TgPRELID, a Mitochondrial Protein Linked to Multidrug Resistance in the Parasite Toxoplasma gondii

New drugs to control infection with the protozoan parasite Toxoplasma gondii are needed as current treatments exert toxic side effects on patients. Approaches to develop novel compounds for drug development include screening of compound libraries and targeted inhibition of essential cellular pathways. We identified two distinct compounds that display inhibitory activity against the parasite's replicative stage: F3215-0002, which we previously identified during a compound library screen, and I-BET151, an inhibitor of bromodomains, the "reader" module of acetylated lysines. In independent studies, we sought to determine the targets of these two compounds using forward genetics, generating resistant mutants and identifying the determinants of resistance with comparative genome sequencing. Despite the dissimilarity of the two compounds, we recovered resistant mutants with nonsynonymous mutations in the same domain of the same gene, TGGT1_254250, which we found encodes a protein that localizes to the parasite mitochondrion (designated TgPRELID after the name of said domain). We found that mutants selected with one compound were cross resistant to the other compound, suggesting a common mechanism of resistance. To further support our hypothesis that TgPRELID mutations facilitate resistance to both I-BET151 and F3215-0002, CRISPR (clustered regularly interspaced short palindromic repeat)/CAS9-mediated mutation of TgPRELID directly led to increased F3215-0002 resistance. Finally, all resistance mutations clustered in the same subdomain of TgPRELID. These findings suggest that TgPRELID may encode a multidrug resistance factor or that I-BET151 and F3215-0002 have the same target(s) despite their distinct chemical structures. IMPORTANCE We report the discovery of TgPRELID, a previously uncharacterized mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. Drug resistance remains a major problem in the battle against parasitic infection, and understanding how TgPRELID mutations augment resistance to multiple, distinct compounds will reveal needed insights into the development of new therapies for toxoplasmosis and other related parasitic diseases

Multispecific Antibody Development Platform Based on Human Heavy Chain Antibodies

Heavy chain-only antibodies (HCAbs) do not associate with light chains and their VH regions are functional as single domains, forming the smallest active antibody fragment. These VH regions are ideal building blocks for a variety of antibody-based biologics because they tolerate fusion to other molecules and may also be attached in series to construct multispecific antibodies without the need for protein engineering to ensure proper heavy and light chain pairing. Production of human HCAbs has been impeded by the fact that natural human VH regions require light chain association and display poor biophysical characteristics when expressed in the absence of light chains. Here, we present an innovative platform for the rapid development of diverse sets of human HCAbs that have been selected in vivo. Our unique approach combines antibody repertoire analysis with immunization of transgenic rats, called UniRats, that produce chimeric HCAbs with fully human VH domains in response to an antigen challenge. UniRats express HCAbs from large transgenic loci representing the entire productive human heavy chain V(D)J repertoire, mount robust immune responses to a wide array of antigens, exhibit diverse V gene usage and generate large panels of stable, high affinity, antigen-specific molecules

Fabrication of La³⁺ Impregnated Chitosan/β-Cyclodextrin Biopolymeric Materials for Effective Utilization of Chromate and Fluoride Adsorption in Single Systems

Lanthanum impregnated chitosan/β-cyclodextrin (CS–La−βCD) composite was prepared using a facile <i>in situ</i> fabrication method and successfully evaluated for the adsorption of CrO₄^2– and F^–. The pristine and treated CS–La−βCD was characterized using Fourier transform infrared, scanning electron microscopy, energy dispersive X-ray spectroscopy with mapping, X-ray diffraction, and thermal gravametric analytical techniques. The optimum conditions for the uptake of CrO₄^2– and F^– were investigated as a function of shaking time, dosage, initial ion concentration, pH, and predatory ions in batch experiments. The equilibrium data were fitted using Langmuir, Freundlich, and Dubinin–Radushkevich models for the defluoridation and sorption of chromium onto CS–La−βCD. The adsorption kinetics of CrO₄^2– and F^– followed the pseudo-second order model. The maximum Langmuir adsorption capacity was found to be 91.58 mg/g and 8.14 mg/g for CrO₄^2– and F^–, respectively at 303 K. Sequential adsorption and desorption studies were carried out up to five cycles using 0.1 M of NaOH as an eluent to check the reusability of the adsorbents

Characterizations of a family of bivariate Pareto distributions

In the present paper, we study properties of a family of bivariate Pareto distributions. The well known dullness property of the univariate Pareto model is extended to the bivariate setup. Two measures of income inequality viz. income gap ratio and mean left proportional residual income are defined in the bivariate case. We also introduce bivariate generalized failure rate useful in reliability analysis. Characterizations, using the above concepts, for various members of the family of bivariate Pareto distributions are derived

A family of bivariate Pareto distributions

Pareto distributions have been extensively used in literature for modelling and analysis of income and lifetime data. In the present paper, we introduce a family of bivariate Pareto distributions using a generalized version of dullness property. Some important bivariate Pareto distributions are derived as special cases. Distributional properties of the family are studied. The dependency structure of the family is investigated. Finally, the family of distributions is applied to two real life data situatio

A bispecific antibody agonist of the IL-2 heterodimeric receptor preferentially promotes in vivo expansion of CD8 and NK cells

International audienceThe use of recombinant interleukin-2 (IL-2) as a therapeutic protein has been limited by significant toxicities despite its demonstrated ability to induce durable tumor-regression in cancer patients. The adverse events and limited efficacy of IL-2 treatment are due to the preferential binding of IL-2 to cells that express the high-affinity, trimeric receptor, IL-2Rαβγ such as endothelial cells and T-regulatory cells, respectively. Here, we describe a novel bispecific heavy-chain only antibody which binds to and activates signaling through the heterodimeric IL-2Rβγ receptor complex that is expressed on resting T-cells and NK cells. By avoiding binding to IL-2Rα, this molecule circumvents the preferential T-reg activation of native IL-2, while maintaining the robust stimulatory effects on T-cells and NK-cells in vitro. In vivo studies in both mice and cynomolgus monkeys confirm the molecule’s in vivo biological activity, extended pharmacodynamics due to the Fc portion of the molecule, and enhanced safety profile. Together, these results demonstrate that the bispecific antibody is a safe and effective IL-2R agonist that harnesses the benefits of the IL-2 signaling pathway as a potential anti-cancer therapy