Investigation of the Binding and Outside-In Signalling Mechanisms of Anti-CD11d Acute Neurotrauma Therapy
Abstract
Neurotrauma is a debilitating injury, and no treatment currently exists that improves both physical and neurological recovery. A novel immunomodulatory therapeutic agent – anti-CD11d – has been developed at Robarts Research Institute. Application of anti-CD11d therapy following injury decreases the infiltration of peripheral leukocytes into the injured central nervous system, improves physical recovery, and improves neurological recovery in rodent neurotrauma models. The binding and signalling mechanisms of anti-CD11d on leukocytes, however, remain poorly understood. An endogenous CD11d/CD18 expression model was established in differentiated THP-1 cells to investigate anti-CD11d mechanisms. Immunocytochemistry and Förster Resonance Energy Transfer (FRET) microscopy characterized anti-CD11d binding to active and inactive CD11d/CD18 conformations. Western blot analysis did not detect CD11d/CD18 signalling through tyrosine kinases following stimulation with low or moderate concentrations of soluble anti-CD11d (CB1/CA1). Overall, my study will inform the development of anti-CD11d acute neurotrauma therapy and provide new tools to investigate basic CD11d/CD18 biology.