ß-Coronaviruses Use Lysosomes for Egress Instead of the Biosynthetic Secretory Pathway.

ß-Coronaviruses are a family of positive-strand enveloped RNA viruses that includes the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Much is known regarding their cellular entry and replication pathways, but their mode of egress remains uncertain. Using imaging methodologies and virus-specific reporters, we demonstrate that ß-coronaviruses utilize lysosomal trafficking for egress rather than the biosynthetic secretory pathway more commonly used by other enveloped viruses. This unconventional egress is regulated by the Arf-like small GTPase Arl8b and can be blocked by the Rab7 GTPase competitive inhibitor CID1067700. Such non-lytic release of ß-coronaviruses results in lysosome deacidification, inactivation of lysosomal degradation enzymes, and disruption of antigen presentation pathways. ß-Coronavirus-induced exploitation of lysosomal organelles for egress provides insights into the cellular and immunological abnormalities observed in patients and suggests new therapeutic modalities.

Genetic Drivers of Epigenetic and Transcriptional Variation in Human Immune Cells.

Characterizing the multifaceted contribution of genetic and epigenetic factors to disease phenotypes is a major challenge in human genetics and medicine. We carried out high-resolution genetic, epigenetic, and transcriptomic profiling in three major human immune cell types (CD14+ monocytes, CD16+ neutrophils, and naive CD4+ T cells) from up to 197 individuals. We assess, quantitatively, the relative contribution of cis-genetic and epigenetic factors to transcription and evaluate their impact as potential sources of confounding in epigenome-wide association studies. Further, we characterize highly coordinated genetic effects on gene expression, methylation, and histone variation through quantitative trait locus (QTL) mapping and allele-specific (AS) analyses. Finally, we demonstrate colocalization of molecular trait QTLs at 345 unique immune disease loci. This expansive, high-resolution atlas of multi-omics changes yields insights into cell-type-specific correlation between diverse genomic inputs, more generalizable correlations between these inputs, and defines molecular events that may underpin complex disease risk.

Expanding the repertoire reveals recurrent, cryptic, and hematopoietic HLA class I minor histocompatibility antigens.

ABSTRACT: Allogeneic stem cell transplantation (alloSCT) is a curative treatment for hematological malignancies. After HLA-matched alloSCT, antitumor immunity is caused by donor T cells recognizing polymorphic peptides, designated minor histocompatibility antigens (MiHAs), that are presented by HLA on malignant patient cells. However, T cells often target MiHAs on healthy nonhematopoietic tissues of patients, thereby inducing side effects known as graft-versus-host disease. Here, we aimed to identify the dominant repertoire of HLA-I-restricted MiHAs to enable strategies to predict, monitor or modulate immune responses after alloSCT. To systematically identify novel MiHAs by genome-wide association screening, T-cell clones were isolated from 39 transplanted patients and tested for reactivity against 191 Epstein-Barr virus transformed B cell lines of the 1000 Genomes Project. By discovering 81 new MiHAs, we more than doubled the antigen repertoire to 159 MiHAs and demonstrated that, despite many genetic differences between patients and donors, often the same MiHAs are targeted in multiple patients. Furthermore, we showed that one quarter of the antigens are cryptic, that is translated from unconventional open reading frames, for example long noncoding RNAs, showing that these antigen types are relevant targets in natural immune responses. Finally, using single cell RNA-seq data, we analyzed tissue expression of MiHA-encoding genes to explore their potential role in clinical outcome, and characterized 11 new hematopoietic-restricted MiHAs as potential targets for immunotherapy. In conclusion, we expanded the repertoire of HLA-I-restricted MiHAs and identified recurrent, cryptic and hematopoietic-restricted antigens, which are fundamental to predict, follow or manipulate immune responses to improve clinical outcome after alloSCT.

H3N2 influenza A virus gradually adapts to human-type receptor binding and entry specificity after the start of the 1968 pandemic.

To become established upon zoonotic transfer, influenza A viruses (IAV) need to switch binding from "avian-type" α2-3-linked sialic acid receptors (2-3Sia) to "human-type" Siaα2-6-linked sialic acid receptors (2-6Sia). For the 1968 H3N2 pandemic virus, this was accomplished by two canonical amino acid substitutions in its hemagglutinin (HA) although a full specificity shift had not occurred. The receptor repertoire on epithelial cells is highly diverse and simultaneous interaction of a virus particle with a range of low- to very low-affinity receptors results in tight heteromultivalent binding. How this range of affinities determines binding selectivity and virus motility remains largely unknown as the analysis of low-affinity monovalent HA-receptor interactions is technically challenging. Here, a biolayer interferometry assay enabled a comprehensive analysis of receptor-binding kinetics evolution upon host-switching. Virus-binding kinetics of H3N2 virus isolates slowly evolved from 1968 to 1979 from mixed 2-3/2-6Sia specificity to high 2-6Sia specificity, surprisingly followed by a decline in selectivity after 1992. By using genetically tuned HEK293 cells, presenting either a simplified 2-3Sia- or 2-6Sia-specific receptor repertoire, receptor-specific binding was shown to correlate strongly with receptor-specific entry. In conclusion, the slow and continuous evolution of entry and receptor-binding specificity of seasonal H3N2 viruses contrasts with the paradigm that human IAVs need to rapidly acquire and maintain a high specificity for 2-6Sia. Analysis of the kinetic parameters of receptor binding provides a basis for understanding virus-binding specificity, motility, and HA/neuraminidase balance at the molecular level.

Kinetic analysis of paramyxovirus-sialoglycan receptor interactions reveals virion motility.

Many viruses initiate infection by binding to sialoglycan receptors at the cell surface. Binding to such receptors comes at a cost, however, as the sheer abundance of sialoglycans e.g. in mucus, may immobilize virions to non-functional decoy receptors. As a solution, sialoglycan-binding as well as sialoglycan-cleavage activities are often present in these viruses, which for paramyxoviruses are combined in the hemagglutinin-neuraminidase (HN) protein. The dynamic interactions of sialoglycan-binding paramyxoviruses with their receptors are thought to be key determinants of species tropism, replication and pathogenesis. Here we used biolayer interferometry to perform kinetic analyses of receptor interactions of animal and human paramyxoviruses (Newcastle disease virus, Sendai virus, and human parainfluenza virus 3). We show that these viruses display strikingly different receptor interaction dynamics, which correlated with their receptor-binding and -cleavage activities and the presence of a second sialic acid binding site. Virion binding was followed by sialidase-driven release, during which virions cleaved sialoglycans until a virus-specific density was reached, which was largely independent of virion concentration. Sialidase-driven virion release was furthermore shown to be a cooperative process and to be affected by pH. We propose that paramyxoviruses display sialidase-driven virion motility on a receptor-coated surface, until a threshold receptor density is reached at which virions start to dissociate. Similar motility has previously been observed for influenza viruses and is likely to also apply to sialoglycan-interacting embecoviruses. Analysis of the balance between receptor-binding and -cleavage increases our understanding of host species tropism determinants and zoonotic potential of viruses.

Neuraminidase-dependent entry of influenza A virus is determined by hemagglutinin receptor-binding specificity.

IMPORTANCE: Influenza A viruses (IAVs) contain hemagglutinin (HA) proteins involved in sialoglycan receptor binding and neuraminidase (NA) proteins that cleave sialic acids. While the importance of the NA protein in virion egress is well established, its role in virus entry remains to be fully elucidated. NA activity is needed for the release of virions from mucus decoy receptors, but conflicting results have been reported on the importance of NA activity in virus entry in the absence of decoy receptors. We now show that inhibition of NA activity affects virus entry depending on the receptor-binding properties of HA and the receptor repertoire present on cells. Inhibition of entry by the presence of mucus correlated with the importance of NA activity for virus entry, with the strongest inhibition being observed when mucus and OsC were combined. These results shed light on the importance in virus entry of the NA protein, an important antiviral drug target.

GPU-accelerated Bloch simulations and MR-STAT reconstructions using the Julia programming language.

PURPOSE: MR-STAT is a relatively new multiparametric quantitative MRI technique in which quantitative paramater maps are obtained by solving a large-scale nonlinear optimization problem. Managing reconstruction times is one of the main challenges of MR-STAT. In this work we leverage GPU hardware to reduce MR-STAT reconstruction times. A highly optimized, GPU-compatible Bloch simulation toolbox is developed as part of this work that can be utilized for other quantitative MRI techniques as well. METHODS: The Julia programming language was used to develop a flexible yet highly performant and GPU-compatible Bloch simulation toolbox called BlochSimulators.jl. The runtime performance of the toolbox is benchmarked against other Bloch simulation toolboxes. Furthermore, a (partially matrix-free) modification of a previously presented (matrix-free) MR-STAT reconstruction algorithm is proposed and implemented using the Julia language on GPU hardware. The proposed algorithm is combined with BlochSimulators.jl and the resulting MR-STAT reconstruction times on GPU hardware are compared to previously presented MR-STAT reconstruction times. RESULTS: The BlochSimulators.jl package demonstrates superior runtime performance on both CPU and GPU hardware when compared to other existing Bloch simulation toolboxes. The GPU-accelerated partially matrix-free MR-STAT reconstruction algorithm, which relies on BlochSimulators.jl, allows for reconstructions of 68 seconds per two-dimensional (2D slice). CONCLUSION: By combining the proposed Bloch simulation toolbox and the partially matrix-free reconstruction algorithm, 2D MR-STAT reconstructions can be performed in the order of one minute on a modern GPU card. The Bloch simulation toolbox can be utilized for other quantitative MRI techniques as well, for example for online dictionary generation for MR Fingerprinting.

High SNR full brain relaxometry at 7T by accelerated MR-STAT.

PURPOSE: To demonstrate the feasibility and robustness of the Magnetic Resonance Spin TomogrAphy in Time-domain (MR-STAT) framework for fast, high SNR relaxometry at 7T. METHODS: To deploy MR-STAT on 7T-systems, we designed optimized flip-angles using the BLAKJac-framework that incorporates the SAR-constraints. Transmit RF-inhomogeneities were mitigated by including a measured B 1 + $$ {B}_1^{+} $$ -map in the reconstruction. Experiments were performed on a gel-phantom and on five volunteers to explore the robustness of the sequence and its sensitivity to B 1 + $$ {B}_1^{+} $$ inhomogeneities. The SNR-gain at 7T was explored by comparing phantom and in vivo results to MR-STAT at 3T in terms of SNR-efficiency. RESULTS: The higher SNR at 7T enabled two-fold acceleration with respect to current 2D MR-STAT protocols at lower field strengths. The resulting scan had whole-brain coverage, with 1 x 1 x 3 mm3 resolution (1.5 mm slice-gap) and was acquired within 3 min including the B 1 + $$ {B}_1^{+} $$ -mapping. After B 1 + $$ {B}_1^{+} $$ -correction, the estimated T1 and T2 in a phantom showed a mean relative error of, respectively, 1.7% and 4.4%. In vivo, the estimated T1 and T2 in gray and white matter corresponded to the range of values reported in literature with a variation over the subjects of 1.0%-2.1% (WM-GM) for T1 and 4.3%-5.3% (WM-GM) for T2. We measured a higher SNR-efficiency at 7T (R = 2) than at 3T for both T1 and T2 with, respectively, a 4.1 and 2.3 times increase in SNR-efficiency. CONCLUSION: We presented an accelerated version of MR-STAT tailored to high field (7T) MRI using a low-SAR flip-angle train and showed high quality parameter maps with an increased SNR-efficiency compared to MR-STAT at 3T.

An improved method to measure transfer functions using MRI.

PURPOSE: A previously published method for MRI-based transfer function assessment makes use of the so-called transceive phase assumption (TPA). This limits its applicability to shorter leads and/or lower field strengths. A new method is presented where the background electric field is determined from both B 1 + $$ {\mathrm{B}}_1^{+} $$ - and B 1 - $$ {\mathrm{B}}_1^{-} $$ -field distributions, avoiding the TPA and making it more generally applicable. THEORY AND METHODS: These B 1 $$ {\mathrm{B}}_1 $$ -distributions are determined from a spoiled gradient echo multiflip angle acquisition. From the separated B 1 $$ {\mathrm{B}}_1 $$ -components the background electrical field and the induced current are computed. Further improvement is achieved by recasting the B 1 $$ {\mathrm{B}}_1 $$ -field model as a "magnitude squared least squares" problem. The proposed reconstruction method is used to determine transfer functions of various copper wire lengths up to 40 cm inside an elliptical ASTM phantom. The method is first tested on EM-simulated data and subsequently phantom and bench measurements are used to determine transfer functions experimentally. RESULTS: In silica reconstructions demonstrate the validity of the proposed B 1 $$ {\mathrm{B}}_1 $$ -field model resulting in highly accurate reconstructed B 1 $$ {\mathrm{B}}_1 $$ -fields, currents, incident electric fields and transfer functions. The experimental results show slight deviations in the field model, however, resulting transfer functions are accurately determined with high similarity to simulations and comparable to bench measurements. CONCLUSION: A more generally applicable method for MRI-based transfer function assessment is presented. The proposed method circumvents phase assumptions making it applicable for longer objects and/or higher field strengths. Additional improvements are implemented in the B 1 $$ {\mathrm{B}}_1 $$ -mapping method and the solution algorithm.

Water diffusion and T2 quantification in transient-state MRI: the effect of RF pulse sequence.

In quantitative measurement of the T 2 value of tissues, the diffusion of water molecules has been recognized as a confounder. This is most notably so for transient-state quantitative mapping techniques, which allow simultaneous estimation of T 1 and T 2 . In prior work, apparently conflicting conclusions are presented on the level of diffusion-induced bias on the T2 estimate. So far there is a lack of studies on the effect of the RF pulse angle sequence on the level of diffusion-induced bias. In this work, we show that the specific transient-state RF pulse sequence has a large effect on this level of bias. In particular, the bias level is strongly influenced by the mean value of the RF pulse angles. Also, for realistic values of the spoiling gradient area, we infer that the diffusion-induced bias is negligible for non-liquid human tissues; yet, for phantoms, the effect can be substantial (15% of the true T 2 value) for some RF pulse sequences. This should be taken into account in validation procedures.

A three-dimensional Magnetic Resonance Spin Tomography in Time-domain protocol for high-resolution multiparametric quantitative magnetic resonance imaging.

Magnetic Resonance Spin TomogrAphy in Time-domain (MR-STAT) is a multiparametric quantitative MR framework, which allows for simultaneously acquiring quantitative tissue parameters such as T1, T2, and proton density from one single short scan. A typical two-dimensional (2D) MR-STAT acquisition uses a gradient-spoiled, gradient-echo sequence with a slowly varying RF flip-angle train and Cartesian readouts, and the quantitative tissue maps are reconstructed by an iterative, model-based optimization algorithm. In this work, we design a three-dimensional (3D) MR-STAT framework based on previous 2D work, in order to achieve better image signal-to-noise ratio, higher though-plane resolution, and better tissue characterization. Specifically, we design a 7-min, high-resolution 3D MR-STAT sequence, and the corresponding two-step reconstruction algorithm for the large-scale dataset. To reduce the long acquisition time, Cartesian undersampling strategies such as SENSE are adopted in our transient-state quantitative framework. To reduce the computational burden, a data-splitting scheme is designed for decoupling the 3D reconstruction problem into independent 2D reconstructions. The proposed 3D framework is validated by numerical simulations, phantom experiments, and in vivo experiments. High-quality knee quantitative maps with 0.8 × 0.8 × 1.5 mm3 resolution and bilateral lower leg maps with 1.6 mm isotropic resolution can be acquired using the proposed 7-min acquisition sequence and the 3-min-per-slice decoupled reconstruction algorithm. The proposed 3D MR-STAT framework could have wide clinical applications in the future.

Autonomous B-cell receptor signaling and genetic aberrations in chronic lymphocytic leukemia-phenotype monoclonal B lymphocytosis in siblings of patients with chronic lymphocytic leukemia.

Clonal expansion of CD5-expressing B cells, commonly designated as monoclonal B lymphocytosis (MBL), is a precursor condition for chronic lymphocytic leukemia (CLL). The mechanisms driving subclinical MBL B-cell expansion and progression to CLL, occurring in approximately 1% of affected individuals, are unknown. An autonomously signaling B-cell receptor (BCR) is essential for the pathogenesis of CLL. The objectives of this study were functional characterization of the BCR of MBL in siblings of CLL patients and a comparison of genetic variants in MBL-CLL sibling pairs. Screening of peripheral blood by flow cytometry detected 0.2-480 clonal CLL-phenotype cells per microliter (median: 37/µL) in 34 of 191 (17.8%) siblings of CLL patients. Clonal BCR isolated from highly purified CLL-phenotype cells induced robust calcium mobilization in BCR-deficient murine pre-B cells in the absence of external antigen and without experimental crosslinking. This autonomous BCR signal was less intense than the signal originating from the CLL BCR of their CLL siblings. According to genotyping by single nucleotide polymorphism array, whole exome, and targeted panel sequencing, CLL risk alleles were found with high and similar prevalence in CLL patients and MBL siblings, respectively. Likewise, the prevalence of recurrent CLL-associated genetic variants was similar between CLL and matched MBL samples. However, copy number variations and small variants were frequently subclonal in MBL cells, suggesting their acquisition during subclinical clonal expansion. These findings support a stepwise model of CLL pathogenesis, in which autonomous BCR signaling leads to a non-malignant (oligo)clonal expansion of CD5+ B cells, followed by malignant progression to CLL after acquisition of pathogenic genetic variants.

Trophic Transfer and Toxic Potency of Rare Earth Elements along a Terrestrial Plant-Herbivore Food Chain.

Extensive rare earth element (REE) mining activities have caused REE contamination of ambient agricultural soils, posing threats to associated food webs. Here, a simulated lettuce-snail food chain was conducted to evaluate the trophic transfer characteristics and the consequent effects of REEs on consumers. After 50-day exposure to soil, lettuce roots dose-dependently accumulated 9.4-76 mg kg-1 REEs and translocated 3.7-20 mg kg-1 REEs to shoots. Snails feeding on REE-contaminated shoots accumulated 3.0-6.7 mg kg-1 REEs with trophic transfer factors of 0.20-0.98, indicating trophic dilution in the lettuce-snail system. REE profiles in lettuce and snails indicated light REE (LREE) enrichment only in snails and the varied REE profiles along the food chain. This was corroborated by toxicokinetics. Estimated uptake (Ku) and elimination (Ke) parameters were 0.010-2.9 kgshoot kgsnail-1 day-1 and 0.010-1.8 day-1, respectively, with higher Ku values for LREE and HREE. The relatively high Ke, compared to Ku, indicating a fast REE elimination, supports the trophic dilution. Dietary exposure to REEs dose-dependently affected gut microbiota and metabolites in snails. These effects are mainly related to oxidative damage and energy expenditure, which are further substantiated by targeted analysis. Our study provides essential information about REE bioaccumulation characteristics and its associated risks to terrestrial food chains near REE mining areas.

Two separate mechanisms are involved in membrane permeabilization during lipid oxidation.

Lipid oxidation is a universal degradative process of cell membrane lipids that is induced by oxidative stress and reactive oxygen and nitrogen species (RONS) in multiple pathophysiological situations. It has been shown that certain oxidized lipids alter membrane properties, leading to a loss of membrane function. Alteration of membrane properties is thought to depend on the initial membrane lipid composition, such as the number of acyl chain unsaturations. However, it is unclear how oxidative damage is related to biophysical properties of membranes. We therefore set out to quantify lipid oxidation through various analytical methods and determine key biophysical membrane parameters using model membranes containing lipids with different degrees of lipid unsaturation. As source for RONS, we used cold plasma, which is currently developed as treatment for infections and cancer. Our data revealed complex lipid oxidation that can lead to two main permeabilization mechanisms. The first one appears upon direct contact of membranes with RONS and depends on the formation of truncated oxidized phospholipids. These lipids seem to be partly released from the bilayer, implying that they are likely to interact with other membranes and potentially act as signaling molecules. This mechanism is independent of lipid unsaturation, does not rely on large variations in lipid packing, and is most probably mediated via short-living RONS. The second mechanism takes over after longer incubation periods and probably depends on the continued formation of lipid oxygen adducts such as lipid hydroperoxides or ketones. This mechanism depends on lipid unsaturation and involves large variations in lipid packing. This study indicates that polyunsaturated lipids, which are present in mammalian membranes rather than in bacteria, do not sensitize membranes to instant permeabilization by RONS but could promote long-term damage.

Data-driven electrical conductivity brain imaging using 3 T MRI.

Magnetic resonance electrical properties tomography (MR-EPT) is a non-invasive measurement technique that derives the electrical properties (EPs, e.g., conductivity or permittivity) of tissues in the radiofrequency range (64 MHz for 1.5 T and 128 MHz for 3 T MR systems). Clinical studies have shown the potential of tissue conductivity as a biomarker. To date, model-based conductivity reconstructions rely on numerical assumptions and approximations, leading to inaccuracies in the reconstructed maps. To address such limitations, we propose an artificial neural network (ANN)-based non-linear conductivity estimator trained on simulated data for conductivity brain imaging. Network training was performed on 201 synthesized T2-weighted spin-echo (SE) data obtained from the finite-difference time-domain (FDTD) electromagnetic (EM) simulation. The dataset was composed of an approximated T2-w SE magnitude and transceive phase information. The proposed method was tested three in-silico and in-vivo on two volunteers and three patients' data. For comparison purposes, various conventional phase-based EPT reconstruction methods were used that ignore B 1 + magnitude information, such as Savitzky-Golay kernel combined with Gaussian filter (S-G Kernel), phase-based convection-reaction EPT (cr-EPT), magnitude-weighted polynomial-fitting phase-based EPT (Poly-Fit), and integral-based phase-based EPT (Integral-based). From the in-silico experiments, quantitative analysis showed that the proposed method provides more accurate and improved quality (e.g., high structural preservation) conductivity maps compared to conventional reconstruction methods. Representatively, in the healthy brain in-silico phantom experiment, the proposed method yielded mean conductivity values of 1.97 ± 0.20 S/m for CSF, 0.33 ± 0.04 S/m for WM, and 0.52 ± 0.08 S/m for GM, which were closer to the ground-truth conductivity (2.00, 0.30, 0.50 S/m) than the integral-based method (2.56 ± 2.31, 0.39 ± 0.12, 0.68 ± 0.33 S/m). In-vivo ANN-based conductivity reconstructions were also of improved quality compared to conventional reconstructions and demonstrated network generalizability and robustness to in-vivo data and pathologies. The reported in-vivo brain conductivity values were in agreement with literatures. In addition, the proposed method was observed for various SNR levels (SNR levels = 10, 20, 40, and 58) and repeatability conditions (the eight acquisitions with the number of signal averages = 1). The preliminary investigations on brain tumor patient datasets suggest that the network trained on simulated dataset can generalize to unforeseen in-vivo pathologies, thus demonstrating its potential for clinical applications.

Varying Viral Replication and Disease Profiles of H2N2 Influenza in Ferrets Is Associated with Virus Isolate and Inoculation Route.

H2N2 influenza virus, the causative agent of the 1957 "Asian flu" pandemic, has disappeared from circulation. However, H2-influenza viruses are still circulating in avian reservoirs. Combined with the waning of H2N2-specific immunity in the human population, there is a risk of reintroduction of H2N2 influenza virus. Vaccines could help in preventing a future pandemic, but to assess their efficacy animal models are required. We therefore set out to expand the ferret model for H2N2 influenza disease by infecting ferrets intranasally or intratracheally with four different H2N2 viruses to investigate their influence on the severity of disease. The H2N2 viruses were collected either during the pandemic or near the end of H2N2 circulation and covered both clade I and clade II viruses. Infection of ferrets with the different viruses showed that viral replication, disease, and pathology differed markedly between virus isolates and infection routes. Intranasal inoculation induced a severe to mild rhinitis, depending on the virus isolate, and did not lead to lung infection or pathology. When administered intratracheally, isolates that successfully replicated in the lower respiratory tract (LRT) induced a nonlethal disease that resembles that of a moderate pneumonia in humans. Differences in viral replication and disease between viruses could be associated with their binding preference for α2,3- and α2,6-sialic acid. The model presented here could facilitate the development of a new generation of H2N2 influenza vaccines. IMPORTANCE In 1957 the world was subjected to a pandemic caused by an influenza A virus of the subtype H2N2. Although the virus disappeared in 1968, H2 viruses continue to circulate in avian reservoirs. It is therefore possible that the H2N2 influenza virus will be reintroduced into the human population, which can lead to another pandemic. The impact of a new H2N2 influenza pandemic can be mitigated by vaccination. However, these vaccines first need to be developed and tested in animal models. In preparation for this, we expanded the ferret model to mimic the different facets of human H2N2 influenza infection and disease. This model can be used for the development and evaluation of new H2N2 influenza vaccines.

Difference in respiratory syncytial virus-specific Fc-mediated antibody effector functions between children and adults.

Respiratory syncytial virus (RSV) infections are a major cause of bronchiolitis and pneumonia in infants and older adults, for which there is no known correlate of protection. Increasing evidence suggests that Fc-mediated antibody effector functions have an important role, but little is known about the development, heterogeneity, and durability of these functional responses. In light of future vaccine strategies, a clear view of the immunological background and differences between various target populations is of crucial importance. In this study, we have assessed both quantitative and qualitative aspects of RSV-specific serum antibodies, including IgG/IgA levels, IgG subclasses, antibody-dependent complement deposition, cellular phagocytosis, and NK cell activation (ADNKA). Samples were collected cross-sectionally in different age groups (11-, 24-, and 46-month-old children, adults, and older adults; n = 31-35 per group) and longitudinally following natural RSV infection in (older) adults (2-36 months post-infection; n = 10). We found that serum of 24-month-old children induces significantly lower ADNKA than the serum of adults (P < 0.01), which is not explained by antibody levels. Furthermore, in (older) adults we observed boosting of antibody levels and functionality at 2-3 months after RSV infection, except for ADNKA. The strongest decrease was subsequently observed within the first 9 months, after which levels remained relatively stable up to three years post-infection. Together, these data provide a comprehensive overview of the functional landscape of RSV-specific serum antibodies in the human population, highlighting that while antibodies reach adult levels already at a young age, ADNKA requires more time to fully develop.

Multi-echo MR thermometry in the upper leg at 7 T using near-harmonic 2D reconstruction for initialization.

PURPOSE: The aim of this work is the development of a thermometry method to measure temperature increases in vivo, with a precision and accuracy sufficient for validation against thermal simulations. Such an MR thermometry model would be a valuable tool to get an indication on one of the major safety concerns in MR imaging: the tissue heating occurring due to radiofrequency (RF) exposure. To prevent excessive temperature rise, RF power deposition, expressed as specific absorption rate, cannot exceed predefined thresholds. Using these thresholds, MRI has demonstrated an extensive history of safe usage. Nevertheless, MR thermometry would be a valuable tool to address some of the unmet needs in the area of RF safety assessment, such as validation of specific absorption rate and thermal simulations, investigation of local peak temperatures during scanning, or temperature-based safety guidelines. METHODS: The harmonic initialized model-based multi-echo approach is proposed. The method combines a previously published model-based multi-echo water/fat separated approach with an also previously published near-harmonic 2D reconstruction method. The method is tested on the human thigh with a multi-transmit array at 7 T, in three volunteers, and for several RF shims. RESULTS: Precision and accuracy are improved considerably compared to a previous fat-referenced method (precision: 0.09 vs. 0.19°C). Comparison of measured temperature rise distributions to subject-specific simulated counterparts show good relative agreement for multiple RF shim settings. CONCLUSION: The high precision shows promising potential for validation purposes and other RF safety applications.

Efficient performance analysis and optimization of transient-state sequences for multiparametric magnetic resonance imaging.

In transient-state multiparametric MRI sequences such as Magnetic Resonance Spin TomogrAphy in Time-domain (MR-STAT), MR fingerprinting, or hybrid-state imaging, the flip angle pattern of the RF excitation varies over the sequence. This gives considerable freedom to choose an optimal pattern of flip angles. For pragmatic reasons, most optimization methodologies choose for a single-voxel approach (i.e., without taking the spatial encoding scheme into account). Particularly in MR-STAT, the context of spatial encoding is important. In the current study, we present a methodology, called BLock Analysis of a K-space-domain Jacobian (BLAKJac), which is sufficiently fast to optimize a sequence in the context of a predetermined phase-encoding pattern. Based on MR-STAT acquisitions and reconstructions, we show that sequences optimized using BLAKJac are more reliable in terms of actually achieved precision than conventional single-voxel-optimized sequences. In addition, BLAKJac provides analytical tools that give insights into the performance of the sequence in a very limited computation time. Our experiments are based on MR-STAT, but the theory is equally valid for other transient-state multiparametric methods.

Tier-based formalism for safety assessment of custom-built radio-frequency transmit coils.

The purpose of this work is to propose a tier-based formalism for safety assessment of custom-built radio-frequency (RF) coils that balances validation effort with the effort put in determinating the safety factor. The formalism has three tier levels. Higher tiers require increased effort when validating electromagnetic simulation results but allow for less conservative safety factors. In addition, we propose a new method to calculate modeling uncertainty between simulations and measurements and a new method to propagate uncertainties in the simulation into a safety factor that minimizes the risk of underestimating the peak specific absorption rate (SAR). The new safety assessment procedure was completed for all tier levels for an eight-channel dipole array for prostate imaging at 7 T and an eight-channel dipole array for head imaging at 10.5 T, using data from two different research sites. For the 7 T body array, the validation procedure resulted in a modeling uncertainty of 77% between measured and simulated local SAR distributions. For a situation where RF shimming is performed on the prostate, average power limits of 2.4 and 4.5 W/channel were found for tiers 2 and 3, respectively. When the worst-case peak SAR among all phase settings was calculated, power limits of 1.4 and 2.7 W/channel were found for tiers 2 and 3, respectively. For the 10.5 T head array, a modeling uncertainty of 21% was found based on B1 + mapping. For the tier 2 validation, a power limit of 2.6 W/channel was calculated. The demonstrated tier system provides a strategy for evaluating modeling inaccuracy, allowing for the rapid translation of novel coil designs with conservative safety factors and the implementation of less conservative safety factors for frequently used coil arrays at the expense of increased validation effort.