Ісак Мазепа: перші кроки у великій політиці

Досліджується становлення світоглядних позицій та формування політичних поглядів І. Мазепи.Исследуется становление мировоззренческих позиций и формирование политических взглядов И. Мазепы.Formation world outlook positions and formation of political views of I. Mazepa is investigated

A bispecific T cell engager recruits both type 1 NKT and Vy9V52-T cells for the treatment of CD1d-expressing hematological malignancies

Bispecific T cell engagers (bsTCEs) hold great promise for cancer treatment but face challenges due to the induction of cytokine release syndrome (CRS), on-target off-tumor toxicity, and the engagement of immuno-suppressive regulatory T cells that limit efficacy. The development of Vy9V52-T cell engagers may overcome these challenges by combining high therapeutic efficacy with limited toxicity. By linking a CD1d-specific single-domain antibody (VHH) to a V52-TCR-specific VHH, we create a bsTCE with trispecific properties, which engages not only Vy9V52-T cells but also type 1 NKT cells to CD1d+ tumors and triggers robust proin-flammatory cytokine production, effector cell expansion, and target cell lysis in vitro. We show that CD1d is expressed by the majority of patient MM, (myelo)monocytic AML, and CLL cells and that the bsTCE triggers type 1 NKT and Vy9V52-T cell-mediated antitumor activity against these patient tumor cells and improves survival in in vivo AML, MM, and T-ALL mouse models. Evaluation of a surrogate CD1d-y5 bsTCE in NHPs shows Vy9V52-T cell engagement and excellent tolerability. Based on these results, CD1d-V52 bsTCE (LAVA-051) is now evaluated in a phase 1/2a study in patients with therapy refractory CLL, MM, or AML.Transplantation and autoimmunit

Construction and characterization of a bispecific diabody for retargeting T cells to human carcinomas

We describe the construction of a recombinant bispecific antibody fragment in the diabody format with specificity for both the well-established human pancarcinoma associated target antigen EGP2 (epithelial glycoprotein 2, also known as the CO 17-1A antigen or KSA) and the CD3 epsilon chain of human TCR/CD3 complex. The murine anti-ECP2 (MOC31) single chain variable fragment (scFv) and the humanized anti-CD3 (Ucht1v9) scFv were cast into a diabody format (designated Dia5v9) using a short 5 amino acid Gly-Ser linker between immunoglobulin heavy-chain and light-chain variable domains. Purification of the poly-histidine tagged Dia5v9 was achieved from extracts of protease deficient Escerichia coli by IMAC chromatography. The Dia5v9 diabody showed strong binding to both EGP2 and CD3 in transfected cells. The in vitro efficacy of Dia5v9 in mediating tumor cell lysis by interleukin-2 activated human T cells appeared to be similar to that of the hybrid-hybridoma-derived BsF(ab')(2) Bis1 (anti-EGP2/anti-CD3) in a standard 4-hr Cr-51-release assay. This small and partially humanized recombinant bispecific antibody fragment may be valuable for T-cell-based immuno-therapeutical treatment protocols, retargeting activated peripheral blood T lymphocytes to lyse various human carcinomas in vivo. (C) 1998 Wiley-Liss, Inc.</p

Endocytosis of EGFR requires its kinase activity and N-terminal transmembrane dimerization motif

EGFR signaling is attenuated by endocytosis and degradation of receptor-ligand complexes in lysosomes. Endocytosis of EGFR is known to be regulated by multiple post-translational modifications. The observation that prevention of these modifications does not block endocytosis completely, suggests the involvement of other mechanism(s). Recently, receptor clustering has been suggested to induce internalization of multiple types of membrane receptors. However, the mechanism of clustering-induced internalization remains unknown. We have used biparatopic antibody fragments from llama (VHHs) to induce EGFR clustering without stimulating tyrosine kinase activity. Using this approach, we have found an essential role for the N-terminal GG4-like dimerization motif in the transmembrane domain (TMD) for clustering-induced internalization. Moreover, conventional EGF-induced receptor internalization depends exclusively on this TMD dimerization and kinase activity. Mutations in this dimerization motif eventually lead to reduced EGFR degradation and sustained signaling. We propose a novel role for the TMD dimerization motif in the negative-feedback control of EGFR. The widely conserved nature of GG4-like dimerization motifs in transmembrane proteins suggests a general role for these motifs in clustering-induced internalization

Rapid Visualization of Human Tumor Xenografts through Optical Imaging with a Near-infrared Fluorescent Anti-Epidermal Growth Factor Receptor Nanobody

Given that overexpression of the epidermal growth factor receptor (EGFR) is found in many types of human epithelial cancers, noninvasive molecular imaging of this receptor is of great interest. A number of studies have employed monoclonal antibodies as probes; however, their characteristic long half-life in the bloodstream has encouraged the development of smaller probes. In this study, an anti-EGFR nanobody-based probe was developed and tested in comparison with cetuximab for application in optical molecular imaging. To this aim, the anti-EGFR nanobody 7D12 and cetuximab were conjugated to the near-infrared fluorophore IRDye800CW. 7D12-IR allowed the visualization of tumors as early as 30 minutes postinjection, whereas with cetuximab-IR, no signal above background was observed at the tumor site. Quantification of the IR-conjugated proteins in the tumors revealed ≈ 17% of injected dose per gram 2 hours after injection of 7D12-IR, which was significantly higher than the tumor uptake obtained 24 hours after injection of cetuximab-IR. This difference is associated with the superior penetration and distribution of 7D12-IR within the tumor. These results demonstrate that this anti-EGFR nanobody conjugated to the NIR fluorophore has excellent properties for rapid preclinical optical imaging, which holds promise for its future use as a complementary diagnostic tool in humans

Subalpine forest development following a blowdown in the Mount Zirkel Wilderness, Colorado

We documented the occurrence of a 1934 blowdown in a subalpine forest in northwestern Colorado, USA. Prior to the blowdown, the stand was dominated by old-growth Picea engelmannii - Abies lasiocarpa forests. Although blowdowns are believed to trigger outbreaks of Dendroctonus rufipennis (spruce beetle), we found no detectable increase in beetle caused mortality. Forest recovery was by both release of the previously suppressed regeneration and by new seedling establishment. Both recovery pathways were dominated by Abies. The blowdown thus caused a shift in species dominance from Picea to Abies; 65 yr after the blowdown, the fallen logs and tip up mounds continue to provide favourable habitat for seedling establishment of both species. The present study shows that the legacy of blowdowns can influence forest dynamics for decades following the disturbance event