Expression of the readthrough transcript CiDRE in alveolar macrophages boosts SARS-CoV-2 susceptibility and promotes COVID-19 severity.

Lung infection during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) via the angiotensin-I-converting enzyme 2 (ACE2) receptor induces a cytokine storm. However, the precise mechanisms involved in severe COVID-19 pneumonia are unknown. Here, we showed that interleukin-10 (IL-10) induced the expression of ACE2 in normal alveolar macrophages, causing them to become vectors for SARS-CoV-2. The inhibition of this system in hamster models attenuated SARS-CoV-2 pathogenicity. Genome-wide association and quantitative trait locus analyses identified a IFNAR2-IL10RB readthrough transcript, COVID-19 infectivity-enhancing dual receptor (CiDRE), which was highly expressed in patients harboring COVID-19 risk variants at the IFNAR2 locus. We showed that CiDRE exerted synergistic effects via the IL-10-ACE2 axis in alveolar macrophages and functioned as a decoy receptor for type I interferons. Collectively, our data show that high IL-10 and CiDRE expression are potential risk factors for severe COVID-19. Thus, IL-10R and CiDRE inhibitors might be useful COVID-19 therapies.

The dCache Domain of the Chemoreceptor Tlp1 in Campylobacter jejuni Binds and Triggers Chemotaxis toward Formate.

Chemotaxis is an important virulence factor in some enteric pathogens, and it is involved in the pathogenesis and colonization of the host. However, there is limited knowledge regarding the environmental signals that promote chemotactic behavior and the sensing of these signals by chemoreceptors. To date, there is no information on the ligand molecule that directly binds to and is sensed by Campylobacter jejuni Tlp1, which is a chemoreceptor with a dCache-type ligand-binding domain (LBD). dCache (double Calcium channels and chemotaxis receptor) is the largest group of sensory domains in bacteria, but the dCache-type chemoreceptor that directly binds to formate has not yet been discovered. In this study, formate was identified as a direct-binding ligand of C. jejuni Tlp1 with high sensing specificity. We used the strategy of constructing a functional hybrid receptor of C. jejuni Tlp1 and the Escherichia coli chemoreceptor Tar to screen for the potential ligand of Tlp1, with the binding of formate to Tlp1-LBD being verified using isothermal titration calorimetry. Molecular docking and experimental analyses indicated that formate binds to the membrane-proximal pocket of the dCache subdomain. Chemotaxis assays demonstrated that formate elicits robust attractant responses of the C. jejuni strain NCTC 11168, specifically via Tlp1. The chemoattraction effect of formate via Tlp1 promoted the growth of C. jejuni, especially when competing with Tlp1- or CheY-knockout strains. Our study reveals the molecular mechanisms by which C. jejuni mediates chemotaxis toward formate, and, to our knowledge, is the first report on the high-specificity binding of the dCache-type chemoreceptor to formate as well as the physiological role of chemotaxis toward formate. IMPORTANCE Chemotaxis is important for Campylobacter jejuni to colonize favorable niches in the gastrointestinal tract of its host. However, there is still a lack of knowledge about the ligand molecules for C. jejuni chemoreceptors. The dCache-type chemoreceptor, namely, Tlp1, is the most conserved chemoreceptor in C. jejuni strains; however, the direct-binding ligand(s) triggering chemotaxis has not yet been discovered. In the present study, we found that the ligand that binds directly to Tlp1-LBD with high specificity is formate. C. jejuni exhibits robust chemoattraction toward formate, primarily via Tlp1. Tlp1 is the first reported dCache-type chemoreceptor that specifically binds formate and triggers strong chemotaxis. We further demonstrated that the formate-mediated promotion of C. jejuni growth is correlated with Tlp1-mediated chemotaxis toward formate. Our work provides important insights into the mechanism and physiological function of chemotaxis toward formate and will facilitate further investigations into the involvement of microbial chemotaxis in pathogen-host interactions.

Cell cycle control by the insulin-like growth factor signal: at the crossroad between cell growth and mitotic regulation.

In proliferating cells and tissues a number of checkpoints (G1/S and G2/M) preceding cell division (M-phase) require the signal provided by growth factors present in serum. IGFs (I and II) have been demonstrated to constitute key intrinsic components of the peptidic active fraction of mammalian serum. In vivo genetic ablation studies have shown that the cellular signal triggered by the IGFs through their cellular receptors represents a non-replaceable requirement for cell growth and cell cycle progression. Retroactive and current evaluation of published literature sheds light on the intracellular circuitry activated by these factors providing us with a better picture of the pleiotropic mechanistic actions by which IGFs regulate both cell size and mitogenesis under developmental growth as well as in malignant proliferation. The present work aims to summarize the cumulative knowledge learned from the IGF ligands/receptors and their intracellular signaling transducers towards control of cell size and cell-cycle with particular focus to their actionable circuits in human cancer. Furthermore, we bring novel perspectives on key functional discriminants of the IGF growth-mitogenic pathway allowing re-evaluation on some of its signal components based upon established evidences.

Fabrication of a novel electrochemical biosensor based on a molecular imprinted polymer-aptamer hybrid receptor for lysozyme determination.

In this work, a novel, sensitive, and rapid electrochemical biosensor was employed to detect lysozyme (Lys) using a double receptor of molecular imprinted polymer (MIP)-aptamer. First, a glassy carbon electrode (GCE) was modified with a nanocomposite consisting of multi-wall carbon nanotubes (MWCNTs), nitrogen-doped carbon quantum dots (N-CQDs), and chitosan. Subsequently, aptamer (Apt)-Lys complex was immobilized on MWCNTs-N-CQDs-chitosan/GCE via binding between carboxyl groups present in the nanocomposite and the terminal amine groups of the aptamer. Following that, methylene blue monomer was electrochemically polymerized around the Apt-Lys complex on the MWCNTs-N-CQDs-chitosan/GCE surface. Finally, after the template removal, the remaining cavities along with the aptamers created a new hybrid receptor of MIP-aptamer. The MWCNTs-N-CQDs-chitosan nanocomposite could provide large amounts of carboxyl groups for binding to amino-functionalized aptamers, considerable electrical conductivity, and a high surface-to-volume ratio. These beneficial features facilitated the Apt-Lys complex immobilization and gave improved electrochemical signal. The obtained MIP-aptamer hybrid receptor allowed lysozyme determination even at concentrations as low as 4.26 fM within the functional range of 1 fM to 100 nM.

How insulin-like growth factor I binds to a hybrid insulin receptor type 1 insulin-like growth factor receptor.

Monomers of the insulin receptor and type 1 insulin-like growth factor receptor (IGF-1R) can combine stochastically to form heterodimeric hybrid receptors. These hybrid receptors display ligand binding and signaling properties that differ from those of the homodimeric receptors. Here, we describe the cryoelectron microscopy structure of such a hybrid receptor in complex with insulin-like growth factor I (IGF-I). The structure (ca. 3.7 Å resolution) displays a single IGF-I ligand, bound in a similar fashion to that seen for IGFs in complex with IGF-1R. The IGF-I ligand engages the first leucine-rich-repeat domain and cysteine-rich region of the IGF-1R monomer (rather than those of the insulin receptor monomer), consistent with the determinants for IGF binding residing in the IGF-1R cysteine-rich region. The structure broadens our understanding of this receptor family and assists in delineating the key structural motifs involved in binding their respective ligands.

[Source Apportionment of Air Pollutants for a Typical Pollution Event in Zhaoqing].

Based on observational data for pollutants and meteorology, this study analyzed the pollution episode that occurred during Dec 17th to 23th in 2018 in Zhaoqing, Guangdong Province, China. Using the source apportionment model CMAQ-ISAM and the hybrid receptor model, the regional contributions to air pollution were examined. The results showed that low-pressure conditions had an adverse effect on the diffusion of pollutants during this pollution episode in Zhaoqing. Prior to the pollution episode, pollutants were mainly derived from Zhaoqing and Qingyuan, accounting for 19.2% and 10.7% of pollutants, respectively. As well as pollutants from Guangdong Province, long-distance transport of pollutants from Jiangxi, Hunan, Hubei, and Shaanxi accounted for approximately 64.5% of the total during the non-pollution period. During the polluted episode, major cities in Pearl River Delta and the eastern part of Guangdong Province contributed more pollutants as a surface high-pressure field moved southward. Zhaoqing, Foshan, Dongguan, Guangzhou, and Huizhou contributed 25.5%, 14.8%, 9.8%, 9.5%, and 5.3% of the pollutants, respectively. Cities in the eastern part of Guangdong Province including Heyuan, Meizhou, Shanwei, Jieyang, Shantou, and Chaozhou contributed 13.7% of the total pollutants. In addition, pollutants from Fujian, Jiangxi, and the Yangtze River Delta accounted for approximately 32.9%. Furthermore, pollutants transported under marine influences were one of the main causes of this pollution episode in Zhaoqing.

Approaches for identifying PM2.5 source types and source areas at a remote background site of South China in spring.

The receptor model is an effectively and widely used tool for analyzing the source of PM2.5, and its development and improvement have always been focused and challenged. In this study, approaches of source analysis is applied and compared. The PM2.5 samples were collected in spring of 2015 at a remote background site of Weizhou, South China and were analyzed for water-soluble ions, trace metals, and sugars. The 28 measurement species were introduced into the positive matrix factorization (PMF) and a non-negative matrix factorization (NMF) model for inter-comparison of PM2.5 prediction. Results showed that the NMF model is a more robust tool to identify source types and source apportionment in the case of a small sample size (n = 31). In NMF, four source variants were obtained as dust (15.6%), biomass combustion (11.8%), secondary formation (17.6%), and coal combustion (54.9%), corresponding to four main source areas. These were Southeast Asia, South China Sea, Taiwan Strait, as well as Pearl River Delta, respectively. The areas were distinguished based on hybrid receptor models, potential source contribution function (PSCF) and concentration weighted trajectory (CWT), by introducing the daily loadings of each source factor from NMF method. These model results were highly consistent with categorized chemical characteristics of PM2.5, suggesting that NMF linking with hybrid receptor models provides valuable implications for exploring source types and source areas of PM2.5. Meanwhile, biomass combustion and coal combustion comparably contributed to the high PM2.5 concentrations indicating control strategy in South China in spring.

Cross-species reactive monoclonal antibodies against the extracellular domains of the insulin receptor and IGF1 receptor.

Translation across species of immunoassay results is often challenging due to the lack of cross-species reactivity of antibodies. In order to investigate the biology of insulin and IGF1 receptors, we generated new versatile monoclonal assay antibodies using the extracellular domain of the insulin/IGF1 hybrid receptor as the bait protein in the Adimab yeast antibody discovery platform and as the antigen in a rabbit monoclonal antibody platform. The resulting antibody clones were screened for receptor specificity as well as cross-species reactivity to both tissue and cell line derived samples. Using these strategies, we were able to identify highly specific insulin receptor monoclonal antibodies that lack cross-reactivity to the IGF1 receptor using the Adimab platform and a highly specific IGF1 receptor monoclonal antibody that lacks cross-reactivity to the insulin receptor using the rabbit antibody platform. Unlike earlier monoclonal antibodies reported in the literature, these antibodies show cross-species reactivity to the extracellular domains of mouse, rat, pig, and human receptors, indicating that they bind conserved epitopes. Furthermore, the antibodies work well in several different assay formats, including ELISA, flow cytometry, and immunoprecipitation, and therefore provide new tools to study insulin and IGF1 receptor biology with translation across several species and experimental model systems.

Building the Case for Insulin-Like Growth Factor Receptor-I Involvement in Thyroid-Associated Ophthalmopathy.

The pathogenesis of orbital Graves' disease (GD), a process known as thyroid-associated ophthalmopathy (TAO), remains incompletely understood. The thyrotropin receptor (TSHR) represents the central autoantigen involved in GD and has been proposed as the thyroid antigen shared with the orbit that could explain the infiltration of immune cells into tissues surrounding the eye. Another cell surface protein, insulin-like growth factor-I receptor (IGF-IR), has recently been proposed as a second antigen that participates in TAO by virtue of its interactions with anti-IGF-IR antibodies generated in GD, its apparent physical and functional complex formation with TSHR, and its necessary involvement in TSHR post-receptor signaling. The proposal that IGF-IR is involved in TAO has provoked substantial debate. Furthermore, several studies from different laboratory groups, each using different experimental models, have yielded conflicting results. In this article, we attempt to summarize the biological characteristics of IGF-IR and TSHR. We also review the evidence supporting and refuting the postulate that IGF-IR is a self-antigen in GD and that it plays a potentially important role in TAO. The putative involvement of IGF-IR in disease pathogenesis carries substantial clinical implications. Specifically, blocking this receptor with monoclonal antibodies can dramatically attenuate the induction by TSH and pathogenic antibodies generated in GD of proinflammatory genes in cultured orbital fibroblasts and fibrocytes. These cell types appear critical to the development of TAO. These observations have led to the conduct of a now-completed multicenter therapeutic trial of a fully human monoclonal anti-IGF-IR blocking antibody in moderate to severe, active TAO.

Surface-expressed insulin receptors as well as IGF-I receptors both contribute to the mitogenic effects of human insulin and its analogues.

There is a medical need for new insulin analogues. Yet, molecular alterations to the insulin molecule can theoretically result in analogues with carcinogenic effects. Preclinical carcinogenicity risk assessment for insulin analogues rests to a large extent on mitogenicity assays in cell lines. We therefore optimized mitogenicity assay conditions for a panel of five cell lines. All cell lines expressed insulin receptors (IR), IGF-I receptors (IGF-IR) and hybrid receptors, and in all cell lines, insulin as well as the comparator compounds X10 and IGF-I caused phosphorylation of the IR as well as IGF-IR. Insulin exhibited mitogenicity EC(50) values in the single-digit nanomolar to picomolar range. We observed correlations across cell types between (i) mitogenic potency of insulin and IGF-IR/IR ratio, (ii) Akt phosphorylation and mitogenic potency and (iii) Akt phosphorylation and IR phosphorylation. Using siRNA-mediated knockdown of IR and IGF-IR, we observed that in HCT 116 cells the IR appeared dominant in driving the mitogenic response to insulin, whereas in MCF7 cells the IGF-IR appeared dominant in driving the mitogenic response to insulin. Together, our results show that the IR as well as IGF-IR may contribute to the mitogenic potency of insulin. While insulin was a more potent mitogen than IGF-I in cells expressing more IR than IGF-IR, the hyper-mitogenic insulin analogue X10 was a more potent mitogen than insulin across all cell types, supporting that the hyper-mitogenic effect of X10 involves the IR as well as the IGF-IR. These results are relevant for preclinical safety assessment of developmental insulin analogues.