Nina King - Academia.edu (original) (raw)
Papers by Nina King
Molecular Nutrition & Food Research, 2005
Attempts to treat peanut allergy using traditional methods of allergen desensitization are accomp... more Attempts to treat peanut allergy using traditional methods of allergen desensitization are accompanied by a high risk of anaphylaxis. The aim of this study was to determine if modifications to the IgE-binding epitopes of a major peanut allergen would result in a safer immunotherapeutic agent for the treatment of peanut-allergic patients. IgE-binding epitopes on the Ara h 2 allergen were modified, and modified Ara h 2 (mAra h 2) protein was produced. Wild-type (wAra h 2) and mAra h 2 proteins were analyzed for their ability to interact with T-cells, their ability to bind IgE, and their ability to release mediators from a passively sensitized RBL-2H3 cell line. Multiple T-cell epitopes were identified on the major peanut allergen, Ara h 2. Ara h 2 amino acid regions 11-35, 86-125, and 121-155 contained the majority of peptides that interact with T-cells from most patients. The wAra h 2 and mAra h 2 proteins stimulated proliferation of T-cells from peanut-allergic patients to similar levels. In contrast, the mAra h 2 protein exhibited greatly reduced IgE-binding capacity compared to the wild-type allergen. In addition, the modified allergen released significantly lower amounts of beta-hexosaminidase, a marker for IgE-mediated RBL-2H3 degranulation, compared to the wild-type allergen.
Journal of Allergy and Clinical Immunology, 2002
International Archives of Allergy and Immunology, 2001
Background: Numerous strategies have been proposed for the treatment of peanut allergies, but des... more Background: Numerous strategies have been proposed for the treatment of peanut allergies, but despite the steady advancement in our understanding of atopic immune responses and the increasing number of deaths each year from peanut anaphylaxis, there is still no safe, effective, specific therapy for the peanut-sensitive individual. Immunotherapy would be safer and more effective if the allergens could be altered to reduce their ability to initiate an allergic reaction without altering their ability to desensitize the allergic patient. Methods: The cDNA clones for three major peanut allergens, Ara h 1, Ara h 2, and Ara h 3, have been cloned and characterized. The IgE-binding epitopes of each of these allergens have been determined and amino acids critical to each epitope identified. Site-directed mutagenesis of the allergen cDNA clones, followed by recombinant production of the modified allergen, provided the reagents necessary to test our hypothesis that hypoallergenic proteins are e...
International Archives of Allergy and Immunology, 1999
Allergy to peanut is a significant IgE-mediated health problem because of the high prevalence, po... more Allergy to peanut is a significant IgE-mediated health problem because of the high prevalence, potential severity, and chronicity of the reaction. Currently the only treatment for peanut hypersensitivity is avoidance of the food which is inadequate as shown by the fact that 50% of peanut-sensitive individuals in a recent study [1] reported a significant accidental reaction within a 2-year period. Therefore, the goal of our laboratory is to develop specific, safe, and effective immunotherapy for patients suffering from peanut hypersensitivity. A potentially safe and efficacious immunotherapeutic strategy would be one that utilized allergenic proteins modified in such a way so they no longer bind IgE but still could bind T cells and either elicit a Th 1 type response or lead to tolerance. Ara h 2, an abundant peanut protein, is recognized by serum IgE from c90% of peanut-sensitive individuals [2]. It has been shown to belong to the conglutin family of seed storage proteins and contain 10-linear IgE-binding epitopes three of which were immunodominant [3]. Our previous work has shown that individual epitopes, when taken out of the context of the intact allergen, could be mutated to non-IgE-binding sites by changing a single amino acid to alanine [3]. We wanted to take this concept one step further and produce a gene that had the immunodominant epitopes changed to non-IgEbinding sites, produce this modified version of the allergen Isolation and Characterization of Allergens
Archives of Biochemistry and Biophysics, 1997
proved diagnostic and therapeutic approaches to peanut hypersensitivity. ᭧ 1997 Academic Press A ... more proved diagnostic and therapeutic approaches to peanut hypersensitivity. ᭧ 1997 Academic Press A major peanut allergen, Ara h 2, is recognized by serum IgE from ú90% of patients with peanut hypersensitivity. Biochemical characterization of this allergen indicates that it is a glycoprotein of Ç17.5 kDa. Using N-terminal amino acid sequence data from puri
Molecular Nutrition & Food Research, 2005
Attempts to treat peanut allergy using traditional methods of allergen desensitization are accomp... more Attempts to treat peanut allergy using traditional methods of allergen desensitization are accompanied by a high risk of anaphylaxis. The aim of this study was to determine if modifications to the IgE-binding epitopes of a major peanut allergen would result in a safer immunotherapeutic agent for the treatment of peanut-allergic patients. IgE-binding epitopes on the Ara h 2 allergen were modified, and modified Ara h 2 (mAra h 2) protein was produced. Wild-type (wAra h 2) and mAra h 2 proteins were analyzed for their ability to interact with T-cells, their ability to bind IgE, and their ability to release mediators from a passively sensitized RBL-2H3 cell line. Multiple T-cell epitopes were identified on the major peanut allergen, Ara h 2. Ara h 2 amino acid regions 11-35, 86-125, and 121-155 contained the majority of peptides that interact with T-cells from most patients. The wAra h 2 and mAra h 2 proteins stimulated proliferation of T-cells from peanut-allergic patients to similar levels. In contrast, the mAra h 2 protein exhibited greatly reduced IgE-binding capacity compared to the wild-type allergen. In addition, the modified allergen released significantly lower amounts of beta-hexosaminidase, a marker for IgE-mediated RBL-2H3 degranulation, compared to the wild-type allergen.
Journal of Allergy and Clinical Immunology, 2002
International Archives of Allergy and Immunology, 2001
Background: Numerous strategies have been proposed for the treatment of peanut allergies, but des... more Background: Numerous strategies have been proposed for the treatment of peanut allergies, but despite the steady advancement in our understanding of atopic immune responses and the increasing number of deaths each year from peanut anaphylaxis, there is still no safe, effective, specific therapy for the peanut-sensitive individual. Immunotherapy would be safer and more effective if the allergens could be altered to reduce their ability to initiate an allergic reaction without altering their ability to desensitize the allergic patient. Methods: The cDNA clones for three major peanut allergens, Ara h 1, Ara h 2, and Ara h 3, have been cloned and characterized. The IgE-binding epitopes of each of these allergens have been determined and amino acids critical to each epitope identified. Site-directed mutagenesis of the allergen cDNA clones, followed by recombinant production of the modified allergen, provided the reagents necessary to test our hypothesis that hypoallergenic proteins are e...
International Archives of Allergy and Immunology, 1999
Allergy to peanut is a significant IgE-mediated health problem because of the high prevalence, po... more Allergy to peanut is a significant IgE-mediated health problem because of the high prevalence, potential severity, and chronicity of the reaction. Currently the only treatment for peanut hypersensitivity is avoidance of the food which is inadequate as shown by the fact that 50% of peanut-sensitive individuals in a recent study [1] reported a significant accidental reaction within a 2-year period. Therefore, the goal of our laboratory is to develop specific, safe, and effective immunotherapy for patients suffering from peanut hypersensitivity. A potentially safe and efficacious immunotherapeutic strategy would be one that utilized allergenic proteins modified in such a way so they no longer bind IgE but still could bind T cells and either elicit a Th 1 type response or lead to tolerance. Ara h 2, an abundant peanut protein, is recognized by serum IgE from c90% of peanut-sensitive individuals [2]. It has been shown to belong to the conglutin family of seed storage proteins and contain 10-linear IgE-binding epitopes three of which were immunodominant [3]. Our previous work has shown that individual epitopes, when taken out of the context of the intact allergen, could be mutated to non-IgE-binding sites by changing a single amino acid to alanine [3]. We wanted to take this concept one step further and produce a gene that had the immunodominant epitopes changed to non-IgEbinding sites, produce this modified version of the allergen Isolation and Characterization of Allergens
Archives of Biochemistry and Biophysics, 1997
proved diagnostic and therapeutic approaches to peanut hypersensitivity. ᭧ 1997 Academic Press A ... more proved diagnostic and therapeutic approaches to peanut hypersensitivity. ᭧ 1997 Academic Press A major peanut allergen, Ara h 2, is recognized by serum IgE from ú90% of patients with peanut hypersensitivity. Biochemical characterization of this allergen indicates that it is a glycoprotein of Ç17.5 kDa. Using N-terminal amino acid sequence data from puri