–289.
© 2001 S. Karger AG, Basel
2001
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.