In search of a tolerance-induction strategy for cow's milk allergies: significant reduction of beta-lactoglobulin allergenicity via transglutaminase/cysteine polymerization.
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 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.


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.
Laboratory of Immunology and Experimental Allergy, Department of Internal Medicine, Faculty of Medical Sciences, University of Campinas, Campinas/SP, Brazil.
Celso Eduardo
CE
Olivier
Regiane Patussi dos Santos
RP
Lima
Daiana Guedes
DG
Pinto
Raquel Acacia Pereira Goncalves dos
RA
Santos
Grayce Katlen Moreno da
GK
Silva
Sônia Letícia Silva
SL
Lorena
Mariana Battaglin
MB
Villas-Boas
Flávia Maria
FM
Netto
Ricardo de Lima
Rde L
Zollner
D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000276:immunology; D016022:Case-Control Studies; D003545:Cysteine / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006361:Heating; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology*; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000097:blood; D007782:Lactoglobulins / Q000276:immunology*; D007957:Leukocyte Adherence Inhibition Test; D008297:Male; D008875:Middle Aged; D016269:Milk Hypersensitivity / Q000276:immunology* / Q000517:prevention & control; D058105:Polymerization; D012882:Skin Tests; D018709:Statistics, Nonparametric; D011503:Transglutaminases / Q000737:chemistry / Q000276:immunology*; D055815:Young Adult
D000485:Allergens; D007782:Lactoglobulins; D007073:Immunoglobulin E; D011503:Transglutaminases; D003545:Cysteine