Effect of heat treatment and enzymatic digestion on the B cell epitopes of cow's milk proteins.
BACKGROUND
Processing milk leads to changes in clinical allergenicity. However, the mechanism by which heat treatment affects the allergenicity of milk proteins is not fully understood.


OBJECTIVE
We investigated the effect of heat treatment and enzymatic digestion on the allergenicity of B cell epitopes of milk proteins using a histamine release assay.


METHODS
Human basophils were passively sensitized using sera from 10 patients with allergies to cow's milk. All the patients experienced symptoms immediately after ingesting milk. The human basophils were obtained from umbilical cord blood mononuclear cells after culturing the mononuclear cells for 3-4 weeks in the presence of IL-3. After sensitization with 10% patient sera for 48 h, the cells were stimulated with untreated, heat-treated, or heat-treated and pepsin-and-trypsin-digested beta-lactoglobulin or alpha-casein for 1 h. The histamine concentrations in the supernatants were then measured by radioimmunoassay.


RESULTS
Heat treatment alone did not alter the molecular weight of beta-lactoglobulin or alpha-casein. Heat treatment of beta-lactoglobulin significantly increased its susceptibility to enzymatic digestion in a time- and temperature-dependent manner and reduced its ability to induce histamine release from sensitized basophils. Similar findings were not observed for alpha-casein. The combination of heat treatment and enzymatic digestion reduced the abilities of both beta-lactoglobulin and alpha-casein to induce histamine release from passively sensitized basophils.


CONCLUSIONS
Heat treatment reduced the allergenicity of beta-lactoglobulin by inducing conformational changes and by increasing its susceptibility to enzymatic digestion, both of which disrupted B cell epitopes, whereas heat treatment alone did not alter the allergenicity of alpha-casein.
Morisawa
Y
Y
Department of Pediatrics, Kochi Medical School, Kochi, Japan.
Kitamura
A
A
Ujihara
T
T
Zushi
N
N
Kuzume
K
K
Shimanouchi
Y
Y
Tamura
S
S
Wakiguchi
H
H
Saito
H
H
Matsumoto
K
K
D000293:Adolescent; D000485:Allergens / Q000737:chemistry / Q000276:immunology; D000818:Animals; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D002364:Caseins / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006632:Histamine / Q000276:immunology; D006636:Histamine Release / Q000276:immunology; D006358:Hot Temperature; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007223:Infant; D007231:Infant, Newborn; D007782:Lactoglobulins / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008892:Milk / Q000276:immunology*; D016269:Milk Hypersensitivity / Q000276:immunology*; D010434:Pepsin A / Q000737:chemistry; D011487:Protein Conformation; D014357:Trypsin / Q000737:chemistry
D000485:Allergens; D002364:Caseins; D018985:Epitopes, B-Lymphocyte; D007782:Lactoglobulins; D007073:Immunoglobulin E; D006632:Histamine; D014357:Trypsin; D010434:Pepsin A