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Jimenez, A.

Publications and source records attributed to Jimenez, A..

At least 19 recordsLinked to original sources

Coordinated Brain-Network Dynamics of Maternal Adaptation

The maternal brain exhibits impressive plasticity, undergoing many anatomical and functional alterations over the transition to parenting. Such alterations can reflect maternal adaptation to offspring needs, aligning with the onset and maintenance of maternal care behaviors. These brain-wide neural changes occur across stages, starting during pregnancy and continuing through the peripartal transition. We therefore wanted to understand brain changes with pregnancy, parenting, and care behaviors over time. Many brain regions have been implicated in the coordination of maternal adaptation and corresponding care behaviors. We used computational modeling of multi-region neural oscillation data to identify distinct electrical networks during naturally-occurring care behaviors and across multiple maternal stages in outbred, highly maternal mice. This work demonstrates that multi-region oscillatory electrical dynamics at high spatiotemporal resolution reflect these functions. Distinct, discriminative networks predict maternal behavior and stage, showcasing network adaptation with parental transition. Maternal behavior and stage also had substantial effects on other distinct, multi-region oscillatory electrical networks relevant to stress susceptibility, further linking maternal adaptation to the neurophysiology of stress. While these stress networks were perturbed by maternal experience of early life stress, networks trained on maternal behavior and stage remained robust against such perturbation. This suggests a uniquely persistent strength of maternal electrical dynamics. Together, this work supports the role of oscillatory electrical dynamics as networks underlying fundamentally conserved behaviors and transitions, demonstrating wide relevance and significance to brain-behavior discovery.

neuroscience↗

Robust mammalian RNA localization elements are complex and multipartite

The subcellular localization patterns of RNAs are controlled by regulatory elements contained within them. However, for most localized RNAs, the identities of these elements remain unknown. We had previously identified several localization elements that are necessary and sufficient for robust, kinesin-dependent RNA targeting to microtubule plus ends in a variety of cell types. Yet the characteristics of these elements that are critical for function remained unclear. To address this, we systematically created tens of thousands of mutant localization elements and quantified their ability to regulate subcellular RNA localization in neuronal cells. We found that the minimally active size of these localization elements is large, approximately 200 nucleotides. These elements contain multiple important subsequences, with some being completely intolerant of any changes and others being tolerant to a shuffling of nucleotide order but not to changes in nucleotide composition. Using single molecule microscopy, we verified these findings in primary rat neurons. Together, these results demonstrate that highly active mammalian RNA localization elements are large, complex, and multipartite and lay a foundation for further mechanistic studies of their function.

molecular biology↗

BCG-induced reprogramming of monocyte/macrophage populations enhances lung antitumor immunity in mice

The tumor microenvironment (TME) significantly influences antitumor immunity, with monocytes and macrophages playing pivotal roles both in pro- and anti-tumoral functions. Tumor-associated macrophages (TAMs) often adopt immunosuppressive phenotypes that promote tumor progression by inhibiting cytotoxic T and NK cells. This study investigates the antitumor mechanisms of intravenous (IV) Bacillus Calmette-Guerin (BCG) in a B16-F10 lung melanoma mouse model, focusing on its impact on monocyte/macrophage populations. Single-cell RNA sequencing revealed that IV BCG reprograms tumor-associated monocyte-derived macrophages (mo-macs), shifting them from immunosuppressive to pro-inflammatory phenotypes enriched in interferon-response signatures. BCG treatment increased the recruitment of classical (Mon Iigp1) and non-classical (Mon Fcgr4) monocytes, which exhibited enhanced antigen presentation and pro-inflammatory cytokine production, while reducing immunosuppressive subsets prevalent in untreated controls. These BCG-induced mo-macs established robust interactions with NK and T cells, promoting their activation and enhancing cytotoxic function, as validated by functional assays. Notably, transfer of BCG reprogrammed bone marrow progenitors into naive recipients elicited a sustained generation of immunostimulatory mo-macs that enhanced NK and T cell responses upon tumor challenge, These findings highlight IV BCGs potential as a cancer immunotherapy that targets the myeloid compartment to foster a pro-inflammatory TME, offering durable antitumor immunity by engaging both innate and adaptive immune responses.

immunology↗

Harmonic Amplitude-Modulated Singular Value Decomposition for Ultrafast Ultrasound Imaging of Gas Vesicles

Ultrafast nonlinear ultrasound imaging of gas vesicles (GV) contrast agents promises high-sensitivity biomolecular visualization with applications such as targeted molecular imaging of tumor markers or real-time tracking of gene expression. However, separating GV-specific signal from tissue remains challenging and requires the implementation of complex transmit schemes. In this work we introduce harmonic amplitude-modulated singular value decomposition (HAM-SVD), which synergizes pulse inversion (PI) with amplitude-modulated singular value decomposition (AM-SVD) to isolate GV-specific second-harmonic signals. In HAM-SVD, single-cycle plane waves at 9.6 MHz and five tilted angles (at a pulse repetition frequency of 2500 Hz) are transmitted under four duty cycles with alternating polarity. Beamformed IQ data are reshaped along a "space x pressure" matrix and decomposed via SVD; tissue background is cancelled by discarding the first and lowest singular modes, yielding an image comprised solely of pressure-dependent second harmonic signals. HAM-SVD sequence enables wide-field, ultrafast imaging without complex transmit sequences. Validation via simulations, in vitro phantoms, and in vivo rat lower limb experiments demonstrates HAM-SVDs outperformance compared to PI and AM-SVD. HAM-SVD is shown to achieve a 19.16 {+/-} 1.63 dB signal-to-background ratio (SBR) in vivo, surpassing PI (14.19 {+/-} 1.41 dB) and AM-SVD (15.79 {+/-} 1.38 dB). HAM-SVD overcomes limitations of conventional nonlinear techniques (e.g., depth restrictions, tissue clutter) by combining PIs harmonic sensitivity with AM-SVDs adaptive clutter filtering of tissue signals. This approach enhances molecular imaging specificity for GVs and holds potential for ultrasound localization microscopy of slow-flowing agents.

bioengineering↗

Contributions of Folded and Disordered Domains to RNA Binding by HNRNPR

RNA binding proteins (RBPs) interact with and tightly regulate the fate of messenger RNAs but how RNA targets are recognized remains a challenging question. RBPs often contain multiple domains known to directly bind RNA, such as RNA recognition motifs (RRMs), as well as domains whose RNA binding capacity remains incompletely understood, e.g., low complexity domains (LCDs). Here, we dissect HNRNPR, an RBP with three RRMs and an arginine-glycine rich (RG-rich) LCD. We apply unbiased high-throughput biochemical approaches and identify critical RNA binding domains that confer specificity. We show that not all RRMs contribute equally to binding and find that RRM3, along with a downstream C-terminal charged region, are required for RNA binding. We find that HNRNPR also binds RNA G-quadruplexes (rG4s) and map multiple rG4 binding sites including RRM3 with the C-terminal charged region and RG-rich regions within the LCD. We dissect rG4 specificity for the full length HNRNPR and LCD using a newly created RNA pool focused on rG4s and reveal that binding is dependent on RNA folding and find specific rG4 features that enhance HNRNPR-rG4 interactions. Our work highlights the complexity of RBP-RNA interactions and motivates the study of disordered regions as RNA binding domains.

biochemistry↗

Polyglutamine expansion induced dynamic misfolding of Androgen Receptor

Spinal bulbar muscular atrophy (SBMA) is caused by a polyglutamine expansion (pQe) in the N-terminal transactivation domain of human androgen receptor (AR-NTD), resulting in a combination of toxic gain- and loss-of-function mechanisms. The structural basis of these processes has not been resolved due to the disordered nature of the NTD, which hinders experimental analyses of its detailed conformations. Here, using extensive computational modelling, we show that AR-NTD forms dynamic compact regions, which upon pQe re-organize dynamically, mediated partly by direct pQ interaction with the Androgen N-Terminal Signature (ANTS) motif. The altered dynamics of the NTD result in a perturbation of interdomain interactions, with potential implications for binding of the receptor protein to its response element. Oligomeric aggregation of the dynamic misfolded NTD exposes pQe, but blocks tau-5 and the FQNLF motif, which could lead to aberrant receptor transcriptional activity. These observations suggest a structural mechanism for AR dysfunction in SBMA.

pathology↗

Peripheral CGRP engages brain-wide electrical network activity of migraine

BackgroundMigraine is a disorder of severe, recurrent headaches and debilitating sensory, cognitive and affective symptoms, often triggered by stress. Early life stress in childhood has been shown to increase the likelihood of migraine in adulthood in humans. Calcitonin-gene relate peptide (CGRP) has been shown to reliably and acutely induce migraine or migraine-like behavior in both humans and rodent models. Here we investigate the impact of early life stress and CGRP on migraine-related neural circuitry, as well as the impact of early life stress on CGRP-mediated migraine-like activity in order to better understand the mechanisms by which early life stress predisposes neural circuitry to migraine brain activity. MethodsWe implemented an early life stress paradigm in the outbred strain of mice, CD1. We evaluated the impact of peripheral CGRP on migraine-like behavior and employed multi-site in vivo neurophysiology in freely behaving mice. A changepoint analysis was used to dissect differences in individual CGRP-induced responses. ResultsWe found that early life stress exacerbated migraine-related behavioral and network physiology. CGRP alone caused disruptions in neural oscillatory activity across a network of brain regions including the anterior cingulate cortex (ACC), amygdala (AMY), thalamus (Po, VPM, and MDthal), and parabrachial nucleus (PBN). We found that power across the network was lowered within 10 minutes of peripheral CGRP exposure, which was sustained for [~]40-50 min. Coherence was mostly disrupted in amygdalar brain region pairings, and took on a shorter timecourse, with partial rescue of these responses by migraine abortive, sumatriptan. We found that early life stress exacerbated most of these responses, especially AMY-thalamic coherence pairings, although early life stress in the absence of CGRP demonstrated no impact on the network overall. We further identified individual mice with brain-network activity hypersusceptible to migraine. ConclusionsOur findings demonstrate that early life stress confers vulnerability to migraine, simultaneously impacting behavior and brain network activity responses to peripheral CGRP.

neuroscience↗

Darwin's Naturalization Conundrum explained by gradients of environmental stress and disturbance

Darwins Naturalization Conundrum (DNC) states that non-native species closely related to the native community are either more likely to succeed because shared adaptations help them overcome environmental filtering, or less likely to succeed because of strong competition with their native relatives. Despite extensive research, no general patterns have so far emerged. One reason may be that the relative importance of competition and environmental filtering depends on environmental conditions. To test this hypothesis, we conducted a global assessment of DNC examining patterns of phylogenetic relatedness of non-native plant species to the native community along gradients of elevation and anthropogenic disturbance in mountains. Phylogenetic distance of non-native to native species decreased with increasing elevation and in disturbed plant communities. Our results help resolve DNC by showing that the environmental context sets expectations for patterns of relatedness between non-native and native species and helps illuminate the ecological and evolutionary processes generating these patterns.

ecology↗

Nonlinear Singular Value Decomposition Beamforming for Ultrasound Imaging of Gas Vesicles

Ultrasound imaging holds significant promise for the observation of molecular and cellular phenomena through the utilization of acoustic contrast agents and acoustic reporter genes. Optimizing imaging methodologies for enhanced detection represents an imperative advancement in this field. Most advanced techniques relying on amplitude modulation scheme such as cross amplitude modulation (xAM) and ultrafast amplitude modulation (uAM) combined with Hadamard encoded multiplane wave transmissions have shown efficacy in capturing acoustic signals of gas vesicles (GVs). Nonetheless, uAM sequence requires odd- or even-element transmissions leading to imprecise amplitude modulation emitting scheme, and the complex multiplane wave transmission scheme inherently yields overlong pulse durations. xAM sequence is limited in terms of field of view and imaging depth. To overcome these limitations, we introduce an innovative ultrafast imaging sequence called nonlinear singular value decomposition (SVD) beamforming. Our method demonstrated a contrast imaging sensitivity comparable to the current gold-standard xAM and uAM, while requiring 4.8 times less pulse transmissions. With similar number of transmit pulses, nonlinear SVD beamforming outperforms xAM and uAM in terms of an improvement in signal-to-background ratio of + 4.78 {+/-} 0.35 dB and + 8.29 {+/-} 3.52 dB respectively. Additionally, our method provides a higher flexibility in terms of the selection of acoustic pressure amplitude compared to the other methods. Furthermore, it shows a significant potential for application in the realm of ultrasound localization microscopy (ULM), where it stands poised to facilitate the more precise extraction of nonlinear signatures originating from contrast agents.

biophysics↗

G4mer: An RNA language model for transcriptome-wide identification of G-quadruplexes and disease variants from population-scale genetic data

RNA G-quadruplexes (rG4s) are key regulatory elements in gene expression, yet the effects of genetic variants on rG4 formation remain underexplored. Here, we introduce G4mer, an RNA language model that predicts rG4 formation and evaluates the effects of genetic variants across the transcriptome. G4mer significantly improves accuracy over existing methods, highlighting sequence length and flanking motifs as important rG4 features. Applying G4mer to 5 untranslated region (UTR) variations, we identify variants in breast cancer-associated genes that alter rG4 formation and validate their impact on structure and gene expression. These results demonstrate the potential of integrating computational models with experimental approaches to study rG4 function, especially in diseases where non-coding variants are often overlooked. To support broader applications, G4mer is available as both a web tool and a downloadable model.

genetics↗

Murine modeling of menstruation identifies immune correlates of protection during Chlamydia muridarum challenge.

The menstrual cycle influences the risk of acquiring sexually transmitted infections (STIs), including Chlamydia trachomatis (C. trachomatis), although the underlying immune contributions are poorly defined. A mouse model simulating the immune-mediated process of menstruation could provide valuable insights into tissue-specific determinants of protection against chlamydial infection within the cervicovaginal and uterine mucosae comprising the female reproductive tract (FRT). Here, we used the pseudopregnancy approach in naive C57Bl/6 mice and performed vaginal challenge with Chlamydia muridarum (C. muridarum) at decidualization, endometrial tissue remodeling, or uterine repair. This strategy identified that the time frame comprising uterine repair correlated with robust infection and greater bacterial burden as compared with mice on hormonal contraception, while challenges during endometrial remodeling were least likely to result in a productive infection. By comparing the infection site at early time points following chlamydial challenge, we found that a greater abundance of innate effector populations and proinflammatory signaling, including IFN{psi} correlated with protection. FRT immune profiling in uninfected mice over pseudopregnancy or in pig-tailed macaques over the menstrual cycle identified NK cell infiltration into the cervicovaginal tissues and lumen over the course of endometrial remodeling. Notably, NK cell depletion over this time frame reversed protection, with mice now productively infected with C. muridarum following challenge. This study shows that the pseudopregnancy murine menstruation model recapitulates immune changes in the FRT as a result of endometrial remodeling and identifies NK cell localization at the FRT as essential for immune protection against primary C. muridarum infection. Author SummaryAlthough the vast majority of women and adolescent girls of reproductive age experience menstruation, we have little insight into how this tissue remodeling process alters mucosal immune defenses against infection by genitourinary pathogens. In this study, we used a murine model of menstruation to investigate how endometrial shedding and repair alters the immune landscape in the female reproductive tract (FRT) to influence chlamydial infections. Using this approach, we identified that endometrial remodeling regulates a substantial pro-inflammatory immune response, including NK cell recruitment into the cervicovaginal tissues, and we further confirmed this phenomenon is occurring in a naturally menstruating species. The localization of NK cells in the FRT at the time of challenge was determined to be responsible for rapid immune protection that reduced C. muridarum burden, as experimental depletion of these cells over this timeframe now led to productive infections. Taken together, this study identifies that murine models of menstruation can be a valuable tool for investigating how the menstrual cycle modulates immune homeostasis and for identifying ways to strengthen mucosal immune defenses against genitourinary pathogens in women.

immunology↗

Detuning of the Ribosome Conformational Landscape Promotes Antibiotic Resistance and Collateral Sensitivity

Around 50% of the current antibiotic arsenal targets the ribosome, thus resistance to ribosome-targeting antibiotics poses severe challenges to antimicrobial treatments. Here, we characterize a 12-nucleotide deletion in the rplF gene encoding the uL6 ribosomal protein, which was identified in a tobramycin-resistant strain of Pseudomonas aeruginosa isolated from a cystic fibrosis patient. To understand this resistance, we determined 87 cryo-EM structures of wild-type and mutant ribosomes characterizing their conformational landscape. Our analysis reveals how detuning of the ribosome dynamics alters its rotational movement circumventing tobramycin inhibition. The mutation compromises the 50S assembly, triggering structural instability and inducing a different rotational dynamic of the 70S. We found 4 new binding sites of tobramycin, one of them exclusive of the mutant, where the binding of the antibiotic acts as an allosteric activator skipping inhibition. Our data also illustrate how chloramphenicol stabilizes the mutant ribosome, enhancing inhibition and thereby leading to collateral sensitivity.

biophysics↗

Transcriptomic Evaluation of Stress Vulnerability Network using Single Cell RNA-Seq in mouse Prefrontal Cortex

Increased vulnerability to stress is a major risk factor for the manifestation of several mood disorders, including major depressive disorder (MDD). Despite the status of MDD as a significant donor to global disability, the complex integration of genetic and environmental factors that contribute to the behavioral display of such disorders has made a thorough understanding of related etiology elusive. Recent developments suggest that a brain-wide network approach is needed, taking into account the complex interplay of cell types spanning multiple brain regions. Single cell RNA-sequencing technologies can provide transcriptomic profiling at the single-cell level across heterogenous samples. Furthermore, we have previously used local field potential oscillations and machine learning to identify an electrical brain network that is indicative of a predisposed vulnerability state. Thus, this study combined single cell RNA-sequencing (scRNA-Seq) with electrical brain network measures of the stress-vulnerable state, providing a unique opportunity to access the relationship between stress network activity and transcriptomic changes within individual cell types. We found especially high numbers of differentially expressed genes between animals with high and low stress vulnerability brain network activity in astrocytes and glutamatergic neurons but we estimated that vulnerability network activity depends most on GABAergic neurons. High vulnerability network activity included upregulation of microglia and mitochondrial and metabolic pathways, while lower vulnerability involved synaptic regulation. Genes that were differentially regulated with vulnerability network activity significantly overlapped with genes identified as having significant SNPs by human GWAS for depression. Taken together, these data provide the gene expression architecture of a previously uncharacterized stress vulnerability brain state, enabling new understanding and intervention of predisposition to stress susceptibility. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/540705v2_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@1ebe036org.highwire.dtl.DTLVardef@cd2131org.highwire.dtl.DTLVardef@13e3519org.highwire.dtl.DTLVardef@1100e62_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

A multiadjuvant polysaccharide-amino acid-lipid (PAL) subunit nanovaccine generates robust systemic and lung-specific mucosal immune responses against SARS-CoV-2 in mice

Existing parenteral SARS-CoV-2 vaccines produce only limited mucosal responses, which are essential for reducing transmission and achieving sterilizing immunity. Appropriately designed mucosal boosters could overcome the shortcomings of parenteral vaccines and enhance pre- existing systemic immunity. Here we present a new protein subunit nanovaccine using multiadjuvanted (e.g. RIG-I: PUUC, TLR9: CpG) polysaccharide-amino acid-lipid nanoparticles (PAL-NPs) that can be delivered both intramuscularly (IM) and intranasally (IN) to generate balanced mucosal-systemic SARS-CoV-2 immunity. Mice receiving IM-Prime PUUC+CpG PAL- NPs, followed by an IN-Boost, developed high levels of IgA, IgG, and cellular immunity in the lung, and showed robust systemic humoral immunity. Interestingly, as a purely intranasal vaccine (IN-Prime/IN-Boost), PUUC+CpG PAL-NPs induced stronger lung-specific T cell immunity than IM-Prime/IN-Boost, and a comparable IgA and neutralizing antibodies, although with a lower systemic antibody response, indicating that a fully mucosal delivery route for SARS-CoV-2 vaccination may also be feasible. Our data suggest that PUUC+CpG PAL-NP subunit vaccine is a promising candidate for generating SARS-CoV-2 specific mucosal immunity.

bioengineering↗

The structure of anticorrelated networks in the human brain

During the performance of a specific task or at rest, the activity of different brain regions shares statistical dependencies that reflect functional connections. While these relationships have been studied intensely for positively correlated networks, considerably less attention has been paid to negatively correlated networks, a.k.a. anticorrelated networks (ACNs). Here, we have addressed this issue by making use of two neuroimaging datasets: one of N=192 young healthy adults; and another of N=40 subjects that was divided into two groups of young and old participants. We first provided a full description of the anatomical composition of the different ACNs, each of which participated in distinct resting-state networks (RSNs). In terms of their frequency of participation, from highest to the lowest, the major anticorrelated brain areas are the precuneus, the anterior supramarginal gyrus and the central opercular cortex. Subsequently, by evaluating the more detailed structure of ACNs, we show it is possible to find significant differences in these in association with specific conditions, in particular by comparing groups of young and old participants. Our main finding is that of increased anticorrelation for cerebellar interactions in older subjects. Overall, our results give special relevance to ACNs and they suggest they may serve to disentangle unknown alterations in certain conditions, as might occur in neurodegenerative diseases at early onset or in some psychiatric conditions.

neuroscience↗

High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

In recent years, the development of new image analysis approaches has highlighted the possibility of recovering superresolution information from short sequences of wide-field images. Our recently developed method, SRRF (Super-Resolution Radial Fluctuations), enables long-term live-cell imaging beyond the resolution limit without specialized hardware. Here, we present eSRRF (enhanced-SRRF), a significant improvement over our initial method, enhancing image fidelity to the underlying structure and resolution. Especially, eSRRF uses automated data-driven parameter optimization, including an estimation of the number of frames necessary for optimal reconstruction. We demonstrate the improved fidelity of the images reconstructed with eSRRF and highlight its versatility and ease of use over a wide range of microscopy techniques and biological systems. We also extend eSRRF to 3D super-resolution microscopy by combining it with multi-focus microscopy (MFM), obtaining volumetric super-resolution imaging of live cells with acquisition speed of ~1 volume/second.

biophysics↗

Senescent stroma induces nuclear deformations in cancer cells via the inhibition of RhoA/ROCK/myosin II-based cytoskeletal tension

The presence of senescent cells within tissues has been functionally linked to malignant transformations. Here, using tension-gauge tethers technology, particle-tracking microrheology, and quantitative microscopy, we demonstrate that senescent associated secretory phenotype (SASP) derived from senescent fibroblasts impose nuclear lobulations and volume shrinkage on malignant cells, which stems from the loss of RhoA/ROCK/myosin II-based cortical tension. This loss in cytoskeletal tension induces decreased cellular contractility, adhesion, and increased mechanical compliance. These SASP-induced morphological changes are in part mediated by lamin A/C. These findings suggest that SASP induces a defective outside-in mechanotransduction, from actomyosin fibers in the cytoplasm to the nuclear lamina, thereby triggering a cascade of biophysical and biomolecular changes in cells that associate with malignant transformations.

cancer biology↗

Antibody Conversion rates to SARS-CoV-2 in Saliva from Children Attending Summer Schools in Barcelona, Spain

Surveillance tools to estimate infection rates in young populations are essential to guide recommendations for school reopening and management during viral epidemics. Ideally, field-deployable non-invasive, sensitive techniques are required to detect low viral load exposures among asymptomatic children. We determined SARS-CoV-2 antibody conversion by high-throughput Luminex assays in saliva samples collected weekly in 1,509 children and 396 adults in 22 Summer schools and 2 pre-schools in 27 venues in Barcelona, Spain, from June 29th to July 31st 2020, between the first and second COVID-19 pandemic waves. Saliva antibody conversion defined as [≥]4-fold increase in IgM, IgA and/or IgG levels to SARS-CoV-2 antigens between two visits over a 5-week period was 3.22% (49/1518), or 2.36% if accounting for potentially cross-reactive antibodies, six times higher than the cumulative infection rate (0.53%) by weekly saliva RT-PCR screening. IgG conversion was higher in adults (2.94%, 11/374) than children (1.31%, 15/1144) (p=0.035), IgG and IgA levels moderately increased with age, and antibodies were higher in females. Most antibody converters increased both IgG and IgA antibodies but some augmented either IgG or IgA, with a faster decay over time for IgA than IgG. Nucleocapsid rather than spike was the main antigen target. Anti-spike antibodies were significantly higher in individuals not reporting symptoms than symptomatic individuals, suggesting a protective role against COVID-19. To conclude, saliva antibody profiling including three isotypes and multiplexing antigens is a useful and more user-friendly tool for screening pediatric populations to determine SARS-CoV-2 exposure and guide public health policies during pandemics.

immunology↗