TY - JOUR
T1 - The cleavage site of C5 from man and animals as a common target for neutralizing human monoclonal antibodies
T2 - In vitro and in vivo studies
AU - Marzari, Roberto
AU - Sblattero, Daniele
AU - Macor, Paolo
AU - Fischetti, Fabio
AU - Gennaro, Renato
AU - Marks, James D.
AU - Bradbury, Andrew
AU - Tedesco, Francesco
PY - 2002/10
Y1 - 2002/10
N2 - The isolation of an anti-C5 single-chain fragment variable (scFv) antibody, TS-A12/22, from a human phage display library, is described. This antibody inhibits the activation of C5 and the assembly of the terminal complement complex implicated in cell and tissue damage. Using antibody-sensitized sheep erythrocytes and rabbit red cells as target cells in hemolytic assays, we found that TS-A12/22 inhibited the activation of C5 by the convertases of both classical and alternative pathways. Western blot analysis and competition experiments with synthetic peptides showed that TS-A12/22 reacted with the α chain of C5 and recognized the cleavage site of this complement component by the C5 convertase. As a result, the antibody prevented splitting of C5 and inhibited the generation of C5a and of the terminal complement complex. The identification of the TS-A12/22 recognition site as a conserved sequence in man, mouse, rat and rabbit enabled the demonstration of in vitro inhibition of complement activity in these species. The scFv TS-A12/22 was tested in a rat model of antigen-induced arthritis and proved to be effective in preventing influx of polymorphonuclear cells into the knee joint and C9 deposition on synovial tissue.
AB - The isolation of an anti-C5 single-chain fragment variable (scFv) antibody, TS-A12/22, from a human phage display library, is described. This antibody inhibits the activation of C5 and the assembly of the terminal complement complex implicated in cell and tissue damage. Using antibody-sensitized sheep erythrocytes and rabbit red cells as target cells in hemolytic assays, we found that TS-A12/22 inhibited the activation of C5 by the convertases of both classical and alternative pathways. Western blot analysis and competition experiments with synthetic peptides showed that TS-A12/22 reacted with the α chain of C5 and recognized the cleavage site of this complement component by the C5 convertase. As a result, the antibody prevented splitting of C5 and inhibited the generation of C5a and of the terminal complement complex. The identification of the TS-A12/22 recognition site as a conserved sequence in man, mouse, rat and rabbit enabled the demonstration of in vitro inhibition of complement activity in these species. The scFv TS-A12/22 was tested in a rat model of antigen-induced arthritis and proved to be effective in preventing influx of polymorphonuclear cells into the knee joint and C9 deposition on synovial tissue.
KW - Complement
KW - Inflammation
KW - Phage display library
KW - Single-chain fragment variable antibody
UR - http://www.scopus.com/inward/record.url?scp=0036772528&partnerID=8YFLogxK
U2 - 10.1002/1521-4141(2002010)32:10<2773::AID-IMMU2773>3.0.CO;2-G
DO - 10.1002/1521-4141(2002010)32:10<2773::AID-IMMU2773>3.0.CO;2-G
M3 - Article
SN - 0014-2980
VL - 32
SP - 2773
EP - 2782
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 10
ER -