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1.
Clin Exp Allergy ; 36(5): 666-75, 2006 May.
Article in English | MEDLINE | ID: mdl-16650053

ABSTRACT

BACKGROUND: In contrast to other Rosaceae fruit, only few cases of patients with adverse reactions to strawberry are listed in literature. OBJECTIVE To identify allergenic proteins in strawberry and to express and characterize recombinant strawberry lipid transfer protein (LTP; rFra a 3). METHODS: Established apple-allergic patients were recruited on the basis of a reported allergic reaction to strawberry (n=28, confirmed by double-blind placebo-controlled food challenge in four patients) or on the basis of IgE reactivity to LTP (n=34). Sensitization to purified natural and recombinant allergens was assessed by RAST, immunoblot (inhibition) and basophil histamine release (BHR). A strawberry cDNA library was screened for genes homologous to known fruit allergens. Fra a 3 was cloned and expressed in the yeast Pichia pastoris and compared with peach and apple LTP by RAST, immunoblot-inhibition and BHR tests. RESULTS: Genes homologous to Bet v 1, Bet v 6, profilin and LTP were identified in a strawberry cDNA library. In BHR the rFra a 3 induced histamine release at a 100-fold higher concentration than peach LTP. RAST inhibition showed high cross-reactivity to peach and apple LTP, although IgE reactivity was lower by a factor 5. On strawberry immunoblot, patients' IgE showed reactivity to a Bet v 1 homologue, profilin, LTP and high-molecular weight bands. CONCLUSION: In addition to a Bet v 1 homologue, strawberry also contains IgE-binding profilin and LTP. The rFra a 3 has less allergenic potency than peach and apple LTP, and therefore is an interesting tool for future immunotherapy. Fra a 3 does not seem to be clinically relevant.


Subject(s)
Carrier Proteins/immunology , Food Hypersensitivity/immunology , Fragaria/immunology , Plant Proteins/immunology , Profilins/immunology , Allergens/genetics , Allergens/immunology , Antigens, Plant/genetics , Antigens, Plant/immunology , Basophils/immunology , Carrier Proteins/genetics , Cohort Studies , Cross Reactions/immunology , Double-Blind Method , Histamine Release/immunology , Humans , Immunoblotting/methods , Immunoglobulin E/immunology , Italy , Plant Proteins/genetics , Radioallergosorbent Test/methods , Recombinant Proteins/immunology , Spain
2.
Dev Biol (Basel) ; 121: 143-50, 2005.
Article in English | MEDLINE | ID: mdl-15962477

ABSTRACT

Different vaccination methods have been applied to protect fish against the detrimental effects of various pathogens. Several studies have shown the potentials of oral vaccination. In theory oral vaccination is an effortless and stress-free method which can be applied at almost any age. In general, however, the vaccine has to be protected to avoid digestion, which results in high costs for application in aquaculture. In this paper we introduce a cost-effective oral vaccination strategy for viral diseases of fish. The vaccines discussed here include fusion proteins consisting of a gut adhesion molecule and a viral peptide expressed in plants. The adhesion molecule mediates binding to and uptake from the gut, whereas the viral peptide functions as vaccine antigen mediating the induction of a humoral immune response. The first pilot studies using a fusion of the gut adhesion molecule and well-characterised heterologous linear B- and T-cell viral epitopes, produced in potato tubers, showed a promising binding and subsequent uptake in the end gut of carp. The results further indicated that a specific humoral immune response was evoked.


Subject(s)
Aquaculture/methods , Carps , Fish Diseases/prevention & control , Vaccination/methods , Vaccination/veterinary , Viral Vaccines/administration & dosage , Virus Diseases/veterinary , Administration, Oral , Animals , Bacterial Toxins/metabolism , Blotting, Western/veterinary , Enterotoxins/metabolism , Enzyme-Linked Immunosorbent Assay/veterinary , Epitopes, T-Lymphocyte/metabolism , Escherichia coli Proteins/metabolism , Histological Techniques/veterinary , Immunohistochemistry/veterinary , Solanum tuberosum/metabolism , Solanum tuberosum/virology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/metabolism , Viral Vaccines/metabolism , Virus Diseases/prevention & control
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