Recent data suggest that facultative hypothermic responses such as torpor are more important in the energy balance of birds from tropical and sub‐tropical regions than previously thought. We used telemetric measurements of skin temperature (Tskin) for five individuals on 151 bird‐nights to investigate the occurrence of torpor during winter in an 81 g African caprimulgid, the freckled nightjar Caprimulgus tristigma. We found that freckled nightjars have the capacity to enter torpor, with a minimum observed Tskin of 12.8°C. During the torpor bouts we observed, complete rewarming typically occurred after sunrise, and coincided with the availability of solar radiation. There was considerable inter‐individual variability in the frequency and depth of torpor bouts, with one female nightjar exhibiting particularly frequent and deep torpor. Our results confirm the ability to use torpor by a nocturnal aerial insectivore from the Afrotropics, and reiterate the variability in patterns of torpor that can exist within a population.

Background In Sweden, social restrictions to contain SARS‐CoV‐2 have primarily relied upon voluntary adherence to a set of recommendations. Strict lockdowns have not been enforced, potentially affecting viral dissemination. To understand the levels of past SARS‐CoV‐2 infection in the Stockholm population before the start of mass vaccinations, healthy blood donors and pregnant women ( n = 5,100) were sampled at random between 14 March 2020 and 28 February 2021. Methods In this cross‐sectional prospective study, otherwise‐healthy blood donors ( n = 2,600) and pregnant women ( n = 2,500) were sampled for consecutive weeks (at four intervals) throughout the study period. Sera from all participants and a cohort of historical (negative) controls ( n = 595) were screened for IgG responses against stabilized trimers of the SARS‐CoV‐2 spike (S) glycoprotein and the smaller receptor‐binding domain (RBD). As a complement to standard analytical approaches, a probabilistic (cut‐off independent) Bayesian framework that assigns likelihood of past infection was used to analyse data over time. Setting Healthy participant samples were randomly selected from their respective pools through Karolinska University Hospital. The study was carried out in accordance with Swedish Ethical Review Authority: registration number 2020–01807. Participants No participants were symptomatic at sampling, and blood donors were all over the age of 18. No additional metadata were available from the participants. Results Blood donors and pregnant women showed a similar seroprevalence. After a steep rise at the start of the pandemic, the seroprevalence trajectory increased steadily in approach to the winter second wave of infections, approaching 15% of all individuals surveyed by 13 December 2020. By the end of February 2021, 19% of the population tested seropositive. Notably, 96% of seropositive healthy donors screened ( n = 56) developed neutralizing antibody responses at titres comparable to or higher than those observed in clinical trials of SARS‐CoV‐2 spike mRNA vaccination, supporting that mild infection engenders a competent B‐cell response. Conclusions These data indicate that in the first year since the start of community transmission, seropositivity levels in metropolitan in Stockholm had reached approximately one in five persons, providing important baseline seroprevalence information prior to the start of vaccination.

The COVID-19 pandemic has posed a tremendous challenge for the global community. We established a translational approach combining home blood sampling by finger-pricking with multiplexed serology to assess the exposure to the SARS-CoV-2 virus in a general population. The developed procedure determines the immune response in multiplexed assays against several spike (S, here denoted SPK), receptor binding domain (RBD) and nucleocapsid (NCP) proteins in eluates from dried capillary blood. The seroprevalence was then determined in two study sets by mailing 1000 blood sampling kits to random households in urban Stockholm during early and late April 2020, respectively. After receiving 55% (1097/2000) of the cards back within three weeks, 80% (878/1097) were suitable for the analyses of IgG and IgM titers. The data revealed diverse pattern of immune response, thus seroprevalence was dependent on the antigen, immunoglobulin class, stringency to include different antigens, as well as the required analytical performance. Applying unsupervised dimensionality reduction to the combined IgG and IgM data, 4.4% (19/435; 95% CI: 2.4%-6.3%) and 6.3% (28/443; 95% CI: 4.1%-8.6%) of the samples clustered with convalescent controls. Using overlapping scores from at least two SPK antigens, prevalence rates reached 10.1% (44/435; 95% CI: 7.3%-12.9%) in study set 1 and 10.8% (48/443; 95% CI: 7.9%-13.7%). Measuring the immune response against several SARS-CoV-2 proteins in a multiplexed workflow can provide valuable insights about the serological diversity and improve the certainty of the classification. Combining such assays with home-sampling of blood presents a viable strategy for individual-level diagnostics and towards an unbiased assessment of the seroprevalence in a population and may serve to improve our understanding about the diversity of COVID-19 etiology.

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Serological testing is essential to curb the consequences of the COVID-19 pandemic. However, most assays are still limited to single analytes and samples collected within healthcare. Thus, we establish a multianalyte and multiplexed approach to reliably profile IgG and IgM levels against several versions of SARS-CoV-2 proteins (S, RBD, N) in home-sampled dried blood spots (DBS). We analyse DBS collected during spring of 2020 from 878 random and undiagnosed individuals from the population in Stockholm, Sweden, and use classification approaches to estimate an accumulated seroprevalence of 12.5% (95% CI: 10.3%–14.7%). This includes 5.4% of the samples being IgG+IgM+ against several SARS-CoV-2 proteins, as well as 2.1% being IgG−IgM+ and 5.0% being IgG+IgM− for the virus’ S protein. Subjects classified as IgG+ for several SARS-CoV-2 proteins report influenza-like symptoms more frequently than those being IgG+ for only the S protein (OR = 6.1; p < 0.001). Among all seropositive cases, 30% are asymptomatic. Our strategy enables an accurate individual-level and multiplexed assessment of antibodies in home-sampled blood, assisting our understanding about the undiagnosed seroprevalence and diversity of the immune response against the coronavirus.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by a chronic inflammatory process often associated with comorbid asthma. In this study, we analyzed the transcriptomes of single T helper (TH) cells from nasal polyps of patients with CRSwNP and validated these findings using multiparameter flow cytometry. Polyp tissue contained suppressive T regulatory (Treg) cells, TH2 cells, type 2 innate lymphoid cells, and three transcriptionally distinct subsets of cytotoxic CD4+ T cells (CD4+ CTL). GATA3 expression was a feature of polyp Treg cells, whereas TH2 cells highly expressed TCN1, CD200R, and HPGDS and were enriched for genes involved in lipid metabolism. Only a portion of polyp TH2 cells expressed the prostaglandin D2 receptor CRTH2, whereas a subpopulation of CD109+CRTH2− TH2 cells expressed mRNA for common inhibitor receptors including LAG3 and TIM3 and produced IL-10. Together, we resolved the complexity of TH cells in patients with CRSwNP, identifying several distinct clusters of CD4+ CTL and a population of CD109+CRTH2− TH2 cells with putative regulatory potential.

Structured abstractObjectivesAs Sweden did not enforce social lockdown in response to the pandemic, it is critical to establish seropositivity to SARS-CoV-2 in healthy, active adults – here represented by blood donors and pregnant women. Random sampling was carried out in Stockholm, the country’s most populous region, and the study ran from virus emergence (March 2020) until the end of 2020, shortly prior to the first round of vaccinations, allowing for an estimate of population seropositivity in response to natural infection.DesignIn this cross-sectional prospective study, otherwise-healthy blood donors (n=2,100) and pregnant women (n=2,000) were sampled at random for consecutive weeks (at three intervals) between 14th March and 11th December 2020. Sera from all participants and a large cohort of historical controls (n=595) were screened for IgG responses against the SARS-CoV-2 spike (S) trimer and the receptor-binding domain (RBD). As a complement to standard approaches to analyze the data, a probabilistic Bayesian approach that assigns likelihood of past infection was used to analyze the population data. The study was carried out in accordance with Swedish Ethical Review Authority registration no. 2020-01807.SettingHealthy participant samples were selected from their respective pools at random through the Karolinska University Hospital.ParticipantsNone of the participants were symptomatic at the time of sampling and none had previously been hospitalized for COVID-19. No additional metadata was available from the samples.ResultsBlood donors and pregnant women showed a similar seroprevalence. After a steep rise at the start of the pandemic, the seroprevalence trajectory increased steadily in approach to the winter second-wave of infections, approaching 15% of all individuals surveyed by 11th December 2020. Importantly, 96% of antibody-positive healthy donors screened (n=56) developed neutralizing antibody responses at titers comparable to or higher than those observed in clinical trials of SARS-CoV-2 spike mRNA vaccination, supporting that mild infection engenders a competent B cell response.ConclusionsIn agreement with currently rising COVID-19 cases and ICU occupancy during a second winter wave of infections, these data demonstrate that the metropolitan Stockholm area was far from herd immunity nine months after the outbreak, with approximately one-in-six persons in the examined cohort seropositive for SARS-CoV-2.General abstractPublic health strategies to contain the pandemic continue to vary markedly across the world. In Sweden, compared to most advanced economies, social restrictions have primarily relied upon voluntary adherence to a set of recommendations and strict lockdowns have not been enforced. To better understand the development of humoral immunity to SARS-CoV-2 in the Stockholm population before the start of mass vaccinations, healthy blood donors and pregnant women (n=4,100) were sampled at random between 14th March-11th December 2020. All individuals (n=200/sampling week) were screened for anti-SARS-CoV-2 spike (S) trimer- and RBD-specific IgG responses with highly sensitive and specific ELISA assays, and the results were compared with those from historical controls (n=595). Data were modelled using a probabilistic Bayesian framework that considered individual responses to both antigens. We found that after a steep rise at the start of the pandemic, the seroprevalence trajectory increased steadily in approach to the winter second-wave of infections, approaching 15% of all individuals surveyed by 11th December. In agreement with the high transmission rate observed in the Stockholm area, seroprevalence in this cohort of active adults increased during the 9 months from the start of the outbreak, but was far from that required for herd immunity at the end of 2020.

Serological studies are critical for understanding pathogen−specific immune responses and informing public health measures (1,2). By developing highly sensitive and specific trimeric spike (S)−based antibody tests, we report IgM, IgG and IgA responses to SARS−CoV−2 in COVID−19 patients (n=105) representing different categories of disease severity. All patients surveyed were IgG positive against S. Elevated anti−SARS−CoV−2 antibody levels were associated with hospitalization, with IgA titers, increased circulating IL−6 and strong neutralizing responses indicative of intensive care status. Antibody−positive blood donors and pregnant women sampled during the pandemic in Stockholm, Sweden (weeks 14−25), displayed on average lower titers and weaker neutralizing responses compared to patients; however, inter−individual anti−viral IgG titers differed up to 1,000−fold. To provide more accurate estimates of seroprevalence, given the frequency of weak responders and the limitations associated with the dichotomization of a continuous variable (3,4), we used a Bayesian approach to assign likelihood of past infection without setting an assay cut−off. Analysis of blood donors (n=1,000) and pregnant women (n=900) sampled weekly demonstrated SARS-CoV-2-specific IgG in 7.2% (95% Bayesian CI [5.1−9.5]) of individuals two months after the peak of spring 2020 COVID−19 deaths. Seroprevalence in these otherwise healthy cohorts increased steeply before beginning to level−off, following the same trajectory as the Stockholm region deaths over this time period.