Disruption of allergenic activity of the major grass pollen allergen Phl p 2 by reassembly as a mosaic protein.
The recognition of conformational epitopes on respiratory allergens by IgE Abs is a key event in allergic inflammation. We report a molecular strategy for the conversion of allergens into vaccines with reduced allergenic activity, which is based on the reassembly of non-IgE-reactive fragments in the form of mosaic proteins. This evolution process is exemplified for timothy grass pollen-derived Phl p 2, a major allergen for more than 200 million allergic patients. In a first step, the allergen was disrupted into peptide fragments lacking IgE reactivity. cDNAs coding for these peptides were reassembled in altered order and expressed as a recombinant mosaic molecule. The mosaic molecule had lost the three-dimensional structure, the IgE reactivity, and allergenic activity of the wild-type allergen, but it induced high levels of allergen-specific IgG Abs upon immunization. These IgG Abs crossreacted with group 2 allergens from other grass species and inhibited allergic patients' IgE binding to the wild-type allergen. The mosaic strategy is a general strategy for the reduction of allergenic activity of protein allergens and can be used to convert harmful allergens into safe vaccines.
Mothes-Luksch
Nadine
N
Division of Immunopathology, Department of Pathophysiology, Center of Physiology, Pathophysiology and Immunology, Vienna General Hospital, Medical University of Vienna, Vienna, Austria.
Stumvoll
Sabine
S
Linhart
Birgit
B
Focke
Margit
M
Krauth
Marie-Therese
MT
Hauswirth
Alexander
A
Valent
Peter
P
Verdino
Petra
P
Pavkov
Tea
T
Keller
Walter
W
Grote
Monika
M
Valenta
Rudolf
R
D000485:Allergens / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003888:Desensitization, Immunologic / Q000379:methods; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D006109:Poaceae / Q000235:genetics* / Q000276:immunology; D011058:Pollen / Q000235:genetics* / Q000276:immunology; D015202:Protein Engineering; D011817:Rabbits; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology / Q000517:prevention & control; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism
D000485:Allergens; D007074:Immunoglobulin G; C084984:PHLPII protein, Phleum pratense; D010940:Plant Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D007073:Immunoglobulin E