Production of an agonist-like monoclonal antibody to the human A2A receptor of adenosine for clinical use.
The second extracellular loop of the A(2A) receptor (A(2A)R) of adenosine was used to immunize mice for production of Adonis, an IgM monoclonal antibody. Adonis bound to the immunogen peptide and the native receptor in ELISA with K(D) values in 6.51-12.35 nM range. It recognized a linear epitope of 7 amino acids (LFEDVVP) at the C-terminal part of the external loop. Adonis revealed a 45-kDa band in lysate of human peripheral blood mononuclear cells in Western blotting in denaturing conditions. This served to monitor the up-regulation of the A(2A)R expression by caffeine. Adonis stimulated the cAMP production and inhibited the cell proliferation of an A(2A)R transfected stable cell line. These results confirm the immunogenicity and the functional relevance of the second extracellular loop of the A(2A)R. They suggest that Adonis may be of clinical use in various pathological situations to measure the regulation of the A(2A)R expression and to act as A(2A)R agonist drug.
By
Youlet
Y
Laboratoire de Biochimie Endocrinienne et Métabolique (EA3288), Université de la Méditerranée, Faculté de Médecine Timone, Marseille, France.
Durand-Gorde
Josée-Martine
JM
Condo
Jocelyne
J
Lejeune
Pierre-Jean
PJ
Mallet
Bernard
B
Carayon
Pierre
P
Guieu
Régis
R
Ruf
Jean
J
D058908:Adenosine A2 Receptor Agonists; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D000242:Cyclic AMP / Q000096:biosynthesis; D018604:Epitope Mapping; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D043705:Receptor, Adenosine A2A / Q000276:immunology*; D017186:Titrimetry
D058908:Adenosine A2 Receptor Agonists; D000911:Antibodies, Monoclonal; D010455:Peptides; D043705:Receptor, Adenosine A2A; D000242:Cyclic AMP