Molecular characterization of a novel mitochondrial NOD-like receptor X1 in chicken that negatively regulates IFN-ß expression via STING.

NOD-like receptor X1 (NLRX1) is known for its unique mitochondrial localization and plays a negative role in innate immunity. The initial characterization and function of chicken NLRX1 remain unclear. Here, chicken mitochondrial-targeted NLRX1 (chNLRX1) protein was identified. It had relatively conserved domains, a unique N-terminal "X" mitochondrial-targeting domain (MT) and 2 highly conserved motifs at positions 510-520 and 412-421. Furthermore, chNLRX1 had a unique 53aa N-terminus-MT consistent with its localization to mitochondria. Additionally, chNLRX1 was observed to reduce the DNA sensing adaptor stimulator of interferon genes (STING)-induced IFN-ß by attenuating the STING-TANK-binding kinase 1 (TBK1) interaction, which is a requisite for the STING-TBK1-IFN-ß signaling pathway. These results suggested that chNLRX1 negatively regulated type-I interferon production via STING in host innate immunity.

Bioanalytical Assays for Pharmacokinetic and Biodistribution Study of Antibody-Drug Conjugates.

Antibody-drug conjugates (ADCs) are produced by the chemical linkage of cytotoxic agents and monoclonal antibodies. The complexity and heterogeneity of ADCs and the low concentration of cytotoxic agent released in vivo poses big challenges to their bioanalysis. Understanding the pharmacokinetic behavior, exposure-safety, and exposure-efficacy relationships of ADCs is needed for their successful development. Accurate analytical methods are required to evaluate intact ADCs, total antibody, released small molecule cytotoxins, and related metabolites. The selection of appropriate bioanalysis methods for comprehensive analysis of ADCs is mainly dependent on the properties of cytotoxic agents, the chemical linker, and the attachment sites. The quality of the information about the whole pharmacokinetic profile of ADCs has been improved due to the development and improvement of analytical strategies for detection of ADCs, such as ligand-binding assays and mass spectrometry-related techniques. In this article, we will focus on the bioanalytical assays that have been used in the pharmacokinetic study of ADCs and discuss their advantages, current limitations, and potential challenges. SIGNIFICANCE STATEMENT: This article describes bioanalysis methods which have been used in pharmacokinetic study of ADCs and discusses the advantages, disadvantages and potential challenges of these assays. This review is useful and helpful and will provide insights and reference for bioanalysis and development of ADCs.

Chicken anemia virus VP1 negatively regulates type I interferon via targeting interferon regulatory factor 7 of the DNA-sensing pathway.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway plays a vital role in sensing viral DNA in the cytosol, stimulating type I interferon (IFN) production and triggering the innate immune response against DNA virus infection. However, viruses have evolved effective inhibitors to impede this sensing pathway. Chicken anemia virus (CAV), a nonenveloped ssDNA virus, is a ubiquitous pathogen causing great economic losses to the poultry industry globally. CAV infection is reported to downregulate type I IFN induction. However, whether the cGAS-STING signal axis is used by CAV to regulate type I IFN remains unclear. Our results demonstrate that CAV infection significantly elevates the expression of cGAS and STING at the mRNA level, whereas IFN-ß levels are reduced. Furthermore, IFN-ß activation was completely blocked by the structural protein VP1 of CAV in interferon stimulatory DNA (ISD) or STING-stimulated cells. VP1 was further confirmed as an inhibitor by interacting with interferon regulatory factor 7 (IRF7) by binding its C-terminal 143-492 aa region. IRF7 dimerization induced by TANK binding kinase 1 (TBK1) could be inhibited by VP1 in a dose-dependent manner. Together, our study demonstrates that CAV VP1 is an effective inhibitor that interacts with IRF7 and antagonizes cGAS-STING pathway-mediated IFN-ß activation. These findings reveal a new mechanism of immune evasion by CAV.

Nitrogen fertilization and CO2 concentration synergistically affect the growth and protein content of Agropyron mongolicum.

Background: The nitrogen (N) and protein concentrations in plant tissues exposed to elevated CO2 (eCO2) generally decline , such declines in forage grass composition are expected to have negative implications for the nutritional and economic value of grass. Plants require N for the production of a photosynthetically active canopy and storage proteins in the tissues, whose functionality will strongly influence productivity and quality. The objective of this study was to investigate whether eCO2 plus N-fertilization increases growth and N nutrition of Agropyron mongolicum, and the dependence of this improvement on the coordination between root and leaf development. Methods: We analyzed A. mongolicum from field-grown within the open-top chambers (OTCs) facility under two atmospheric CO2 (ambient, 400 ± 20 µmol mol-1, aCO2, and elevated, 800 ± 20 µmol mol-1, eCO2) and three N-fertigation treatments (control, low N-fertigation , and high N-fertigation) for two months. Results: Elevated CO2 plus N-fertigation strongly increased shoot and root biomass, and the nitrogen and protein concentrations of A. mongolicum compared to those plants at aCO2 levels. Increased N content in leaves and reduced specific leaf area (SLA) at a high N supply could alleviate photosynthetic acclimation to eCO2 and drive the production of greater shoot biomass with the potential for higher photosynthesis, productivity, and nutritional quality. The increased root length (RL), the ratio of total aboveground N taken up per RL (TN/RL), stomatal conductance (Gs), and transpiration rate (Tr) contribute to the transpiration-driven mass flow of N, consequently increasing N uptake by roots. In addition, a smaller percentage of N remained as unassimilated nitrate ( NO 3 - ) under eCO2, indicating that assimilation of NO 3 - into proteins was not inhibited by eCO2. These findings imply that grass productivity and quality will enhance under anticipated elevated CO2 concentration when effective management measures of N-fertilization are employed.

Inhibition of the DNA-Sensing pathway by pseudorabies virus UL24 protein via degradation of interferon regulatory factor 7.

The cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway plays an important role in the innate immune response by the production of type I interferon (IFN) against DNA virus infection. However, viruses have evolved a variety of strategies to antagonize the host antiviral response to facilitate infection and replication. Pseudorabies virus (PRV), a DNA virus that causes great economic losses to the swine industry, encodes approximate 70 proteins, including some that are involved in evasion of host immunity. However, the mechanism employed by PRV to regulate type I IFN remains unclear. The results of the present study showed that the transcription levels of type I IFN were significantly upregulated by a UL24-deleted PRV strain. Furthermore, IFN-ß activation induced by poly(dA:dT) or stimulated by cGAS-STING was inhibited by UL24 overexpression in PK15 cells. Co-immunoprecipitation analysis demonstrated that UL24 interacts with and can degrade interferon regulatory factor 7 (IRF7) through the proteasome pathway in a dose-dependent manner. Together, these results showed that PRV UL24 interacted with IRF7 via the proteasome pathway and antagonized cGAS-STING-mediated activation of IFN-ß.