RESUMO
Japanese cedar pollen allergen Cry j2 is a causal allergen of seasonal pollinosis in Japan. To analyze B cell epitopes of Cry j2, we established two human-mouse hybridomas secreting IgM class human monoclonal antibodies to Cry j2. A pin-peptide enzyme-linked immunosorbent assay with synthesized icosa peptides showed that 404-117 monoclonal antibody bound to peptides #11-13 with cry j2 amino acid sequence of 101F-L140. Detailed analysis with octa peptides and alanine substituted peptides indicated that an amino acid sequence of 118FKVD121 was an essential for antibody binding. When K119 (Asn) was substituted with alanine, 404-117 monoclonal antibody did not bind to the alanine substituted peptide. We concluded that the 118FKVD121 sequence might have a very important role in early recognition by Cry j2-specific B cells, which could act as antigen presenting cells.
Assuntos
Anticorpos Monoclonais/biossíntese , Epitopos/imunologia , Imunoglobulina M/biossíntese , Proteínas de Plantas/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Alérgenos/química , Alérgenos/imunologia , Sequência de Aminoácidos , Animais , Especificidade de Anticorpos , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/patologia , Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Linfócitos B/imunologia , Linfócitos B/patologia , Sítios de Ligação , Cryptomeria/química , Cryptomeria/imunologia , Epitopos/química , Humanos , Hibridomas/imunologia , Hibridomas/metabolismo , Japão , Camundongos , Peptídeos/química , Peptídeos/imunologia , Proteínas de Plantas/química , Pólen/química , Ligação Proteica , Rinite Alérgica Sazonal/induzido quimicamente , Rinite Alérgica Sazonal/patologiaRESUMO
We obtained a stable human-mouse hybridoma clone 4701-1 secreting IgM class human monoclonal antibody to Japanese cedar pollen allergen Cry j1. A pin-peptide enzyme-linked immunosorbent assay (ELISA) with synthesized pentadeca peptides showed a peptide with an amino acid sequence of LYTVT NSDDD PVNPA was found to be positive. Detailed analysis with deca to tetra peptides indicated that an amino acid sequence of TVTN was an essential sequence for antibody binding. When N (Asn) was substituted with A (Ala) of the TVTN epitope, the resulting peptide did not have antibody binding ability. We concluded that the TVTN sequence might have a very important role in early recognition of Cry j1 allergen by Cry j1-specific B cells, which act as antigen presenting cells.