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Importantly, and in contrast to ILT4, NRP1 interacted with CSPs that were covalently bound to target surfaces in the course of complement activation, therefore representing a classical complement receptor. We demonstrate that binding of C4d to NRP1 expressing cells was dose-dependent and saturable, and had a K D value of 0.71 μM.

Although NRP1 contains domains homologous to ones found in some complement proteins, it has not been linked to the complement system. NRP1 is also expressed on immune cells and serves as a marker for murine Tregs. NRP1, a highly conserved type 1 transmembrane protein, plays important roles in the development of the nervous and cardiovascular system as well as in tumorigenesis through interaction with its established binding partners, such as vascular endothelial growth factor (VEGF) and semaphorin 3A (Sema3A). By applying an unbiased screening approach using a cDNA mammalian expression library generated from human monocyte-derived dendritic cells and probed with recombinant human C4d, we identified neuropilin-1 (NRP1) as a novel receptor for C4d, C3d, and iC3b. Since immunoglobulin-like transcript 4 (ILT4) was previously shown to interact with soluble CSPs, but not with CSPs covalently-bound to target surfaces following classical complement activation, the present study aimed to identify novel cellular receptors interacting with covalently-deposited CSPs.

5Division of Clinical and Experimental Immunology, Center for Pathophysiology, Infectiology, and Immunology, Institute of Immunology, Medical University of Vienna, Vienna, AustriaĬomplement split products (CSPs), such as the fragments C4d and C3d, which are generated as a consequence of complement regulatory processes, are established markers for disease activity in autoimmunity or antibody-mediated graft rejection.

4Division of Nephrology and Dialysis, Department of Internal Medicine III, Medical University Vienna, Vienna, Austria.

3Departments of Biochemistry and Immunology, University of Toronto, Toronto, ON, Canada.

2Department of Clinical Cell Biology and FACS Core Unit, Children's Cancer Research Institute (CCRI), Vienna, Austria.

1Division of Immune Receptors and T Cell Activation, Center for Pathophysiology, Infectiology, and Immunology, Institute of Immunology, Medical University of Vienna, Vienna, Austria.

Zlabinger 5, Peter Steinberger 1 * † and Johannes Hofer 4 * † Isenman 3, Markus Wahrmann 4, Judith Leitner 1, Gerhard J. Claire Battin 1, Annika De Sousa Linhares 1, Wolfgang Paster 1,2, David E.