BACKGROUND: Food allergies are more prevalent in children, due to the immature gastrointestinal epithelial membrane barrier allowing more proteins through the barrier and into circulation. Ovomucoid (OM) is one of the major allergens that is found in egg white. OBJECTIVE: The aim of this study was to determine T cell epitopes, antigen-presenting human leucocyte antigen (HLA) class II molecules of the T cell lines (TCLs) and T cell clones (TCCs), and complementarity determining region (CDR) 3 loops of the T cell receptor (TCR) alpha and beta chains of the TCCs specific to OM. METHODS: We established TCLs and TCCs specific to OM from peripheral blood mononuclear cells (PBMCs) of four atopic patients with egg-white allergy using a mixture of a panel of overlapping synthetic peptides corresponding to the amino acid sequence of the entire OM. We identified the T cell epitopes by antigen-induced proliferative responses, antigen-presenting molecules using allogeneic PBMCs and CDR3 loops of the TCR alpha and beta chains by cloning and sequence analysis. RESULTS: The TCLs and TCCs responded to seven different peptides, and their antigen-presenting molecules were different from each other. Sequence analysis of the TCR alpha and beta gene usage of the TCCs showed marked heterogeneity, and the usage of the CDR3 loop of the TCCs involved heterogenous amino acid residues. Interestingly, TCCs 'IH3.3' and 'YT6.1' recognized the same OM peptides, and had the same TCR Vbeta-Jbeta gene usage. Considering that peptide motifs bind to HLA class II molecules, the electrically charged residue (positive or negative) on the CDR3alpha and the CDR3beta loops of TCR of TCC may form ionic bonds with a charged residue on the HLA class II molecules-peptide complex. CONCLUSIONS: TCCs that have the same TCR gene usage were established from patients who had shown similar hypersensitivity-type, indicating that antigen recognition by a specific TCR is closely associated with the characteristics of each patient's symptoms.
BACKGROUND:Food allergies are more prevalent in children, due to the immature gastrointestinal epithelial membrane barrier allowing more proteins through the barrier and into circulation. Ovomucoid (OM) is one of the major allergens that is found in egg white. OBJECTIVE: The aim of this study was to determine T cell epitopes, antigen-presenting human leucocyte antigen (HLA) class II molecules of the T cell lines (TCLs) and T cell clones (TCCs), and complementarity determining region (CDR) 3 loops of the T cell receptor (TCR) alpha and beta chains of the TCCs specific to OM. METHODS: We established TCLs and TCCs specific to OM from peripheral blood mononuclear cells (PBMCs) of four atopic patients with egg-white allergy using a mixture of a panel of overlapping synthetic peptides corresponding to the amino acid sequence of the entire OM. We identified the T cell epitopes by antigen-induced proliferative responses, antigen-presenting molecules using allogeneic PBMCs and CDR3 loops of the TCR alpha and beta chains by cloning and sequence analysis. RESULTS: The TCLs and TCCs responded to seven different peptides, and their antigen-presenting molecules were different from each other. Sequence analysis of the TCR alpha and beta gene usage of the TCCs showed marked heterogeneity, and the usage of the CDR3 loop of the TCCs involved heterogenous amino acid residues. Interestingly, TCCs 'IH3.3' and 'YT6.1' recognized the same OM peptides, and had the same TCR Vbeta-Jbeta gene usage. Considering that peptide motifs bind to HLA class II molecules, the electrically charged residue (positive or negative) on the CDR3alpha and the CDR3beta loops of TCR of TCC may form ionic bonds with a charged residue on the HLA class II molecules-peptide complex. CONCLUSIONS: TCCs that have the same TCR gene usage were established from patients who had shown similar hypersensitivity-type, indicating that antigen recognition by a specific TCR is closely associated with the characteristics of each patient's symptoms.