Induction of antibodies to particular sites of the alpha7 subunit of the nicotinic acetylcholine receptor in mice of different lines

• Published in: Bioorganicheskaia khimiia Vol. 33; no. 4; p. 442
• Main Authors: Koroev, D O, Titova, M A, Volkova, T D, Oboznaia, M B, Filatova, M P, Fufacheva, E N, Zhmak, M N, Tsetlin, V I, Bobkova, N V, Vol'pina, O M
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01.07.2007
• Subjects: alpha7 Nicotinic Acetylcholine Receptor; Amino Acid Sequence; Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - blood; Antibody Affinity; Humans; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Molecular Sequence Data; Peptide Fragments - immunology; Rats; Receptors, Nicotinic - immunology
• ISSN: 0132-3423

Five synthetic fragments of the N-terminal domain of the alpha7 subunit of the human nicotinic acetylcholine receptor (alpha7 nAChR) that correspond to theoretically calculated B epitopes and T helper epitopes of the protein and contain from 16 to 29 amino acid residues were tested for the ability to stimulate the formation of antibodies in mice of three lines having H-2d, H-2b, and H-2k haplotypes of the major histocompatibility complex. It was shown that, in the free (unconjugated) form, all the peptides stimulate the formation of antibodies at least in one mouse line. Most of the peptides induced the formation of antibodies in BALB/c mice (haplotype H-2d); therefore, more detailed studies were carried out on these animals. The free peptides and/or their conjugates with keyhole limpet hemocyanin were demonstrated to be capable of stimulating the formation in BALB/c mice of antibodies that bind to the recombinant extracellular N-terminal domain of (alpha7 nAChRalpha). The epitope mapping of antipeptide antibodies carried out using truncated fragments helped reveal antipeptide antibodies to four regions of the alpha7 subunit: 1-23, 98-106, 159-168, and 173-188 (or 179-188).