OBJECTIVE:
To 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.
METHODS:
Based 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 β-lactoglobulin.
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.
RESULTS:
The 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.
CONCLUSIONS:
The 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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