“
“The glycan shield of the human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein serves as a barrier to antibody-mediated neutralization and plays a critical role in transmission and infection. One of the few broadly neutralizing HIV-1 antibodies, 2G12, binds to a carbohydrate epitope consisting of an array of high-mannose glycans exposed on the surface of the gp120 subunit of the Env protein. To produce proteins with exclusively high-mannose carbohydrates, we generated a mutant strain of Saccharomyces cerevisiae by deleting three genes in the N-glycosylation pathway, Och1, Mnn1, and Mnn4.

Glycan profiling revealed that N-glycans produced by this mutant were almost exclusively Man(8)GlcNAc(2), and four endogenous glycoproteins learn more that were efficiently recognized by the 2G12 antibody were identified. These yeast proteins, like HIV-1 gp120, contain a large

number and high density of N-linked glycans, with glycosidase digestion abrogating 2G12 cross-reactivity. Immunization of rabbits with whole Delta och1 Delta mnn1 Delta mnn4 yeast cells produced sera that recognized a broad range of HIV-1 and simian immunodeficiency virus (SIV) Env glycoproteins, despite no HIV/SIV-related proteins being used in the immunization procedure. Analyses of one of these sera on a glycan array showed strong binding to glycans with terminal Man alpha 1,2Man residues, and binding to gp120 was abrogated by glycosidase removal BAY 1895344 of high-mannose glycans and terminal Man alpha 1,2Man residues, similar to 2G12. Since S. cerevisiae is genetically pliable and can be grown easily and inexpensively, it will be possible to produce new immunogens that recapitulate the 2G12 epitope and may make the glycan shield of HIV Env a practical target for vaccine development.”
“The thalamocortical selleck chemicals (TC) projection in the mammalian brain involves fundamental

aspects in branch formation during development. TC axons are known to form branches not only in a genetically defined but also in an activity-dependent fashion. Recent evidence indicates that TC axon branching is generated by positive and negative regulators that are expressed with laminar specificity in the developing cortex. Moreover, in vitro studies using organotypic cocultures demonstrate that neural activity, including firing and synaptic activity, controls lamina-specific TC axon branching by altering its remodeling process with addition and elimination. Taken together, activity-dependent mechanisms can contribute to branch formation, affecting expression of branch-promoting and inhibiting factors and/or their receptor molecules.