Return to Contents in this Issue

Original Article


Production and Immunogenicity of Hypoallergenic Codon-Optimized DNA Vaccine Encoding Mature Der p 1 Allergen


P Pulsawat,1 S Piboonpocanun,2 S Sirivichayakul,1 S Buranapraditkun,1 A Jacquet,1 M Shimada,3 K Okuda,3 K Ruxrungtham1

1Chulalongkorn University, Bangkok, Thailand
2Institute of Molecular Biosciences, Mahidol University, Nakhon pathom, Thailand
3Yokohama City University, Yokohama, Japan

J Investig Allergol Clin Immunol 2010; Vol. 20(7): 582-590



Background: Genetic vaccination with plasmid DNA encoding allergens is a promising potential approach for the treatment or prevention of allergy. Nonetheless, because the allergens expressed can display immunoglobulin (Ig) E reactivity, methods to deliver hypoallergenicvariants can minimize the risk of type 2 helper (TH2) cell priming after DNA immunization.

Methods: A humanized synthetic gene encoding mature Dermatophagoides pteronyssinus group 1 (Der p 1) allergen was cloned into the pHIS expression vector carrying unmethylated CpG 2006 (CpG 2006) motif but devoid of signal sequence. The immunogenicity of this DNA construct was compared in naïve mice with that of recombinant ProDer p 1 protein adjuvanted with alum.

Results: Codon optimization of the cDNA encoding mature Der p 1 markedly improved allergen expression. Mature Der p 1, expressed intracellularly in Human Embryonic Kidney 293 cells (HEK 293 cells) transfected with codon-optimized Der p 1 cDNA (pHIS-mHuDer p 1), was shown to be hypoallergenic as it displayed no IgE reactivity. Intradermal vaccinations of naïve Balb/C mice with pHIS-mHuDer p 1 elicited an allergen-specific TH1 response characterized by the production of specifi c IgG2a, a very low amount of specific IgG1, and no specific IgE. Lipoplex formulation with cationic liposome composed of lecithin, N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methylsulfate (DOTAP) and cholesterol not only accelerated the induction of TH1 response but also increased its intensity.

Conclusion: A codon-optimized DNA vaccine encoding mature Der p 1 in a lipoplex formulation could represent a promising hypoallergenic vaccine candidate for safer immunotherapy against house dust mite allergy.

Key words: Hypoallergen. Der p 1. Codon-optimized DNA vaccine. Mature Der p 1.