Affiliations: [a] MRC Collaborative Centre, 1-3 Burtonhole Lane, Mill Hill, London UK | [b] Athena Neurosciences, 800F Gateway Boulevard, South San Francisco, CA USA
Correspondence:
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Correspondence and reprint requests to: Dr. Olivier Léger, MRC Collaborative Centre, 1-3 Burtonhole Lane, Mill Hill, London, NW7 1AD, UK.
Abstract: α4β1 integrin (VLA-4) is crucialfor the adhesion of leukocytes to human vascular cell adhesion molecule-1 (VCAM-1) on iriflamed endothelium. This cell adhesion event is the first step in leukocyte extravasation across the blood-brain barrier in iriflammatory diseases of the central nervous system (CNS) such as experimental autoimmune encephalomyelitis (EAE). Prevention of leukocyte infiltration by antibodies against the α4 integrin, which block the α4β1 integrin/VCAM-1 interaction, have been shown to suppress clinical and pathological features of EAE. In this study, two mouse monoclonal antibodies (MAb) directed against human α4 integrin were analyzed in vitro for their ability to block the interaction of leukocytes with VCAM-1 under different assay conditions. The best blocking MAb, AN100226m, was humanized by complementarily-determining region grafting, associated with human C regions and expressed. We found that modification of two structural determinants (H27 and H29) for the heavy chain CDRI loop in one hand, and modification of framework amino acid H38, H40 and H44 in the other hand, had no effect on antigen binding. In contrast, modification of a structural determinant (H71) for the heavy chain CDR2 loop resulted in loss of binding. The humanized antibody, AN100226, was equivalent to the murine antibody, AN100226m, in binding to α4β1 integrin and in blocking cell adhesion. More importantly, AN100226 was as effective as AN100226m in the reversal of active EAE in guinea pigs and thus may be useful in the treatment of autoimmune diseases such as multiple sclerosis. AN100226 is currently in phase II clinical trials in the UK for the treatment of multiple sclerosis exacerbations.
