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The human Betacoronavirus OC43 is a common cause of respiratory viral infections in adults and children. Lung infections with OC43 are associated with mortality, especially in hematopoietic stem cell transplant recipients. Neutralizing antibodies play a major role in protection against many respiratory viral infections, but to date a live viral neutralization assay for OC43 has not been described. We isolated a human monoclonal antibody (OC2) that binds to the spike protein of OC43 and neutralizes the live virus derived from the original isolate of OC43. We used this monoclonal antibody to develop and test the performance of two readily accessible in vitro assays for measuring antibody neutralization, one utilizing cytopathic effect and another utilizing an ELISA of infected cells. We used both methods to measure the neutralizing activity of the OC2 monoclonal antibody and of human plasma. These assays could prove useful for studying humoral responses to OC43 and cross-neutralization with other medically important betacoronaviruses.

Forecasting emergent pest spread is paramount to mitigating their impacts. For host-specialized pests, epidemiological models of spread through a single host population are well developed. However, most pests attack multiple host species; the challenge is predicting which communities are most vulnerable to infestation. Here, we develop a phylogenetically-informed approach to predict establishment of emergent multi-host pests across heterogeneous landscapes. We model a beetle-pathogen symbiotic complex on trees, introduced from Southeast Asia to California. The phyloEpi model for likelihood of establishment was predicted from the phylogenetic composition of woody species in the invaded community and the influence of temperature on beetle reproduction. Plant communities dominated by close relatives of known epidemiologically critical hosts were four times more likely to become infested than communities with more distantly related species. Where microclimate favored beetle reproduction, pest establishment was greater than expected based only on species composition. We applied this phyloEpi model to predict infestation risk in California using weather data and complete tree inventories from 9262 1-km 2 grids in 170 cities. Regions in the state predicted with low likelihood of infestation were confirmed by independent monitoring. Analysts can adapt these phylogenetic ecology tools to predict spread of any multi-host pest in novel habitats. A study demonstrates how a phylogenetic epidemiology approach can be used to improve rapid responses against emergent pests, which are mostly able to attack multiple host species and spread through heterogeneous landscapes.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) hybrid immunity is more protective than vaccination or previous infection alone. To investigate the kinetics of spike-reactive T (T S ) cells from SARS-CoV-2 infection through messenger RNA vaccination in persons with hybrid immunity, we identified the T cell receptor (TCR) sequences of thousands of index T S cells and tracked their frequency in bulk TCRβ repertoires sampled longitudinally from the peripheral blood of persons who had recovered from coronavirus disease 2019 (COVID-19). Vaccinations led to large expansions in memory T S cell clonotypes, most of which were CD8 + T cells, while also eliciting diverse T S cell clonotypes not observed before vaccination. TCR sequence similarity clustering identified public CD8 + and CD4 + TCR motifs associated with spike (S) specificity. Synthesis of longitudinal bulk ex vivo single-chain TCRβ repertoires and paired-chain TCRɑβ sequences from droplet sequencing of T S cells provides a roadmap for the rapid assessment of T cell responses to vaccines and emerging pathogens. Koelle and colleagues use an activation marker-dependent approach to determine the recruitment of TCR by three doses of mRNA vaccination in individuals previously infected with SARS-CoV-2.

The ectocervix is part of the lower female reproductive tract (FRT), which is susceptible to sexually transmitted infections (STIs). Comprehensive knowledge of the phenotypes and T cell receptor (TCR) repertoire of tissue-resident memory T cells (TRMs) in the human FRT is lacking. We took single-cell RNA-Seq approaches to simultaneously define gene expression and TCR clonotypes of the human ectocervix. There were significantly more CD8+ than CD4+ T cells. Unsupervised clustering and trajectory analysis identified distinct populations of CD8+ T cells with IFNGhiGZMBloCD69hiCD103lo or IFNGloGZMBhiCD69medCD103hi phenotypes. Little overlap was seen between their TCR repertoires. Immunofluorescence staining showed that CD103+CD8+ TRMs were preferentially localized in the epithelium, whereas CD69+CD8+ TRMs were distributed evenly in the epithelium and stroma. Ex vivo assays indicated that up to 14% of cervical CD8+ TRM clonotypes were HSV-2 reactive in HSV-2-seropositive persons, reflecting physiologically relevant localization. Our studies identified subgroups of CD8+ TRMs in the human ectocervix that exhibited distinct expression of antiviral defense and tissue residency markers, anatomic locations, and TCR repertoires that target anatomically relevant viral antigens. Optimization of the location, number, and function of FRT TRMs is an important approach for improving host defenses to STIs.