A neutralizing anti-Nogo66 receptor monoclonal antibody reverses inhibition of neurite outgrowth by central nervous system myelin.
The Nogo66 receptor (NgR1) is a neuronal, leucine-rich repeat (LRR) protein that binds three central nervous system (CNS) myelin proteins, Nogo, myelin-associated glycoprotein, and oligodendrocyte myelin glycoprotein, and mediates their inhibitory effects on neurite growth. Although the LRR domains on NgR1 are necessary for binding to the myelin proteins, the exact epitope(s) involved in ligand binding is unclear. Here we report the generation and detailed characterization of an anti-NgR1 monoclonal antibody, 7E11. The 7E11 monoclonal antibody blocks Nogo, myelin-associated glycoprotein, and oligodendrocyte myelin glycoprotein binding to NgR1 with IC50 values of 120, 14, and 4.5 nm, respectively, and effectively promotes neurite outgrowth of P3 rat dorsal root ganglia neurons cultured on a CNS myelin substrate. Further, we have defined the molecular epitope of 7E11 to be DNAQLR located in the third LRR domain of rat NgR1. Our data demonstrate that anti-NgR1 antibodies recognizing this epitope, such as 7E11, can neutralize CNS myelin-dependent inhibition of neurite outgrowth. Thus, specific anti-NgR1 antibodies may represent a useful therapeutic approach for promoting CNS repair after injury.
Li
Weiwei
W
Biogen Idec, Inc, Cambridge, Massachusetts 02142, USA.
Walus
Lee
L
Rabacchi
Sylvia A
SA
Jirik
Adrienna
A
Chang
Ernie
E
Schauer
Jessica
J
Zheng
Betty H
BH
Benedetti
Nancy J
NJ
Liu
Betty P
BP
Choi
Eugene
E
Worley
Dane
D
Silvian
Laura
L
Mo
Wenjun
W
Mullen
Colleen
C
Yang
Weixing
W
Strittmatter
Stephen M
SM
Sah
Dinah W Y
DW
Pepinsky
Blake
B
Lee
Daniel H S
DH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000918:Antibody Specificity; D001923:Brain Chemistry; D002417:Cattle; D000939:Epitopes / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D005727:Ganglia, Spinal / Q000187:drug effects / Q000502:physiology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000276:immunology*; D009186:Myelin Sheath / Q000187:drug effects / Q000502:physiology*; D016501:Neurites / Q000187:drug effects / Q000502:physiology*; D000070798:Nogo Proteins; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009185:Myelin Proteins; D000070798:Nogo Proteins; D010446:Peptide Fragments; C000603199:RTN4 protein, human; D011994:Recombinant Proteins; C000603201:Rtn4 protein, mouse; C000603197:Rtn4 protein, rat