A hypoallergenic vaccine obtained by tail-to-head restructuring of timothy grass pollen profilin, Phl p 12, for the treatment of cross-sensitization to profilin.
Profilins are highly cross-reactive allergens in pollens and plant food. In a paradigmatic approach, the cDNA coding for timothy grass pollen profilin, Phl p 12, was used as a template to develop a new strategy for engineering an allergy vaccine with low IgE reactivity. Non-IgE-reactive fragments of Phl p 12 were identified by synthetic peptide chemistry and restructured (rs) as a new molecule, Phl p 12-rs. It comprised the C terminus of Phl p 12 at its N terminus and the Phl p 12 N terminus at its C terminus. Phl p 12-rs was expressed in Escherichia coli and purified to homogeneity. Determination of secondary structure by circular dichroism indicated that the restructuring process had reduced the IgE-reactive alpha-helical contents of the protein but retained its beta-sheet conformation. Phl p 12-rs exhibited reduced IgE binding capacity and allergenic activity but preserved T cell reactivity in allergic patients. IgG Abs induced by immunization of mice and rabbits with Phl p 12-rs cross-reacted with pollen and food-derived profilins. Recombinant Phl p 12-rs, rPhl p 12-rs, induced less reaginic IgE to the wild-type allergen than rPhl p 12. However, the rPhl p 12-rs-induced IgGs inhibited allergic patients' IgE Ab binding to profilins to a similar degree as those induced by immunization with the wild type. Phl p 12-rs specific IgG inhibited profilin-induced basophil degranulation. In conclusion, a restructured recombinant vaccine was developed for the treatment of profilin-allergic patients. The strategy of tail-to-head reassembly of hypoallergenic allergen fragments within one molecule represents a generally applicable strategy for the generation of allergy vaccines.
Westritschnig
Kerstin
K
Christian Doppler Laboratory for Allergy Research, Division of Immunopathology, Department of Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
Linhart
Birgit
B
Focke-Tejkl
Margarete
M
Pavkov
Tea
T
Keller
Walter
W
Ball
Tanja
T
Mari
Adriano
A
Hartl
Arnulf
A
Stöcklinger
Angelika
A
Scheiblhofer
Sandra
S
Thalhamer
Josef
J
Ferreira
Fatima
F
Vieths
Stefan
S
Vogel
Lothar
L
Böhm
Alexandra
A
Valent
Peter
P
Valenta
Rudolf
R
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018926:Anti-Allergic Agents / Q000737:chemistry / Q000276:immunology*; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology; D000937:Antigen-Antibody Reactions; D052179:Antigens, Plant / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001665:Binding Sites; D002942:Circular Dichroism; D000939:Epitopes / Q000276:immunology; D005818:Genetic Engineering / Q000379:methods; D006632:Histamine / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008958:Models, Molecular; D011058:Pollen / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D051304:Profilins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology; D014612:Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000485:Allergens; D018926:Anti-Allergic Agents; D000906:Antibodies; D052179:Antigens, Plant; D000939:Epitopes; D007074:Immunoglobulin G; C526193:Phl p 12 allergen, timothy grass; D051304:Profilins; D011993:Recombinant Fusion Proteins; D014612:Vaccines; D007073:Immunoglobulin E; D006632:Histamine