Delineating the cellular signaling networks modulated by these vGPCRs will be crucial for treatment of virus-associated pathologies. (C) 2010 Elsevier Ireland Ltd. All rights reserved.”
“Background: Ten to 30% of HIV-1 infected subjects develop broadly neutralizing antibodies (bNAbs) during chronic infection. We hypothesized that immunizing rabbits with viral envelope glycoproteins (Envs) from ABT-263 manufacturer these patients may induce bNAbs, when formulated as a trimeric protein and in the presence of an adjuvant.\n\nMethods:

Based on in vitro neutralizing activity in serum, patients with bNAbs were selected for cloning of their HIV-1 Env. Seven stable soluble trimeric gp140 proteins were generated from sequences derived from four adults and two children infected with either clade A or B HIV-1. From one of the clade A Envs both the monomeric and trimeric Env were produced for comparison. Rabbits were immunized with soluble gp120 or trimeric gp140 proteins Volasertib Cell Cycle inhibitor in combination with the adjuvant dimethyl dioctadecyl ammonium/trehalose dibehenate (CAF01). Env binding in rabbit immune serum was determined using ELISAs based on gp120-IIIB protein. Neutralizing activity of IgG purified from rabbit immune sera was measured with the pseudovirus-TZMbl assay and a PBMC-based neutralization assay for selected experiments.\n\nResults: It was initially established that gp140 trimers induce better antibody

responses over gp120 monomers and that the adjuvant CAF01 was necessary for such strong responses. Gp140 trimers, based on HIV-1 variants from patients with bNAbs, were able to elicit both gp120(IIIB) specific IgG and NAbs to Tier 1 viruses of different subtypes. Potency of NAbs closely correlated with titers, and an gp120-binding IgG titer above a threshold of 100,000 was predictive of neutralization selleck kinase inhibitor capability. Finally, peptide inhibition experiments showed that a large fraction of the neutralizing IgG was directed against the gp120 V3 region.\n\nConclusions: Our results indicate that the strategy of reverse immunology based on selected Env sequences

is promising when immunogens are delivered as stabilized trimers in CAF01 adjuvant and that the rabbit is a valuable model for HIV vaccine studies.”
“Background: Domain antibodies (dAbs; similar to 10-15 kDa) are made up of the variable heavy chain or the variable light chain of the antibody structure, and retain binding capability, dAbs have proved difficult to detect in plasma using immunoassay without specific antibodies raised against the dAb. Results: A sensitive and selective UPLC-MS/MS method for the absolute quantification of a dAb in monkey plasma was developed (range: 1 to 500 ng/ml) without the need for a specific capture antibody. This method was used to analyze pharmacokinetic studies early on in drug development. Furthermore, an immunoassay was developed and the pharmacokinetic samples were reanalyzed.