Characterization of porcine CD19 and anti-CD19 monoclonal antibodies.
CD19 is an important pan B cell marker and co-stimulatory protein in humans and mice. Efforts to further characterize B cell ontogeny in swine have been hampered by the lack of monoclonal antibodies (mAb) to valuable surface markers like Vpre-B, CD19, CD34 and CD43. We report here on the complete nucleotide and deduced amino acid sequence of porcine CD19, the cross-reactivity of anti-human CD19 monoclonals and efforts to prepare anti-porcine CD19 mAb to bacterially-expressed products. Porcine CD19 is highly homologous to those in the few other species studied, i.e. human, mouse and guinea pig, but only in certain domains. Among the 14 CD19 exons, homology approaches 90% to human CD19 in exons 6, 9, 11 and 12 and is approximately 80% with other species in this region. The highly homologous C-terminal cytoplasmic region contains nine tyrosines including the YEND/E motif that binds the SH2 domain of Fyn. Two different porcine CD19 isoforms that differ in their 3' UTRs were identified just as in human CD19. Thus, the signaling properties of CD19 may be similar to those in humans. On the other hand, only 60% sequence similarity was seen in exons 1-5 that encode the N-terminal extracellullar region that is involved in ligand binding and is the target of CD19-specific mAb. This probably explains why only 1 of the 17 anti-human CD19 mAb tested recognized swine B cells. Furthermore, when the extracellular domains of CD19 were expressed in E. coli, mAbs to the bacterially-expressed product did not recognize CD19 on porcine B cells suggesting that carbohydrate-dependent conformation may determine antigenicity.
Sun
Jishan
J
Department of Microbiology, The University of Iowa, Iowa City, IA 52242-1109, USA.
Sinkora
Jiri
J
Wertz
Nancy
N
Moravkova
Alena
A
Butler
J E
JE
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D018941:Antigens, CD19 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D006801:Humans; D008969:Molecular Sequence Data; D020033:Protein Isoforms; D017385:Sequence Homology; D013045:Species Specificity; D013552:Swine
D000911:Antibodies, Monoclonal; D018941:Antigens, CD19; D020033:Protein Isoforms