Preparation and characterization of agonistic monoclonal antibodies against Toll-like receptor 4-MD-2 complex.
Ligands for toll-like receptors (TLR) are known to induce a variety of immune responses. Selective induction of desirable responses would be important for the treatment of individual diseases with various pathogenesis. For this purpose, we established six MAbs against the TLR4/MD-2 complex (UT MAbs) from TLR4(-/-) mice or MD-2(-/-) mice. Three MAbs (UT12, 18, and 22) induced NF-kappaB activation and production of pro-inflammatory cytokines, but the other three (UT15, 41, and 49) did not induce such cell responses. Unlike lipopolysaccharide (LPS), agonistic UT MAbs did not require serum components for the functions. UT41 and UT49 recognized TLR4 in the absence of MD-2. On the other hand, the other four MAbs reacted to the TLR4/MD-2 complex, but not to solo TLR4. Agonistic UT MAbs shared the epitopes, but non-agonistic UT15 reacted to distinct epitope on the complex. UT MAbs appear to be useful analyzing the molecular mechanism of TLR signaling and will contribute to the development of novel immunotherapies.
Bahrun
Uleng
U
Department of Immunology, Saga Medical School, Saga, Japan.
Kimoto
Masao
M
Tsukamoto
Hiroki
H
Tsuneyoshi
Naoko
N
Kohara
Jun
J
Fukudome
Kenji
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002460:Cell Line; D006801:Humans; D006825:Hybridomas; D051218:Lymphocyte Antigen 96 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016513:Mice, SCID; D011485:Protein Binding; D051197:Toll-Like Receptor 4 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; C495377:LY96 protein, human; C411728:Ly96 protein, mouse; D051218:Lymphocyte Antigen 96; C495345:TLR4 protein, human; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4