Downstream effect profiles discern different mechanisms of integrin αLβ2 inhibition.
The integrin leucocyte function-associated antigen-1 (αLβ2, LFA-1) plays crucial roles in T cell adhesion, migration and immunological synapse (IS) formation. Consequently, αLβ2 is an important therapeutic target in autoimmunity. Three major classes of αLβ2 inhibitors with distinct modes of action have been described to date: Monoclonal antibodies (mAbs), small molecule α/β I allosteric and small molecule α I allosteric inhibitors. The objective of this study was to systematically compare these three modes of αLβ2 inhibition for their αLβ2 inhibitory as well as their potential agonist-like effects. All inhibitors assessed were found to potently block αLβ2-mediated leucocyte adhesion. None of the inhibitors induced ZAP70 phosphorylation, indicating absence of agonistic outside-in signalling. Paradoxically, however, the α/β I allosteric inhibitor XVA143 induced conformational changes within αLβ2 characteristic for an intermediate affinity state. This effect was not observed with the α I allosteric inhibitor LFA878 or the anti-αLβ2 mAb efalizumab. On the other hand, efalizumab triggered the unscheduled internalization of αLβ2 in CD4+ and CD8+ T cells while LFA878 and XVA143 did not affect or only mildly reduced αLβ2 surface expression, respectively. Moreover, efalizumab, in contrast to the small molecule inhibitors, disturbed the fine-tuned internalization/recycling of engaged TCR/CD3, concomitantly decreasing ZAP70 expression levels. In conclusion, different modes of αLβ2 inhibition are associated with fundamentally different biologic effect profiles. The differential established here is expected to provide important translational guidance as novel αLβ2 inhibitors will be advanced from bench to bedside.
Division of Clinical Pharmacology and Toxicology and Department of Research, University Hospital, CH-4031 Basel, Switzerland.
Riccardo V
RV
Mancuso
Novartis Pharma AG, Novartis Institutes of Biomedical Research, CH-4002 Basel, Switzerland.
Karl
K
Welzenbach
Institute of Immunology, Medical University of Vienna, Lazarettgasse 19, 1090 Vienna, Austria.
Peter
P
Steinberger
Division of Clinical Pharmacology and Toxicology and Department of Research, University Hospital, CH-4031 Basel, Switzerland.
Stephan
S
Krähenbühl
AlloCyte Pharmaceuticals AG, Hochbergerstrasse 60C, CH-4057 Basel, Switzerland. Electronic address: gabriele.weitz-schmidt@unibas.ch.
Gabriele
G
Weitz-Schmidt
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized; D002448:Cell Adhesion; D002460:Cell Line; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D016023:Integrins / Q000819:agonists / Q000037:antagonists & inhibitors*; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism*; D051379:Mice
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D016023:Integrins; D016169:Lymphocyte Function-Associated Antigen-1; C472181:efalizumab
Allosteric inhibitor; Integrin; LFA-1; Paradoxical agonism; TCR