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Dalton, V.

Publications and source records attributed to Dalton, V..

2 recordsLinked to original sources

Whole-body PET imaging of SIV using anti-Env probes fails to reveal regions of specific uptake in rhesus macaques

Following the initial reports demonstrating the feasibility of immunoPET imaging of SIV using anti-Env monoclonal antibodies in non-human primates, replication efforts of the imaging system in HIV-infected individuals have yielded conflicting results. Herein, we used 89Zr-7D3 and 89Zr- ITS103.01LS-F(ab)2, two anti-gp120 antibodies for immunoPET imaging of SIV in n=20 rhesus macaques. Despite their demonstrated nanomolar affinity and strong binding specificity to SIV gp120 cell lines, we observed no discernible differences in their binding in primary cells, tissue sections of secondary lymphoid organs, in-vivo probe uptake between SIVmac-infected and uninfected macaques, or ex-vivo validation necropsies. While the probes remained stable in-vivo, only 89Zr-ITS103.01LS-F(ab)2 in chronic plasma retained its binding specificity to SIV gp120, with 89Zr-7D3 experiencing a >97% reduction in binding to gp120 due to competition from endogenous antibodies at the 7D3 binding site. The overall absence of specific uptake suggests inadequate binding potential (ligand affinity x target molarity) for these probes to effectively image SIV or HIV in-vivo, warranting further investigation into the lack of reproducibility observed with earlier non-human primate SIV imaging and conflicting human studies.

immunology↗

Full-length αIIbβ3 CryoEM structure reveals intact integrin initiate-activation intrinsic architecture

Integrin IIb{beta}3 is the key receptor regulating platelet retraction and accumulation, thus pivotal for hemostasis, and arterial thrombosis as well as a proven drug-target for antithrombotic therapies. Here we resolve the cryoEM structures of the intact full-length IIb{beta}3, which covers three distinct states along the activation pathway. Here, we resolve intact IIb{beta}3 structure at 3[A] resolution, revealing the overall topology of the heterodimer with the transmembrane (TM) helices and the head region ligand-binding domain tucked in a specific angle proximity to the TM region. In response to the addition of a Mn2+ agonist, we resolved two coexisting states, "intermediate" and "pre-active". Our structures show conformational changes of the intact IIb{beta}3 activating trajectory, as well as a unique twisting of the lower integrin legs representing an intermediate state (TM region at a twisting conformation) and a coexisting pre-active state (bent and opening in leg), which is required for inducing the transitioning platelets to accumulate. Our structure provides for the first time direct structural evidence for the lower legs involvement in full-length integrin activation mechanisms. Additionally, our structure offers a new strategy to target the IIb{beta}3 lower leg allosterically instead of modulating the affinity of the IIb{beta}3 head region.

biochemistry↗