Comparative Description of the Expression Profile of Interferon-Stimulated Genes in Multiple Cell Lineages Targeted by HIV-1 Infection

Immediately after viral infections, innate immune sensors recognize viruses and lead to the production of type I interferon (IFN-I). IFN-I upregulates various genes, referred to as IFN-stimulated genes (ISGs), and some ISGs inhibit viral replication. HIV-1, the causative agent of AIDS, mainly infects CD4+ T cells and macrophages and triggers the IFN-I-mediated signaling cascade. Certain ISGs are subsequently upregulated by IFN-I stimulus and potently suppress HIV-1 replication. HIV-1 cell biology has shed light on the molecular understanding of the IFN-I production triggered by HIV-1 infection and the antiviral roles of ISGs. However, the differences in the gene expression patterns following IFN-I stimulus among HIV-1 target cell types are poorly understood. In this study, we hypothesize that the expression profiles of ISGs are different among HIV-1 target cells and address this question by utilizing public transcriptome datasets and bioinformatic techniques. We focus on three cell types intrinsically targeted by HIV-1, including CD4+ T cells, monocytes, and macrophages, and comprehensively compare the expression patterns of ISGs among these cell types. Furthermore, we use the datasets of the differentially expressed genes by HIV-1 infection and the evolutionarily conserved ISGs in mammals and perform comparative transcriptome analyses. We defined 104 ‘common ISGs’ that were upregulated by IFN-I stimulus in CD4+ T cells, monocytes, and macrophages. The ISG expression patterns were different among these three cell types, and intriguingly, both the numbers and the magnitudes of upregulated ISGs by IFN-I stimulus were greatest in macrophages. We also found that the upregulated genes by HIV-1 infection included most ‘common ISGs.’ Moreover, we determined that the ‘common ISGs,’ particularly those with antiviral activity, were evolutionarily conserved in mammals. To our knowledge, this study is the first investigation to comprehensively describe (i) the different expression patterns of ISGs among HIV-1 target cells, (ii) the overlap in the genes modulated by IFN-I stimulus and HIV-1 infection and (iii) the evolutionary conservation in mammals of the antiviral ISGs that are expressed in HIV-1 target cells. Our results will be useful for deeply understanding the relationship of the effect of IFN-I and the modulated gene expression by HIV-1 infection

陸上競技の種目特性が大学女子選手の膝関節固有感覚に及ぼす影響

The objective of this study was to clarify the characteristics of knee joint proprioception in healthy female athletes. The subjects were 20 female college athletes with no serious medical history in the knee joints（jumping event group, n=10; non-jumping event group, n=10）. There was no significant difference between the groups in regard to physical characteristics, athletic history, and isokinetic leg muscle strength. The objective and methods of the study were fully explained to all subjects, and consent to participate was obtained. Knee joint position sense was measured with low （knee flexion 15-30°） and high （knee flexion 60-75°） flexion angle ranges. Knee joint kinesthesia was measured by bending 1 knee without prior warning from a stationary state with the knee joint in a slightly bent position （20-30°）. ANOVA was performed for statistical analysis and a multiple comparison test was conducted. For both methods, statistical significance was defined as 5％（p<0.05）. No significant difference was observed for position sense between the groups for the low and high flexion ranges. In kinesthesia, the dominant leg of the athletes in the jumping event group had a significantly smaller value than both the dominant and non-dominant legs of the athletes in the non-jumping event group （p<0.05, p<0.01 respectively）. Also, a significant difference（p<0.01） was observed between the dominant leg of the athletes in the jumping event group and that of the athletes in the non-jumping event group. There were no significant differences between the jumping and non-jumping event groups in regard to physical characteristics, athletic history, and isokinetic leg muscle strength. We concluded that the characteristics of the events were related to the fact that the jumping event group had better kinesthesia scores than the non-jumping event group

Comparative study on the effect of human BST-2/Tetherin on HIV-1 release in cells of various species

In this study, we first demonstrate that endogenous hBST-2 is predominantly expressed on the plasma membrane of a human T cell line, MT-4 cells, and that Vpu-deficient HIV-1 was less efficiently released than wild-type HIV-1 from MT-4 cells. In addition, surface hBST-2 was rapidly down-regulated in wild-type but not Vpu-deficient HIV-1-infected cells. This is a direct insight showing that provirus-encoded Vpu has the potential to down-regulate endogenous hBST-2 from the surface of HIV-1-infected T cells. Corresponding to previous reports, the aforementioned findings suggested that hBST-2 has the potential to suppress the release of Vpu-deficient HIV-1. However, the molecular mechanism(s) for tethering HIV-1 particles by hBST-2 remains unclear, and we speculated about the requirement for cellular co-factor(s) to trigger or assist its tethering ability. To explore this possibility, we utilize several cell lines derived from various species including human, AGM, dog, cat, rabbit, pig, mink, potoroo, and quail. We found that ectopic hBST-2 was efficiently expressed on the surface of all analyzed cells, and its expression suppressed the release of viral particles in a dose-dependent manner. These findings suggest that hBST-2 can tether HIV-1 particles without the need of additional co-factor(s) that may be expressed exclusively in primates, and thus, hBST-2 can also exert its function in many cells derived from a broad range of species. Interestingly, the suppressive effect of hBST-2 on HIV-1 release in Vero cells was much less pronounced than in the other examined cells despite the augmented surface expression of ectopic hBST-2 on Vero cells. Taken together, our findings suggest the existence of certain cell types in which hBST-2 cannot efficiently exert its inhibitory effect on virus release. The cell type-specific effect of hBST-2 may be critical to elucidate the mechanism of BST-2-dependent suppression of virus release

Functional mutations in spike glycoprotein of Zaire ebolavirus associated with an increase in infection efficiency

Ebola virus (EBOV) is extremely virulent, and its glycoprotein is necessary for viral entry. EBOV may adapt to its new host humans during outbreaks by acquiring mutations especially in glycoprotein, which allows EBOV to spread more efficiently. To identify these evolutionary selected mutations and examine their effects on viral infectivity, we used experimental–phylogenetic–structural interdisciplinary approaches. In evolutionary analysis of all available Zaire ebolavirus glycoprotein sequences, we detected two codon sites under positive selection, which are located near/within the region critical for the host‐viral membrane fusion, namely alanine‐to‐valine and threonine‐to‐isoleucine mutations at 82 (A82V) and 544 (T544I), respectively. The fine‐scale transmission dynamics of EBOV Makona variants that caused the 2014–2015 outbreak showed that A82V mutant was fixed in the population, whereas T544I was not. Furthermore, pseudotype assays for the Makona glycoprotein showed that the A82V mutation caused a small increase in viral infectivity compared with the T544I mutation. These findings suggest that mutation fixation in EBOV glycoprotein may be associated with their increased infectivity levels; the mutant with a moderate increase in infectivity will fix. Our findings showed that a driving force for Ebola virus evolution via glycoprotein may be a balance between costs and benefits of its virulence

Identification of the P-TEFb complex-interacting domain of Brd4 as an inhibitor of HIV-1 replication by functional cDNA library screening in MT-4 cells

AbstractWe conducted a phenotypic cDNA screening using a T cell line-based assay to identify human genes that render cells resistant to human immunodeficiency virus type 1 (HIV-1). We isolated potential HIV-1 resistance genes, including the carboxy terminal domain (CTD) of bromodomain-containing protein 4 (Brd4). Expression of GFP-Brd4-CTD was tolerated in MT-4 and Jurkat cells in which HIV-1 replication was markedly inhibited. We provide direct experimental data demonstrating that Brd4-CTD serves as a specific inhibitor of HIV-1 replication in T cells. Our method is a powerful tool for the identification of host factors that regulate HIV-1 replication in T cells

Sports Court-Based Camera Calibration Technique for Three-Dimensional Reconstruction of Knee Joint Kinematics

Videos of sports injuries provide beneficial insight for understanding their mechanisms. Recently, video analysis of actual injuries is particularly focused upon by researchers studying the anterior cruciate ligament（ ACL）. It is expected that establishing a technology capable of quantifying 3D knee kinematics using accidentally filmed videos of injuries could further our understanding of the ACL injury mechanism. The purpose of this study was to validate a camera calibration technique using sports court geometries to establish a methodology to extract 3D knee joint kinematics from uncalibrated video sequences. Four cameras were calibrated using line intersections and the vertical goal mouth of a handball court. A 1.6-m scaling pole, goniometerembedded link model, and jump-landing motions were filmed, and those 3D kinematics were reconstructed via the direct linear transformation method. The reconstructed values were compared with direct measurements and marker-based motion capture data to assess the accuracy of the reconstruction. Inter- and intra-operator differences were quantified to evaluate the objectivity and reliability of the technique. The length errors ranged from 0.65 to 1.7 cm and angular errors were 1.67°-3.90°. Knee flexion/extension was most accurately reconstructed（ errors: 2.71°-4.25°）, and the knee adduction/abduction showed moderate accuracy（ errors: 3.13°-5.35°）. The axial rotation showed large errors of 5.02°-6.53°. Although some limitations exist, the error ranges were small relative to those of previously reported knee displacements during actual ACL injury. Therefore, this technique can be used to quantify knee kinematics from uncalibrated video sequences with reasonable precision

Antithetic effect of interferon-α on cell-free and cell-to-cell HIV-1 infection

In HIV-1-infected individuals, transmitted/founder (TF) virus contributes to establish new infection and expands during the acute phase of infection, while chronic control (CC) virus emerges during the chronic phase of infection. TF viruses are more resistant to interferon-alpha (IFN-α)-mediated antiviral effects than CC virus, however, its virological relevance in infected individuals remains unclear. Here we perform an experimental-mathematical investigation and reveal that IFN-α strongly inhibits cell-to-cell infection by CC virus but only weakly affects that by TF virus. Surprisingly, IFN-α enhances cell-free infection of HIV-1, particularly that of CC virus, in a virus-cell density-dependent manner. We further demonstrate that LY6E, an IFN-stimulated gene, can contribute to the density-dependent enhancement of cell-free HIV-1 infection. Altogether, our findings suggest that the major difference between TF and CC viruses can be explained by their resistance to IFN-α-mediated inhibition of cell-to-cell infection and their sensitivity to IFN-α-mediated enhancement of cell-free infection

Multiomics Investigation Revealing the Characteristics of HIV-1-Infected Cells In Vivo

For eradication of HIV-1 infection, it is important to elucidate the detailed features and heterogeneity of HIV-1-infected cells in vivo. To reveal multiple characteristics of HIV-1-producing cells in vivo, we use a hematopoietic-stem-cell-transplanted humanized mouse model infected with GFP-encoding replication-competent HIV-1. We perform multiomics experiments using recently developed technology to identify the features of HIV-1-infected cells. Genome-wide HIV-1 integration-site analysis reveals that productive HIV-1 infection tends to occur in cells with viral integration into transcriptionally active genomic regions. Bulk transcriptome analysis reveals that a high level of viral mRNA is transcribed in HIV-1-infected cells. Moreover, single-cell transcriptome analysis shows the heterogeneity of HIV-1-infected cells, including CXCL13high cells and a subpopulation with low expression of interferon-stimulated genes, which can contribute to efficient viral spread in vivo. Our findings describe multiple characteristics of HIV-1-producing cells in vivo, which could provide clues for the development of an HIV-1 cure