Beruflich Dokumente
Kultur Dokumente
spread over most areas of Japan in the early spring. C. japonica pollen causes cedar
pollinosis with rhinitis, asthma and conjunctivitis as clinical symptoms. About 20% of
the Japanese population is currently afflicted, and more than half of the Japanese
population has specific circulating immunoglobulin E (IgE) for cedar pollen allergens.
Two major allergens, designated Cry j 1 and Cry j 2, have been isolated from
Japanese cedar pollen and characterized in detail. More than 90% of cedar pollinosis
Allergen-specific immunotherapy is the only treatment that can provide a cure for
side-effects such as anaphylactic shock due to the presence of IgE-binding activity, and
pain caused by inflammation. A safe, easy and convenient treatment would thus be a
Peptide immunotherapy using dominant T-cell epitopes has been shown to be a safe
and effective treatment for the control of IgE-mediated allergic diseases because of the
have demonstrated that two rice-based edible vaccines expressing either mouse T-cell
epitopes or seven-linked human dominant T-cell epitopes (7Crp), derived from Cry j 1
peptide expressed in rice seed, and indicate that rice seed-based peptide vaccines can be
However, T cell epitopes differ from each other dependent upon the genotypes
peptide immunotherapy.
Furthermore, the clinical use of a rice seed-based edible vaccine with low IgE binding
activity for humans and other mammals affected by cedar pollinosis requires the
plants that specifically express the entire T cell epitope Cry j 1 peptide in seeds. We
alternative to T-cell epitope peptides. Our preliminary results showed that partial- or
full-length Cry j 1 peptide were barely accumulated in the endosperm of transgenic rice
seed even under the control of the strong rice endosperm-specific GluB-1 promoter.
protein with rice glutelin. Three overlapping fragments covering the entire Cry j 1
region were inserted into the highly variable C terminal region of the GluA-2 acidic
subunit in the GluA-2 pro-glutelin precursor (Fig. 1). The highest accumulation level of
the fusion protein reached about 15% of total seed protein, but fusion protein precursors
containing Cry j 1 with an altered structure were not post-translationally processed into
mature forms and thus aggregated with Cys-rich prolamins in protein body I (PB-I) of
seed endosperm tissues.
Transgenic plants have been used as attractive bioreactors for the production of
plant systems offer several advantages over conventional competing systems such as
microbial and mammalian cell culture systems with regard to lower production cost,
easy control of production scale and low risk of contamination of mammalian pathogens.
Choosing the ideal host plant or tissue for expression of recombinant proteins is an
important factor. We investigated the tissue and intracellular localization suitable for
production of artificial recombinant 7Crp peptide. This artificial peptide could be only
amounts of transcripts in vegetative tissues such as leaf and stem, when it was expressed
under the control of constitutive promoters such as rice AGPase large subunit and
maizeubiquitin-1 promoters (Fig. 2). These results indicate that endosperm tissue is the
transgenic rice.
BamH1 BglII HindIII Nco 1Sac1 EcoR 1
pAg7 hpt CaMV35S P GluB -1 P SP GluB -1 T
LB RB
1 353
pJ1full Cry j1 full KDEL
1 195
pJ1N -half Cry j1 N -half KDEL
135 353
pJ1C -half Cry j1 C -half KDEL
1 144
Cry j1 F1 Cry j1 F1
126 257
Cry j1 F2 Cry j1 F2
231 353
Cry j1 F3
Cry j1 F3 Con V1-F1 V2-F1 V2-F2 V2-F3
1 144
pV2-F1 Cry j1 F1
126 257
pV2-F2 Cry j1 F2
231 353
pV2-F3 Cry j1 F3
S L Sm S L Sm S L Sm S L Sm S L Sm S L Sm
75 k
50 k
37 k
25 k
20 k
15 k
10 k