• Celso Eduardo Olivier, Regiane Patussi dos Santos Lima, Daiana Guedes Pinto, Raquel Acacia Pereira Goncalves dos Santos, Grayce Katlen Moreno da Silva, Sônia Letícia Silva Lorena, Mariana Battaglin Villas-Boas, Flávia Maria Netto, and Ricardo de Lima Zollner.
• Laboratory of Immunology and Experimental Allergy, Department of Internal Medicine, Faculty of Medical Sciences, University of Campinas, Campinas/SP, Brazil.
• Clinics (Sao Paulo). 2012 Oct 1; 67 (10): 117111791171-9.

ObjectiveTo explore the use of β-lactoglobulin polymerized using microbial transglutaminase and heating to identify whether protein polymerization could reduce in vivo allergenicity and maintain in vitro and ex vivo immunoreactivity for use in tolerance-induction protocols.MethodsBased on previous protocols applied in mice and children, we performed in vivo challenges (using a skin prick test) with native and polymerized β-lactoglobulin in adult patients with an IgE-mediated allergy to plactoglobulin. In vitro humoral immunoreactivity was analyzed using immunoblotting. Cell-mediated immunoreactivity was analyzed using ex vivo challenges with native and polymerized β-lactoglobulin and monitored by leukocyte adherence inhibition tests.ResultsThe skin tests demonstrated that there was a significant reduction in immediate cutaneous reactivity after polymerization. Polymerization did not decrease the immunoblotting detection of s-IgE specific to β-lactoglobulin. Cell-mediated immunoreactivity, as assessed by ex vivo challenges and leukocyte adherence inhibition tests, did not exhibit significant differences between leukocytes challenged with native versus polymerized β-lactoglobulin.ConclusionsThe polymerization of β-lactoglobulin decreased in vivo allergenicity and did not decrease in vitro humoral or ex vivo cell-mediated immunoreactivity. Therefore, we conclude that inducing polymerization using transglutaminase represents a promising technique to produce suitable molecules for the purpose of designing oral/ sublingual tolerance induction protocols for the treatment of allergies.

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