Clonal structure and the specificity of vaccine-induced T cell response to SARS-CoV-2 Spike protein.

Adenovirus vaccines, particularly the COVID-19 Ad5-nCoV adenovirus vaccine, have emerged as promising tools in the fight against infectious diseases. In this study, we investigated the structure of the T cell response to the Spike protein of the SARS-CoV-2 virus used in the COVID-19 Ad5-nCoV adenoviral vaccine in a phase 3 clinical trial (NCT04540419). In 69 participants, we collected peripheral blood samples at four time points after vaccination or placebo injection. Sequencing of T cell receptor repertoires from Spike-stimulated T cell cultures at day 14 from 17 vaccinated revealed a more diverse CD4+ T cell repertoire compared to CD8+. Nevertheless, CD8+ clonotypes accounted for more than half of the Spike-specific repertoire. Our longitudinal analysis showed a peak T cell response at day 14, followed by a decline until month 6. Remarkably, multiple T cell clonotypes persisted for at least 6 months after vaccination, as demonstrated by ex vivo stimulation. Examination of CDR3 regions revealed homologous sequences in both CD4+ and CD8+ clonotypes, with major CD8+ clonotypes sharing high similarity with annotated sequences specific for the NYNYLYRLF peptide, suggesting potential immunodominance. In conclusion, our study demonstrates the immunogenicity of the Ad5-nCoV adenoviral vaccine and highlights its ability to induce robust and durable T cell responses. These findings provide valuable insight into the efficacy of the vaccine against COVID-19 and provide critical information for ongoing efforts to control infectious diseases.

Clonal diversity predicts persistence of SARS-CoV-2 epitope-specific T-cell response.

T cells play a pivotal role in reducing disease severity during SARS-CoV-2 infection and formation of long-term immune memory. We studied 50 COVID-19 convalescent patients and found that T cell response was induced more frequently and persisted longer than circulating antibodies. We identified 756 clonotypes specific to nine CD8+ T cell epitopes. Some epitopes were recognized by highly similar public clonotypes. Receptors for other epitopes were extremely diverse, suggesting alternative modes of recognition. We tracked persistence of epitope-specific response and individual clonotypes for a median of eight months after infection. The number of recognized epitopes per patient and quantity of epitope-specific clonotypes decreased over time, but the studied epitopes were characterized by uneven decline in the number of specific T cells. Epitopes with more clonally diverse TCR repertoires induced more pronounced and durable responses. In contrast, the abundance of specific clonotypes in peripheral circulation had no influence on their persistence.

Look and ye shall hear: Selective auditory attention modulates the audiovisual correspondence effect.

One of the unresolved questions in multisensory research is that of automaticity of consistent associations between sensory features from different modalities (e.g. high visual locations associated with high sound pitch). We addressed this issue by examining a possible role of selective attention in the audiovisual correspondence effect. We orthogonally manipulated loudness and pitch, directing participants' attention to the auditory modality only and using pitch and loudness identification tasks. Visual stimuli in high, low or central spatial locations appeared simultaneously with the sounds. If the correspondence effect is automatic, it should not be affected by task changes. The results, however, demonstrated a cross-modal pitch-verticality correspondence effect only when participants' attention was directed to pitch, but not to loudness identification task; moreover, the effect was present only in the upper location. The findings underscore the involvement of selective attention in cross-modal associations and support a top-down account of audiovisual correspondence effects.

Explicit encoding vs. fast mapping of novel spoken words: Electrophysiological and behavioural evidence of diverging mechanisms.

It has been claimed that two major neurocognitive mechanisms - instruction-based explicit encoding (EE) and inference-driven fast mapping (FM) may be involved in rapid acquisition of novel words, but their exact neural underpinnings remain poorly understood. To address this, we trained 36 adult participants with 20 novel spoken words in an audio-visual task, carefully balanced between the EE and FM conditions for physical, psycholinguistic and pragmatic properties as well as the overall task setup. To assess the neural dynamics associated with novel word acquisition, we recorded event-related potentials (ERPs) elicited by these words before and after training, and analysed their relationship with the behavioural learning outcomes, measured in a semantic matching task. Both learning regimes led to successful acquisition, which was somewhat more efficient for EE than FM, as indicated by higher accuracy in the behavioural task. We also found that, whereas words learnt via both EE and FM protocols elicited most pronounced ERP peaks at ∼196 and ∼280 ms, these two phases of activity diverged with respect to the learning type. Multiple linear regression and correlation analyses indicated that the learning-induced amplitude dynamics in the earlier peak was significantly related to behavioural performance for FM-learned items, which may possibly be explained by FM's stronger reliance on early automatic mechanisms of word processing. Performance on EE words was, in turn, significantly linked to the amplitude of the second peak only, potentially due to the involvement of later, top-down controlled processes in this type of word acquisition. Grand-average ERP-based source analysis indicated a left-lateralised activity in the anterior-temporal lobe for FM learning, and a bilateral activation for EE. The results confirm the existence of partially diverging neurocognitive systems for word acquisition and suggest that the configuration of newly established word memory circuits depends on the mode of their acquisition.

Immunogenic epitope panel for accurate detection of non-cross-reactive T cell response to SARS-CoV-2.

The ongoing COVID-19 pandemic calls for more effective diagnostic tools. T cell response assessment serves as an independent indicator of prior COVID-19 exposure while also contributing to a more comprehensive characterization of SARS-CoV-2 immunity. In this study, we systematically assessed the immunogenicity of 118 epitopes with immune cells collected from multiple cohorts of vaccinated, convalescent, healthy unexposed, and SARS-CoV-2-exposed donors. We identified 75 immunogenic epitopes, 24 of which were immunodominant. We further confirmed HLA restriction for 49 epitopes and described association with more than 1 HLA allele for 14 of these. Exclusion of 2 cross-reactive epitopes that generated a response in prepandemic samples left us with a 73-epitope set that offered excellent diagnostic specificity without losing sensitivity compared with full-length antigens, and this evoked a robust cross-reactive response. We subsequently incorporated this set of epitopes into an in vitro diagnostic Corona-T-test, which achieved a diagnostic accuracy of 95% in a clinical trial. In a cohort of asymptomatic seronegative individuals with a history of prolonged SARS-CoV-2 exposure, we observed a complete absence of T cell response to our epitope panel. In combination with strong reactivity to full-length antigens, this suggests that a cross-reactive response might protect these individuals.

Anodal tDCS over Broca's area improves fast mapping and explicit encoding of novel vocabulary.

An accumulating body of evidence suggests that transcranial direct current stimulation (tDCS) can be used to affect language processing, including word acquisition. There has been, however, no comprehensive study of effects of tDCS of the core language areas in relation to the main word-learning mechanisms. Two principal strategies have been posited as important for natural word acquisition: explicit encoding (EE) which relies on direct instructions and repetition of material, and fast mapping (FM) which operates implicitly, via context-based inference or deduction. We used anodal and cathodal tDCS of Broca's and Wernicke's areas to assess effects of stimulation site and polarity on novel word acquisition in both EE and FM regimes. 160 participants, divided into five groups, received 15 min of cathodal or anodal tDCS over one of the two areas or a sham (placebo) stimulation before learning eight novel words, presented ten times each in a short naturalistic audio-visual word-picture association session, fully counterbalanced across different learning regimes. The outcome of novel word acquisition was measured immediately after the training using a free recall task. The results showed elevated accuracy in all real stimulation groups in comparison with sham stimulation; however, this effect only reached full significance after anodal tDCS of Broca's area. Comparisons between the two learning modes indicated that Broca's anodal tDCS significantly improved both implicit and explicit acquisition of novel vocabulary in comparison with sham tDCS, without, however, any significant differences between EE and FM regimes as such. The results indicate involvement of the left inferior-frontal neocortex in the learning of novel vocabulary and suggest a possibility to promote different types of word acquisition using anodal tDCS of this area.

Corrigendum: Just Open Your Mind? A Randomized, Controlled Study on the Effects of Meditation on Creativity.

[This corrects the article DOI: 10.3389/fpsyg.2021.663881.].

Just Open Your Mind? A Randomized, Controlled Study on the Effects of Meditation on Creativity.

Creativity is a crucial prerequisite for innovation, successful problem solving, and self-expression, but how do we affect creative thinking in a positive way? The present study investigated the effects of open monitoring meditation (OMM) on creativity. We proposed that OMM will benefit creativity in metaphor production by cognitive flexibility (CF) enhancement. In the main study, participants were randomly assigned to one of the three groups: meditation, active, and passive controls. The first two groups performed an audio-guided task (real meditation or a narrative on house plants) for 2 weeks, and the third one had no task. Pre- and post-tests included measures of metaphor production, CF, state, sustained attention, attention shifting, and intelligence. We found no significant intra- or intergroup differences that would suggest OMM effects on creativity. Further, no links were found between measures of metaphor creativity and CF. Findings reveal potential challenges of using meditation as a cognitive enhancement tool. Methodological issues concerning meditation research, as well as creativity and CF measures, are discussed.

Electrophysiological Evidence of Dissociation Between Explicit Encoding and Fast Mapping of Novel Spoken Words.

Existing behavioral, neuropsychological and functional neuroimaging data suggest that at least two major cognitive strategies are used for new word learning: fast mapping (FM) via context-dependent inference and explicit encoding (EE) via direct instruction. However, these distinctions remain debated at both behavioral and neurophysiological levels, not least due to confounds related to diverging experimental settings. Furthermore, the neural dynamics underpinning these two putative processes remain poorly understood. To tackle this, we designed a paradigm presenting 20 new spoken words in association with pictures in either FM or EE settings, closely matched for auditory and visual features and overall task demands. We tested word acquisition using a range of behavioral measures as well as passive event-related potential (ERP) responses, an established measure of word memory trace activation, and compared brain activity elicited by novel FM and EE words before and after the learning session. Behavioral data obtained in free recall, recognition and semantic word-picture matching tasks indicated successful acquisition of new words after just 10 exposures. Crucially, we found no behavioral evidence of different acquisition outcomes between FM and EE learning. ERP data, which exhibited the main response peaks at ~170, 250, and 520 ms, also indicated successful learning, with statistically different responses between novel and familiar words present only before, but not after the training, suggesting rapid formation of new neural memory circuits matching in activation those for previously known words. Furthermore, already at the earliest peak, we found different topographic distributions for the two learning types, with left-lateralized FM dynamics, suggestive of core language system involvement, and more diffuse activity for EE items, possibly suggesting the role of attention/executive control network. A similar effect also manifested later, at ~520 ms. Our data suggest that while both EE and FM learning can be successful for rapid word acquisition at the behavioral level, the diverging electrophysiological patterns suggest a dissociation between the neural systems underpinning these learning strategies.

Psychometric Comparisons of Benevolent and Corrective Humor across 22 Countries: The Virtue Gap in Humor Goes International.

Recently, two forms of virtue-related humor, benevolent and corrective, have been introduced. Benevolent humor treats human weaknesses and wrongdoings benevolently, while corrective humor aims at correcting and bettering them. Twelve marker items for benevolent and corrective humor (the BenCor) were developed, and it was demonstrated that they fill the gap between humor as temperament and virtue. The present study investigates responses to the BenCor from 25 samples in 22 countries (overall N = 7,226). The psychometric properties of the BenCor were found to be sufficient in most of the samples, including internal consistency, unidimensionality, and factorial validity. Importantly, benevolent and corrective humor were clearly established as two positively related, yet distinct dimensions of virtue-related humor. Metric measurement invariance was supported across the 25 samples, and scalar invariance was supported across six age groups (from 18 to 50+ years) and across gender. Comparisons of samples within and between four countries (Malaysia, Switzerland, Turkey, and the UK) showed that the item profiles were more similar within than between countries, though some evidence for regional differences was also found. This study thus supported, for the first time, the suitability of the 12 marker items of benevolent and corrective humor in different countries, enabling a cumulative cross-cultural research and eventually applications of humor aiming at the good.

The CDC42-interacting protein 4 controls epithelial cell cohesion and tumor dissemination.

The role of endocytic proteins and the molecular mechanisms underlying epithelial cell cohesion and tumor dissemination are not well understood. Here, we report that the endocytic F-BAR-containing CDC42-interacting protein 4 (CIP4) is required for ERBB2- and TGF-ß1-induced cell scattering, breast cancer (BC) cell motility and invasion into 3D matrices, and conversion from ductal breast carcinoma in situ to invasive carcinoma in mouse xenograft models. CIP4 promotes the formation of an E-cadherin-CIP4-SRC complex that controls SRC activation, E-cadherin endocytosis, and localized phosphorylation of the myosin light chain kinase, thereby impinging on the actomyosin contractility required to generate tangential forces to break cell-cell junctions. CIP4 is upregulated in ERBB2-positive human BC, correlates with increased distant metastasis, and is an independent predictor of poor disease outcome in subsets of BC patients. Thus, it critically controls cell-cell cohesion and is required for the acquisition of an invasive phenotype in breast tumors.

Large, controllable spikes of magnetoresistance in La(2/3)Ca(1/3)MnO3/SrTiO3 superlattices.

We have investigated superlattices consisting of up to 30 epitaxial nanomultilayers (3-7 nm thick) of ferromagnetic La(2/3)Ca(1/3)MnO(3) (LCMO) and insulating SrTiO(3) (STO) hybrids. The superlattices demonstrate dramatic shifts of Curie temperature, indicating the possibility of its tunability. The metal-insulator transition (MIT) has been observed around 140 K. Below the MIT temperature, the superlattices have shown sharp drops of resistivity, facilitating the largest and sharpest magnetoresistance peaks (>2000%) ever observed in LCMO films and superlattices at low temperatures. The observed experimental results can be explained in the frame of the phase separation model in manganites with well-organized structures. The results of magnetic and transport measurements of such hybrid structures are discussed, indicating a magnetodielectric effect in STO interlayers. The magnetic and transport properties of the superlattices are shown to be technology-dependent, experiencing dimensional transitions, which enables the creation of structures with prescribed magnetoresistance characteristics for a broad range of applications.

E-cadherin is under constitutive actomyosin-generated tension that is increased at cell-cell contacts upon externally applied stretch.

Classical cadherins are transmembrane proteins at the core of intercellular adhesion complexes in cohesive metazoan tissues. The extracellular domain of classical cadherins forms intercellular bonds with cadherins on neighboring cells, whereas the cytoplasmic domain recruits catenins, which in turn associate with additional cytoskeleton binding and regulatory proteins. Cadherin/catenin complexes are hypothesized to play a role in the transduction of mechanical forces that shape cells and tissues during development, regeneration, and disease. Whether mechanical forces are transduced directly through cadherins is unknown. To address this question, we used a Förster resonance energy transfer (FRET)-based molecular tension sensor to test the origin and magnitude of tensile forces transmitted through the cytoplasmic domain of E-cadherin in epithelial cells. We show that the actomyosin cytoskeleton exerts pN-tensile force on E-cadherin, and that this tension requires the catenin-binding domain of E-cadherin and αE-catenin. Surprisingly, the actomyosin cytoskeleton constitutively exerts tension on E-cadherin at the plasma membrane regardless of whether or not E-cadherin is recruited to cell-cell contacts, although tension is further increased at cell-cell contacts when adhering cells are stretched. Our findings thus point to a constitutive role of E-cadherin in transducing mechanical forces between the actomyosin cytoskeleton and the plasma membrane, not only at cell-cell junctions but throughout the cell surface.

Kinase-related protein/telokin inhibits Ca2+-independent contraction in Triton-skinned guinea pig taenia coli.

KRP (kinase-related protein), also known as telokin, has been proposed to inhibit smooth muscle contractility by inhibiting the phosphorylation of the rMLC (regulatory myosin light chain) by the Ca2+-activated MLCK (myosin light chain kinase). Using the phosphatase inhibitor microcystin, we show in the present study that KRP also inhibits Ca2+-independent rMLC phosphorylation and smooth muscle contraction mediated by novel Ca2+-independent rMLC kinases. Incubating KRP-depleted Triton-skinned taenia coli with microcystin at pCa>8 induced a slow contraction reaching 90% of maximal force (Fmax) at pCa 4.5 after approximately 25 min. Loading the fibres with KRP significantly slowed down the force development, i.e. the time to reach 50% of Fmax was increased from 8 min to 35 min. KRP similarly inhibited rMLC phosphorylation of HMM (heavy meromyosin) in vitro by MLCK or by the constitutively active MLCK fragment (61K-MLCK) lacking the myosin-docking KRP domain. A C-terminally truncated KRP defective in myosin binding inhibited neither force nor HMM phosphorylation. Phosphorylated KRP inhibited the rMLC phosphorylation of HMM in vitro and Ca2+-insensitive contractions in fibres similar to unphosphorylated KRP, whereby the phosphorylation state of KRP was not altered in the fibres. We conclude that (i) KRP inhibits not only MLCK-induced contractions, but also those elicited by Ca2+-independent rMLC kinases; (ii) phosphorylation of KRP does not modulate this effect; (iii) binding of KRP to myosin is essential for this inhibition; and (iv) KRP inhibition of rMLC phosphorylation is most probably due to the shielding of the phosphorylation site on the rMLC.

Regulation of G-protein coupled receptor traffic by an evolutionary conserved hydrophobic signal.

Plasma membrane (PM) expression of G-protein coupled receptors (GPCRs) is required for activation by extracellular ligands; however, mechanisms that regulate PM expression of GPCRs are poorly understood. For some GPCRs, such as alpha2c-adrenergic receptors (alpha(2c)-ARs), heterologous expression in non-native cells results in limited PM expression and extensive endoplasmic reticulum (ER) retention. Recently, ER export/retentions signals have been proposed to regulate cellular trafficking of several GPCRs. By utilizing a chimeric alpha(2a)/alpha(2c)-AR strategy, we identified an evolutionary conserved hydrophobic sequence (ALAAALAAAAA) in the extracellular amino terminal region that is responsible in part for alpha(2c)-AR subtype-specific trafficking. To our knowledge, this is the first luminal ER retention signal reported for a GPCR. Removal or disruption of the ER retention signal dramatically increased PM expression and decreased ER retention. Conversely, transplantation of this hydrophobic sequence into alpha(2a)-ARs reduced their PM expression and increased ER retention. This evolutionary conserved hydrophobic trafficking signal within alpha(2c)-ARs serves as a regulator of GPCR trafficking.

Organization of beta-adrenoceptor signaling compartments by sympathetic innervation of cardiac myocytes.

The sympathetic nervous system regulates cardiac function through the activation of adrenergic receptors (ARs). beta(1) and beta(2)ARs are the primary sympathetic receptors in the heart and play different roles in regulating cardiac contractile function and remodeling in response to injury. In this study, we examine the targeting and trafficking of beta(1) and beta(2)ARs at cardiac sympathetic synapses in vitro. Sympathetic neurons form functional synapses with neonatal cardiac myocytes in culture. The myocyte membrane develops into specialized zones that surround contacting axons and contain accumulations of the scaffold proteins SAP97 and AKAP79/150 but are deficient in caveolin-3. The beta(1)ARs are enriched within these zones, whereas beta(2)ARs are excluded from them after stimulation of neuronal activity. The results indicate that specialized signaling domains are organized in cardiac myocytes at sites of contact with sympathetic neurons and that these domains are likely to play a role in the subtype-specific regulation of cardiac function by beta(1) and beta(2)ARs in vivo.

Differential targeting and function of alpha2A and alpha2C adrenergic receptor subtypes in cultured sympathetic neurons.

Previous research suggested that alpha2A and alpha2C adrenergic receptor (AR) subtypes have overlapping but unique physiological roles in neuronal signaling; however, the basis for these dissimilarities is not completely known. To better understand the observed functional differences between these autoreceptors, we investigated targeting and signaling of endogenously expressed alpha2A and alpha2CARs in cultured sympathetic ganglion neurons (SGN). At Days 1 and 4, alpha2A and alpha2CARs could be readily detected in SGN from wild-type mice. By Day 8, alpha2A ARs were targeted to cell body, as well as axonal and dendritic sites, whereas alpha2C ARs were primarily localized to an intracellular vesicular pool within the cell body and proximal dendritic projections. Expression of synaptic vesicle marker protein SV2 did not differ at Day 8 nor co-localize with either subtype. By Day 16, however, alpha2C ARs had relocated to somatodendritic and axonal sites and, unlike alpha2A ARs, co-localized with SV2 at synaptic contact sites. Consistent with a functional role for alpha2 ARs, we also observed that dexmedetomidine stimulation of cultured SGN more efficiently inhibited depolarization-induced calcium entry into older, compared to younger, cultures. These results provide direct evidence of distinct developmental patterns of endogenous alpha2A and alpha2C AR targeting and function in a native cell system and that maturation of SGN in culture leads to alterations in neuronal properties required for proper targeting. More importantly, the co-localization at Day 16 of alpha2C ARs at sites of synaptic contact may partially explain the differential modulation of neurotransmitter release and responsiveness to action potential frequency observed between alpha2A and alpha2C ARs in SGN.