Depression of T-cell epitope generation by stabilizing hen lysozyme.
Conformational stability of proteins is an important factor that determines their resistance/susceptibility to proteolytic digestion. Intracellular proteolysis is the key step in antigen presentation events for protein antigens; hence, it is likely that increasing protein stability reduces the antigenicity of proteins. We prepared three hen egg white lysozyme derivatives possessing different stabilities by chemical modification to clarify the relationship between conformational stability and the antigenicity of the protein. One of the derivatives was conformationally unstabilized by removing one intramolecular disulfide bond, whereas the two others were stabilized by the addition of an intramolecular cross-link. The antigenicity of these derivatives was evaluated using hen egg white lysozyme-specific T-cell hybridoma cells and a B-lymphoma cell line, A20, as antigen-presenting cells. With an increase in conformational stability, the T-cell response decreased. However, the reduction was not derived from the inefficiency of internalization to A20 cells nor the alteration of antigenicity by chemical modifications. Moreover, from analyses of their susceptibility to proteolysis and the kinetics of presentation of the T-cell epitope, it was confirmed that increasing protein stability led to the depression of T-cell epitope generation by increasing resistance to proteolysis. These results have an important implication in devising a new strategy for manipulating T-cell response by the stability of protein antigen.
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Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-82, Japan.
Ito
H O
HO
Koga
T
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Watanabe
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Ueda
T
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Imoto
T
T
D000818:Animals; D000941:Antigens / Q000378:metabolism; D002645:Chickens; D004705:Endocytosis; D018984:Epitopes, T-Lymphocyte / Q000096:biosynthesis*; D006868:Hydrolysis; D007700:Kinetics; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D014407:Tumor Cells, Cultured
D000941:Antigens; D018984:Epitopes, T-Lymphocyte; D009113:Muramidase