SARS-CoV-2 lineage-specific disease symptoms and disease severity in Sao Caetano do Sul city, Brazil (preprint)

Background: The city of Sao Caetano do Sul, Brazil, established a web-based platform to provide primary care to suspected COVID-19 patients, integrating clinical and demographic data and sample metadata. Here we describe lineage-specific spatiotemporal dynamics of infections, clinical symptoms, and disease severity during the first year of the epidemic. Methods: We selected and sequenced 879 PCR+ swab samples (8% of all reported cases), obtaining a spatially and temporally representative set of sequences. Daily lineage-specific prevalence was estimating using a moving-window approach, allowing inference of cumulative cases and symptom probability stratified by lineage using integrated data from the platform. Results: Most infections were caused by B.1.1.28 (41.3%), followed by Gamma (31.7%), Zeta (9.6%) and B1.1.33 (9.0%). Gamma and Zeta were associated with larger prevalence of dyspnoea (respectively 81.3% and 78.5%) and persistent fever (84.7% and 61.1%) compared to B.1.1.28 and B.1.1.33. Ageusia, anosmia, and coryza were respectively 18.9%, 20.3% and 17.8% less commonly caused by Gamma, while altered mental status was 108.9% more common in Zeta. Case incidence was spatially heterogeneous and larger in poorer and younger districts. Discussion: Our study demonstrates that Gamma was associated with more severe disease, emphasising the role of its increased disease severity in the heightened mortality levels in Brazil.

The Brazilian COVID-19 vaccination campaign: A modelling analysis of socio-demographic factors on uptake (preprint)

The COVID-19 pandemic has caused over half a million deaths in Brazil, and public healthcare nearly collapsed. Vaccination differs between states and demographics. Dose shortages delayed access. In this cross-sectional study, data were retrieved from the Brazilian Ministry of Health databases published since 17 January 2021, respectively. We developed a campaign optimality index to characterise inequality in vaccination access caused by age due to premature vaccination towards younger populations before older and vulnerable populations were fully vaccinated. We assessed geographical inequalities in full vaccination coverage and dose by age, sex, race, and socioeconomic status. Generalised linear regression was used to investigate the risk of death and hospitalisation by age group, socioeconomic status, and vaccination coverage. Vaccination coverage is higher in the wealthier South and Southeast. Men, people of colour, and low-income groups were more likely to be only partially vaccinated due to missing or delaying a second dose. Vaccination started prematurely for age groups under 50 years and may have hindered uptake of older age groups. Vaccination coverage was associated with a lower risk of death, especially in older age groups (OR: 10.5-34.8, 95% CI: (10.2, 35.9)). Risk of hospitalisation was greater in areas with higher vaccination rates due to higher access to care and reporting. Vaccination inequality persists between states, age and demographic groups despite increasing uptake. The association between hospitalisation rates and vaccination is attributed to preferential delivery to areas of greater transmission and access to healthcare.

Scale-free dynamics of Covid-19 in a Brazilian city (preprint)

Mathematical models can provide insights into the control of pandemic COVID-19, which remains a global priority. The dynamics of directly-transmitted infectious diseases, such as COVID-19, are usually described by compartmental models where individuals are classified as susceptible, infected and removed. These SIR models typically assume homogenous transmission of infection, even in large populations, a simplification that is convenient but inconsistent with observations. Here we use original data on the dynamics of COVID-19 spread in a Brazilian city to investigate the structure of the transmission network. We find that transmission can be described by a network in which each infectious individual has a small number of susceptible contacts, of the order of 2-5, which is independent of total population size. Compared with standard models of homogenous mixing, this scale-free, fractal infection process gives a better description of COVID-19 dynamics through time. In addition, the contact process explains the geographically localized clusters of disease seen in this Brazilian city. Our scale-free model can help refine criteria for physical and social distancing in order to more effectively mitigate the spread of COVID-19. We propose that scale-free COVID-19 dynamics could be a widespread phenomenon, a topic for further investigation.

Reinfection by the SARS-CoV-2 P.1 variant in blood donors in Manaus, Brazil (preprint)

The city of Manaus, north Brazil, was stricken by a severe epidemic of SARS-Cov-2 in March 2020, reaching a seroprevalence of 76% by October 2020. Nevertheless, in late November an abrupt increase in hospitalizations and deaths hit Manaus, causing higher number of deaths compared to the first epidemic wave. It has been hypothesized that virus lineages circulating in the second wave, namely the P.1 variant of concern first detected in early December in Manaus, could be better at evading immunity generated in response to previous infection with other lineages. In order to estimate the reinfection rate during the resurgence of SARS-CoV-2 in Manaus, we tested serial samples from 238 unvaccinated repeat blood donors using a SARS-CoV-2 anti-N IgG chemiluminescence microparticle assay. Blood donors were divided into six groups that reflected the inferred sequence of infection and reinfection with non-P.1 and P.1 variants. We assumed that reinfections induce a recrudescence (or boosting) of plasma anti-N IgG antibody levels, yielding a V-shaped time series of antibody reactivity levels. We infer that 16.9% (95% CI [9.48%, 28.5%]) of all presumed P.1 infections that were observed in 2021 were reinfections. If we also include cases of probable or possible reinfections (defined by considering the time period when the antibody levels are expected to grow after recovery and the range of half-lives for antibody waning after seroconversion), these percentages increase respectively to 25.8% (95% CI [16.7%, 37.4%]), and 31.0% (95% CI [21.4%, 42.5%]). Our data suggest that reinfection due to P.1 is common and more frequent than what has been detected by traditional epidemiologic, molecular and genomic surveillance of clinical cases.

Tracking the emergence of disparities in the subnational spread of COVID-19 in Brazil using an online application for real-time data visualisation. (preprint)

Brazil is one of the countries worst affected by the COVID-19 pandemic. We have developed CLIC-Brazil an online application for the real-time visualisation of COVID-19 data in Brazil at the municipality level. In the app, case and death data are standardised to allow comparisons to be made between places and over time. Estimates of Rt , a measure of the rate of propagation of the epidemic, over time are also made. Using data from the app, regression analyses identified factors associated with; the rate of initial spread, early epidemic intensity and predictions of the likelihood of occurrence of new incidence maxima. Municipalities with higher metrics of social development experienced earlier onset and faster growing epidemics, although space and time were the predominant predictive factors. Differences in the initial epidemic intensity (mean Rt ) were largely driven by geographic location and the date of local onset. This study demonstrates that by monitoring, standardising and analysing the development of an epidemic at a local level, insights can be gained into spatial and temporal heterogeneities.

Performance of vaccination with CoronaVac in a cohort of healthcare workers (HCW) - preliminary report (preprint)

Background: CoronaVac, a vaccine containing inactivated SARS-CoV-2, demonstrated efficacy of 50.39% 14 days or more after the 2nd dose. The objective of this study is to report the occurrence of symptomatic COVID-19 in a cohort of HCW vaccinated with CoronaVac and to estimate its effectiveness. Methods: CoronaVac was given to HCWs in Hospital das Clinicas on 18-21 January, 2021 (epi week 3) (22,402 HCWs), and on 14-16 February, 2021 (epi week 7) (21,652 HCWs). Weekly cases of symptomatic COVID-19 were evaluated. Using the period from 2020 epi week 24 through 2021 epi week 2 (before vaccination), a Poisson regression was fit to model the HCWs with COVID-19 of the hospital, and the officially reported cases in the city of Sao Paulo. The predicted numbers of cases among HCWs for 2021 epi weeks 3-12 were then compared to the observed numbers of cases (after vaccination). Effectiveness was estimated for weeks 9-12 (2 to 5 weeks after the 2nd dose). 142 samples after vaccination were evaluated for SARS-CoV-2 variants of concern. Results: Since the 1st dose there were 380 HCW diagnosed with COVID-19. On visual analysis, the number of cases of COVID-19 in the city increased sharply in 2021. The number of cases among the HCW did not follow. The estimated effectiveness 2 and 3 weeks after 2nd dose was 50.7% and 51.8%, respectively, and increased over the next 2 weeks. 67/142 samples (47%) were variants of concern, mostly P1 (57). Conclusion: Coronavac is effective in preventing COVID-19.

Social and racial inequalities in COVID-19 risk of hospitalisation and death across Sao Paulo state, Brazil (preprint)

BackgroundLittle evidence exists on the differential health effects of COVID-19 on disadvantaged population groups. Here we characterise the differential risk of hospitalisation and death in Sao Paulo state, Brazil and show how vulnerability to COVID-19 is shaped by socioeconomic inequalities. MethodsWe conducted a cross-sectional study using hospitalised severe acute respiratory infections (SARI) notified from March to August 2020, in the Sistema de Monitoramento Inteligente de Sao Paulo (SIMI-SP) database. We examined the risk of hospitalisation and death by race and socioeconomic status using multiple datasets for individual-level and spatio-temporal analyses. We explained these inequalities according to differences in daily mobility from mobile phone data, teleworking behaviour, and comorbidities. FindingsThroughout the study period, patients living in the 40% poorest areas were more likely to die when compared to patients living in the 5% wealthiest areas (OR: 1{middle dot}60, 95% CI: 1{middle dot}48 - 1{middle dot}74) and were more likely to be hospitalised between April and July, 2020 (OR: 1{middle dot}08, 95% CI: 1{middle dot}04 - 1{middle dot}12). Black and Pardo individuals were more likely to be hospitalised when compared to White individuals (OR: 1{middle dot}37, 95% CI: 1{middle dot}32 - 1{middle dot}41; OR: 1{middle dot}23, 95% CI: 1{middle dot}21 - 1{middle dot}25, respectively), and were more likely to die (OR: 1{middle dot}14, 95% CI: 1{middle dot}07 - 1{middle dot}21; 1{middle dot}09, 95% CI: 1{middle dot}05 - 1{middle dot}13, respectively). InterpretationLow-income and Black and Pardo communities are more likely to die with COVID-19. This is associated with differential access to healthcare, adherence to social distancing, and the higher prevalence of comorbidities. FundingThis project was supported by a Medical Research Council-Sao Paulo Research Foundation (FAPESP) CADDE partnership award (MR/S0195/1 and FAPESP 18/14389-0) (http://caddecentre.org/). This work received funding from the U.K. Medical Research Council under a concordat with the U.K. Department for International Development.

COVID-19 herd immunity in the Brazilian Amazon (preprint)

The herd immunity threshold is the proportion of a population that must be immune to an infectious disease, either by natural infection or vaccination such that, in the absence of additional preventative measures, new cases decline and the effective reproduction number falls below unity. This fundamental epidemiological parameter is still unknown for the recently-emerged COVID-19, and mathematical models have predicted very divergent results. Population studies using antibody testing to infer total cumulative infections can provide empirical evidence of the level of population immunity in severely affected areas. Here we show that the transmission of SARS-CoV-2 in Manaus, located in the Brazilian Amazon, increased quickly during March and April and declined more slowly from May to September. In June, one month following the epidemic peak, 44% of the population was seropositive for SARS-CoV-2, equating to a cumulative incidence of 52%, after correcting for the false-negative rate of the antibody test. The seroprevalence fell in July and August due to antibody waning. After correcting for this, we estimate a final epidemic size of 66%. Although non-pharmaceutical interventions, plus a change in population behavior, may have helped to limit SARS-CoV-2 transmission in Manaus, the unusually high infection rate suggests that herd immunity played a significant role in determining the size of the epidemic.

Baseline T cell immune phenotypes predict virologic and disease control upon SARS-CoV infection (preprint)

The COVID-19 pandemic has revealed that infection with SARS-CoV-2 can result in a wide range of clinical outcomes in humans, from asymptomatic or mild disease to severe disease that can require mechanical ventilation. An incomplete understanding of immune correlates of protection represents a major barrier to the design of vaccines and therapeutic approaches to prevent infection or limit disease. This deficit is largely due to the lack of prospectively collected, pre-infection samples from indiviuals that go on to become infected with SARS-CoV-2. Here, we utilized data from a screen of genetically diverse mice from the Collaborative Cross (CC) infected with SARS-CoV to determine whether circulating baseline T cell signatures are associated with a lack of viral control and severe disease upon infection. SARS-CoV infection of CC mice results in a variety of viral load trajectories and disease outcomes. Further, early control of virus in the lung correlates with an increased abundance of activated CD4 and CD8 T cells and regulatory T cells prior to infections across strains. A basal propensity of T cells to express IFNg and IL17 over TNFa also correlated with early viral control. Overall, a dysregulated, pro-inflammatory signature of circulating T cells at baseline was associated with severe disease upon infection. While future studies of human samples prior to infection with SARS-CoV-2 are required, our studies in mice with SARS-CoV serve as proof of concept that circulating T cell signatures at baseline can predict clinical and virologic outcomes upon SARS-CoV infection. Identification of basal immune predictors in humans could allow for identification of individuals at highest risk of severe clinical and virologic outcomes upon infection, who may thus most benefit from available clinical interventions to restrict infection and disease.

Comparative analysis reveals the species-specific genetic determinants of ACE2 required for SARS-CoV-2 entry (preprint)

Coronavirus interaction with viral receptor is a primary genetic determinant of host range and tissue tropism. SARS-CoV-2 utilizes ACE2 as the receptor to enter the host cell in a species-specific manner. We and others have previously shown that ACE2 orthologs from New World monkeys, koala and mouse cannot interact with SARS-CoV-2 to mediate viral entry, and this defect can be restored by humanization of the restrictive residues in New World monkey ACE2. To better understand the genetic determinants of susceptibility of ACE2 orthologs to viral entry, we compared koala and mouse ACE2 sequences with human ortholog, and identified the key residues in koala or mouse ACE2 that restrict its viral receptor activity. Humanization of these critical residues could render the capabilities of koala and mouse ACE2 to bind viral spike protein and facilitate the viral entry. Our work identifies the genetic determinant of ACE2 for SARS-CoV-2 susceptibility, and a single mutation could restore the mouse ACE2 receptor activity, providing a potential avenue for the development of mouse model of SARS-CoV-2.

Cryo-EM structure of S-Trimer, a subunit vaccine candidate for COVID-19 (preprint)

Less than a year after its emergence, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected over 22 million people worldwide with a death toll approaching 1 million. Vaccination remains the best hope to ultimately put this pandemic to an end. Here, using Trimer-Tag technology, we produced both wild-type (WT) and furin site mutant (MT) S-Trimers for COVID-19 vaccine studies. Cryo-EM structures of the WT and MT S-Trimers, determined at 3.2 Angstrom and 2.6 Angstrom respectively, revealed that both antigens adopt a tightly closed conformation and their structures are essentially identical to that of the previously solved full-length WT S protein in detergent. These results validate Trimer-Tag as a platform technology in production of metastable WT S-Trimer as a candidate for COVID-19 subunit vaccine.

SARS-CoV-2 Nsp1 suppresses host but not viral translation through a bipartite mechanism (preprint)

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a highly contagious virus that underlies the current COVID-19 pandemic. SARS-CoV-2 is thought to disable various features of host immunity and cellular defense. The SARS-CoV-2 nonstructural protein 1 (Nsp1) is known to inhibit host protein translation and could be a target for antiviral therapy against COVID-19. However, how SARS-CoV-2 circumvents this translational blockage for the production of its own proteins is an open question. Here, we report a bipartite mechanism of SARS-CoV-2 Nsp1 which operates by: (1) hijacking the host ribosome via direct interaction of its C-terminal domain (CT) with the 40S ribosomal subunit and (2) specifically lifting this inhibition for SARS-CoV-2 via a direct interaction of its N-terminal domain (NT) with the 5 untranslated region (5 UTR) of SARS-CoV-2 mRNA. We show that while Nsp1-CT is sufficient for binding to 40S and inhibition of host protein translation, the 5 UTR of SARS-CoV-2 mRNA removes this inhibition by binding to Nsp1-NT, suggesting that the Nsp1-NT-UTR interaction is incompatible with the Nsp1-CT-40S interaction. Indeed, lengthening the linker between Nsp1-NT and Nsp1-CT of Nsp1 progressively reduced the ability of SARS-CoV-2 5 UTR to escape the translational inhibition, supporting that the incompatibility is likely steric in nature. The short SL1 region of the 5 UTR is required for viral mRNA translation in the presence of Nsp1. Thus, our data provide a comprehensive view on how Nsp1 switches infected cells from host mRNA translation to SARS-CoV-2 mRNA translation, and that Nsp1 and 5 UTR may be targeted for anti-COVID-19 therapeutics.

An immunodominance hierarchy exists in CD8+ T cell responses to HLA-A*02:01-restricted epitopes identified from the non-structural polyprotein 1a of SARS-CoV-2. (preprint)

COVID-19 vaccines are being rapidly developed and human trials are underway. Almost all of these vaccines have been designed to induce antibodies targeting spike protein of SARS-CoV-2 in expectation of neutralizing activities. However, non-neutralizing antibodies are at risk of causing antibody-dependent enhancement. Further, the longevity of SARS-CoV-2-specific antibodies is very short. Therefore, in addition to antibody-induced vaccines, novel vaccines on the basis of SARS-CoV-2-specific cytotoxic T lymphocytes (CTLs) should be considered in the vaccine development. Here, we attempted to identify HLA-A*02:01-restricted CTL epitopes derived from the non-structural polyprotein 1a of SARS-CoV-2. Eighty-two peptides were firstly predicted as epitope candidates on bioinformatics. Fifty-four in 82 peptides showed high or medium binding affinities to HLA-A*02:01. HLA-A*02:01 transgenic mice were then immunized with each of the 54 peptides encapsulated into liposomes. The intracellular cytokine staining assay revealed that 18 out of 54 peptides were CTL epitopes because of the induction of IFN-{gamma}-producing CD8+ T cells. In the 18 peptides, 10 peptides were chosen for the following analyses because of their high responses. To identify dominant CTL epitopes, mice were immunized with liposomes containing the mixture of the 10 peptides. Some peptides were shown to be statistically predominant over the other peptides. Surprisingly, all mice immunized with the liposomal 10 peptide mixture did not show the same reaction pattern to the 10 peptides. There were three pattern types that varied sequentially, suggesting the existence of an immunodominance hierarchy, which may provide us more variations in the epitope selection for designing CTL-based COVID-19 vaccines. ImportanceFor the development of vaccines based on SARS-CoV-2-specific cytotoxic T lymphocytes (CTLs), we attempted to identify HLA-A*02:01-restricted CTL epitopes derived from the non-structural polyprotein 1a of SARS-CoV-2. Out of 82 peptides predicted on bioinformatics, 54 peptides showed good binding affinities to HLA-A*02:01. Using HLA-A*02:01 transgenic mice, 18 in 54 peptides were found to be CTL epitopes in the intracellular cytokine staining assay. Out of 18 peptides, 10 peptides were chosen for the following analyses because of their high responses. To identify dominant epitopes, mice were immunized with liposomes containing the mixture of the 10 peptides. Some peptides were shown to be statistically predominant. Surprisingly, all immunized mice did not show the same reaction pattern to the 10 peptides. There were three pattern types that varied sequentially, suggesting the existence of an immunodominance hierarchy, which may provide us more variations in the epitope selection for designing CTL-based COVID-19 vaccines.

A primary care approach to the COVID-19 pandemic: clinical features and natural history of 2,073 suspected cases in the Corona Sao Caetano programme, Sao Paulo, Brazil (preprint)

Background: Despite most cases not requiring hospital care, there are limited community-based clinical data on COVID-19. Methods and findings: The Corona Sao Caetano program is a primary care initiative offering COVID-19 care to all residents of Sao Caetano do Sul, Brazil. After triage of potentially severe cases, consecutive patients presenting between 13th April and 13th May 2020 were tested at home with SARS-CoV-2 reverse transcriptase (RT) PCR; positive patients were followed up for 14 days. RT-PCR-negative patients were offered SARS-CoV-2 serology. We describe the clinical features, virology and natural history of this prospective population-based cohort. Of 2,073 suspected COVID-19 cases, 1,583 (76.4%) were tested by RT-PCR, of whom 444 (28.0%, 95%CI: 25.9% - 30.3%) were positive; 604/1,136 (53%) RT-PCR-negative patients underwent serology, of whom 52 (8.6%) tested SARS-CoV-2 seropositive. The most common symptoms of COVID-19 were cough, fatigue, myalgia and headache; whereas self-reported fever, anosmia, and ageusia were most associated with a positive COVID-19 diagnosis. RT-PCR cycle thresholds were lower in men, older patients, those with fever and arthralgia, and around symptom onset. The rates of hospitalization and death among 444 RT-PCR-positive cases were 6.7% and 0.7%, respectively, with older age and obesity more frequent in the hospitalized group. Conclusions: COVID-19 presents similarly to other mild respiratory disease in primary care. Some symptoms assist the differential diagnosis. Most patients can be managed at home.

Routes for COVID-19 importation in Brazil (preprint)

The global outbreak caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been declared a pandemic by the WHO. As the number of imported SARS-CoV-2 cases is on the rise in Brazil, we use incidence and historical air travel data to estimate the most important routes of importation into the country.

Routes for COVID-19 importation in Brazil ;Journal of Travel Medicine ;Oxford Academic

Highlight The global outbreak caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been declared a pandemic by the WHO. As the number of imported SARS-CoV-2 cases is on the rise in Brazil, we use incidence and historical air travel data to estimate the most important routes of importation into the country.