Emerging Microbes & Infections

Influenza A(H3N2) subclade K virus was detected in Canada early in the 2025/26 influenza season, bearing an antigenic transition in the hemagglutinin (HA) glycoprotein. Analysis of 396 HA sequences from Canada showed antigenic divergence from 2025/26 influenza vaccine strains, consistent with partial mismatch. Phylodynamic analysis revealed sustained pre-vaccine transmission without clear post-vaccine expansion. Phylogenetic and phylogeographic analyses indicated interprovincial mixing within a highly connected metapopulation, highlighting the value of genomic surveillance for real-time epidemiologic inference and public health decision-making.

In the past decade, dengue fever has emerged as a major public health on Reunion Island in the Southwest Indian Ocean. During the 2018-2022 outbreak, an unusual increase in ocular complications was reported in some patients. To investigate a potential viral cause, we analysed 447 blood samples from hospitalized patients with and without ophthalmic symptoms. Genetic sequencing revealed the co-circulation of two strains of dengue virus serotype 1, both genetically linked to strains previously identified in Asia. Notably, all patients with ophthalmic symptoms were infected with viruses from a single cluster within genotype I, which harbored several unique mutations. These findings suggest that the rare ocular complications observed during this outbreak may be associated with specific viral cluster. Further laboratory studies are required to confirm this potential link.

Household transmission of EV-D68 was identified in 35 of 1040 households (3.4%) in the Pacific Northwest between 2022-2024, with an estimated secondary attack rate of 15%. Sequences from within households clustered closely with 0 to 2 pairwise nucleotide differences (median 1) between cases 6-14 days apart (median 7).

The emergence of vaccine covered serotypes causing invasive pneumococcal disease (IPD) is a serious concern worldwide. We investigated the unexpected rise of serotype 4 causing IPD primarily in non-vaccinated young adults after the COVID-19 pandemic that further spread to adults [≥] 65 years in recent years. For this purpose, we conducted a retrospective study of serotype 4 IPD cases (n=827) reported in Spain between 2009 and 2024. Whole-genome sequencing was performed to assess clonal lineages and phylogenetic relationships. Clinical and epidemiological data were compared between serotype 4 and all other serotypes causing IPD. Epidemiological and genomic analysis confirmed that the rise started as an abrupt cluster of IPD cases in Seville (Andalusia) in the year 2022 due to the ST15063 within GPSC12 lineage. This outbreak initially caused pneumonia episodes that required hospitalization in young individuals associated with high rates of tobacco smoking, alcohol, and inhaled drugs such as cannabis and cocaine, followed by a general distribution pattern throughout the country in the following years, affecting the elderly population. Experimental studies to evaluate potential underlying mechanisms confirmed that ST15063 serotype 4 strains displayed enhanced infection rates of human lung cells that significantly increased in the presence of cigarette smoke exposure and by influenza H3N2 virus coinfection, but not with H1N1. These findings highlight the need for targeted vaccination strategies not only against pneumococcus but also against respiratory viruses such as influenza, RSV and COVID-19 and demonstrate the importance of molecular surveillance to establish effective interventions in high-risk populations.

The rapid diagnosis of Campylobacter infections is important for the management of infectious gastroenteritis. Although stool culture is considered the gold standard, its sensitivity is limited and it requires prolonged incubation times. We performed a prospective multicenter study at nine healthcare facilities in Japan to evaluate a Campylobacter rapid antigen test using stool specimens between March 2024 and August 2025. Patients with suspected infectious gastroenteritis were consecutively enrolled and tested using QuickNavi-Campylobacter and compared with the FilmArray Gastrointestinal Panel as the reference method. Discordant results were further evaluated by culturing and additional PCR assays. In total, 410 patients were included in the final analysis. The positive, negative, and total concordance rates between QuickNavi-Campylobacter and FilmArray Gastrointestinal Panel were 79%, 99%, and 93%, respectively. The positive concordance rate decreased in specimens collected [≥] 6 days after the onset of symptoms (50%). QuickNavi-Campylobacter demonstrated relatively good concordance with the FilmArray Gastrointestinal Panel in a real-world multicenter setting. These results suggest that this rapid antigen test may be particularly useful for the early diagnosis of suspected campylobacteriosis.

BackgroundThere is a need for synthetic peptide-based serologic assays that exploit avidity to replace whole antigens while enabling low-cost diagnostics in resource-limited settings. ObjectiveTo evaluate the diagnostic accuracy of a polymeric peptide-based ELISA leveraging avidity to enhance signal. MethodA 15-member SARS-CoV-2 peptide library corresponding to multiple epitope clusters and proteins was screened by indirect ELISA using pooled sera from RT-PCR-confirmed COVID-19 patients to identify peptides with possible diagnostic utility. The identified lead candidate, S559, possessed terminal cysteine-substitution to allow disulfide polymerization, and the resulting avidity gain was evaluated by comparing the apparent dissociation constant (KDapp) before and after depolymerization with N-acetylcysteine. The performance of an optimized ELISA using S559 was evaluated on 1,222 prospectively collected COVID-19 serum samples and 218 biobanked pre-COVID control serum samples. ResultsPolymeric S559 with a KDapp of 29.26 nM-1was demonstrated to have a 218% avidity gain relative to the completely depolymerized form. At pre-defined thresholds, the optimized S559 ELISA has a sensitivity and specificity of 83.39% (95%CI: 81.18% and 85.43%) and 96.79% (95%CI: 93.50% and 98.70%), respectively. At post hoc thresholds determined by Youden index, sensitivity and specificity reached 95.01 (95% CI: 93.63% - 96.16%) and 100.00% (95% CI: 98.32% - 100.00%), respectively. ConclusionHomomultivalent epitope presentation using polymeric S559 allows a highly specific immunoassay using human sera that may have important value in detecting antibodies, whether for diagnosing infection, confirming vaccination status or conducting surveillance.

Strengthening in-country sequencing capacity generated 28 Lassa virus genomes from human clinical cases, expanding our knowledge of Lassa fever in Guinea. Phylogeographic analysis revealed cross-border exchange between Liberia and the NZerekore region, and a Sierra Leone introduction into the Gueckedou area. Enhanced genomic surveillance is crucial to guide future public health actions.

RIG-I is a cytosolic immune receptor that provides the first line of defense by detecting viral RNA and triggering antiviral responses. Its physiological role in humans remains unclear, as no patients with complete RIG-I deficiency have yet been reported. We identified a critically ill COVID-19 patient with severe RIG-I deficiency caused by heterozygous RIG-I G731R, a novel dominant loss-of-function variant. The G731R mutation in helicase motif VI disrupts the arginine finger, impairing the ATPase activity of RIG-I, but not its RNA-binding ability. However, viral RNA binding fails to expose the signaling domains, thereby impairing the IFN-{beta} response of G731R. Instead, G731R competes with wild-type RIG-I, exerting a dominant negative effect. The loss-of-function is caused by bulky-charged substitutions at G731, as alanine or leucine substitution results in an unexpected gain-of-function phenotype. These findings highlight the importance of uncompromised RIG-I function for human antiviral immunity and the pleiotropic effects of single mutations.

BackgroundDengue fever is a major neglected tropical disease with a rapidly rising global burden, and localized outbreaks are increasingly reported in southern subtropical China. Fujian Province, a coastal subtropical region with favorable ecological conditions for Aedes albopictus breeding and frequent cross-border exchanges with dengue-endemic areas, has had continuous local dengue cases for over a decade, raising concerns about the establishment of a stable natural endemic focus. Sustained local dengue transmission is defined by four core criteria, but no systematic assessment of these criteria has been conducted for Fujian using long-term multi-dimensional surveillance data. We aimed to evaluate whether a natural endemic focus for sustained local dengue transmission has been established in Fujian Province from 2014 to 2024 using four core evidence dimensions. MethodsWe extracted data on imported and locally acquired dengue cases in Fujian from 2014 to 2024 from Chinas National Notifiable Disease Reporting System (NNDRS). Serological surveillance for dengue IgG antibodies and virological surveillance for dengue virus in Aedes albopictus were conducted at seven sentinel sites. The study period was stratified into three phases based on the impact of COVID-19 non-pharmacological interventions: pre-pandemic (2014-2019), pandemic(2020-2022), and post-pandemic(2023-2024). Descriptive epidemiological analysis and data visualization were performed using R software (version 4.4.1), with t-tests for continuous variables and {chi}{superscript 2} tests for categorical variables. ResultsA total of 3,606 dengue cases were reported in Fujian during the study period, including 1,229 imported and 2,377 locally acquired cases. Key findings were as follows: (1) Temporal distribution: Local dengue transmission was completely interrupted during the 2020-2022 COVID-19 pandemic (0 local cases, only 26 imported cases), and resumed at a low level in 2023-2024 (160 local cases). (2) Serology: The overall population dengue IgG antibody positivity rate was 4.2% (66/15,736), with no statistically significant difference between pre-epidemic (3.8%, 30/7,835) and post-epidemic seasons (4.5%, 36/7,901; P=0.48), and no year with a positivity rate exceeding 10%. (3) Vector surveillance: Only one dengue virus-positive sample was detected among 385,000 Aedes albopictus mosquitoes collected during routine surveillance (Taijiang District, Fuzhou, October 2017), with no viral nucleic acid detected in all other samples. (4) Age distribution: The mean age of locally acquired cases (46.1{+/-}19.8 years) was significantly higher than that of imported cases (35.8{+/-}11.2 years, P<0.001), and local cases were concentrated in the middle-aged group (40-60 years) with no child-dominant pattern observed. ConclusionsFujian Province has not established a stable natural endemic focus for sustained local dengue transmission, and imported cases are the primary driver of local outbreaks in the region. Strengthened surveillance and early management of imported cases, integrated vector control targeting Aedes albopictus, and targeted public health education are critical and essential strategies to prevent the establishment of a dengue natural endemic focus in Fujian and other subtropical coastal regions with similar epidemiological characteristics. Author SummaryDengue fever is a rapidly spreading neglected tropical disease worldwide, and southern China faces persistent threats of local transmission due to favorable ecological conditions for mosquito breeding and frequent cross-border travel. Fujian Province, a subtropical coastal region in southeastern China, has reported annual local dengue cases for over a decade, raising public health concerns about the potential establishment of a stable natural endemic focus--where the virus circulates sustainably without relying on imported cases. To address this critical question, we conducted a comprehensive 11-year assessment (2014-2024) of dengue transmission in Fujian using four key evidence dimensions defined for identifying dengue endemic foci: the continuity of local cases independent of imported sources, population antibody levels, dengue virus detection in local mosquitoes (Aedes albopictus), and the age distribution of infected patients. We also leveraged the COVID-19 pandemic(2020-2022) as a unique natural experiment, during which strict travel restrictions drastically reduced imported dengue cases, to test whether local transmission could persist on its own. Our findings showed that local dengue transmission in Fujian completely stopped during the COVID-19 pandemic and only resumed when cross-border travel and imported cases recovered, confirming local transmission is entirely dependent on imported virus sources. Additionally, the local population had a very low dengue antibody positivity rate (4.2%), dengue virus was detected in only one mosquito sample over 11 years of surveillance, and local cases were concentrated in middle-aged adults (not children--the typical group affected in endemic areas). Together, these results confirm that Fujian Province has not established a stable natural endemic focus for dengue fever. While no endemic focus exists yet, Fujian remains at high risk of imported-driven local outbreaks due to its climate and cross-border exchanges. Our study highlights three critical strategies to prevent the future establishment of a dengue endemic focus in Fujian and other similar subtropical coastal regions: strengthening surveillance and early response for imported dengue cases, implementing targeted mosquito control measures during peak transmission seasons, and conducting public health education to raise awareness of dengue prevention. These evidence-based interventions are key to blocking the formation of sustained local dengue transmission and protecting regional population health.

Herpes simplex virus (HSV) is an endemic pathogen, infecting most adults world-wide. HSV infection can cause a wide spectrum of disease outcomes, ranging from asymptomatic infection or mild lesions to rare cases of infectious keratitis, encephalitis, and death. HSV genome sequences have been shown to differ between individual patients, as well as within individuals. To date, the vast majority of publicly available HSV genomic data has come from Europe and North America. Our current understanding of these patterns are missing data from under-sampled populations, particularly in South America, Africa, and Asia. Also missing have been HSV samples from non-industrial (e.g., agricultural, pastoral) populations, for which the natural environment plays a large role in health and disease dynamics. In this study, we capitalized on Whatman FTA card stabilization of DNA to develop a procedure for capturing oral and genital swabs from a geographically isolated pastoralist population in a desert region of northern Namibia. These are the first data to document HSV diversity in this type of remote setting. These are also the first HSV genomes from Namibia. These approaches may prove useful in broadening the accessibility of viral detection for these chronic pathogens, help improve diagnostics, and raise public health awareness about the burden of these pathogens in under-served populations.

Topiramate (TPM) is approved for seizures and migraine prophylaxis and is used off-label for several neuropsychiatric conditions. The available dosage forms, including tablets and sprinkle capsules, are unsuitable for patients who may be unable to take medicine orally. The resulting potential treatment interruptions could have untoward consequences and underscores the importance of developing a parenteral formulation. In this study, we developed a population pharmacokinetic model of a novel, intravenous TPM formulation using data from a study in patients with epilepsy or migraine receiving a single intravenous dose of stable-labeled TPM. In total, 246 TPM concentrations from 20 adult patients were included for model development. A three-compartment pharmacokinetic model with linear elimination fit the concentration-time data best. Simulations for various loading and maintenance regimens for patients with and without enzyme-inducing comedications were performed. The final estimates(95% confidence interval (CI)) for CL (L/h), V1 (L), and the peripheral volumes, V2 and V3 for a 70 kg person were 1.31(1.01 - 1.53), 9.84 (8.49 - 11.0), 39.1 (36.5 - 41.8)L, and 9.01 (6.41 - 44.3) respectively. The use of enzyme-inducing co-medication was the only significant covariate, associated with a 63% increase in clearance .Goodness-of-fit plots and visual predictive checks indicate satisfactory model performance and prediction. The simulation results indicate that adjusting doses for patients receiving IV TPM can mitigate the changes in plasma TPM concentrations resulting from enzyme induction. This population pharmacokinetic model for intravenous topiramate can inform dosing decisions for patients with epilepsy when used as either initiation or bridging therapy.

Genomic surveillance of influenza viruses informs vaccine strain selection and evolutionary forecasting. Sequencing efforts vary widely across U.S. states, which raises concerns about spatial sampling bias. We evaluated how well 10,958 influenza virus genomes sampled by our group in Michigan captured the genetic diversity in 34,743 genomes circulating nationally from the 2021/22 through 2024/25 seasons. We defined seasonal hemagglutinin haplotypes and tracked their detection across states. A small number of haplotypes dominated each season, and Michigan detected all major haplotypes, even under substantial downsampling. Detection delays were primarily driven by haplotype frequency rather than geographic factors. Comparisons across states showed that higher sequencing effort improved coverage and detection timeliness, with diminishing returns at higher volumes. Rarefaction analysis confirmed that relatively few sequences were needed to capture 95% of national haplotype diversity. These findings suggest that intensive sequencing in a single well-sampled location can be broadly representative of national influenza diversity. One sentence summaryDense influenza genomic sequencing from a single U.S. state captured nearly all nationally circulating haplotype diversity, with detection timeliness primarily driven by sequencing effort and haplotype frequency.

The measles outbreak in Jalisco, Mexico (January-February 2026) experienced vigorous sustained transmission with an exponential growth rate = 0.10 (95% CI: 0.10-0.11) per day, doubling time = 6.3 days (95% CI: 6.3-6.9), yielding the effective reproduction number at 3.34 (95% CI: 3.16-3.54), with elevated incidence among infants and young adults.

IntroductionDengue has been prevalent in a regular fashion in Bangladesh and Chattogram for the last 6-7 years and is showing some serotype twisting. So, the objectives of the present study were to explore the burden of dengue serotypes in Chattogram. MethodsIn this study, 223 Dengue RT-PCR positive patients were evaluated for serotyping. Gender and age group, along with cycle threshold (CT) values, were also collected. Data after collection were compiled, analyzed, and plotted in Microsoft Excel and GraphPad Prism 10.4. Ethical clearance was taken to conduct the study. ResultsAmong 223 patients analyzed, males and females were found near equal (113 and 110). Middle-aged patients were more than the extremes of age. The mean {+/-} SD of age was 33.55 {+/-} 13.67 years. Regarding serotype distributions, isolated Den 1, Den 2 and Den 3 were found 1.3%, 73.1% and 6.7%, respectively. Concurrent infections with multiple serotypes were observed in several patients, most notably the Den 2 and Den 3 combination, which accounted for 14.3% (n=32) of the cases. Other co-infections were less frequent: the Den 1 and Den 2 pairing appeared in 3.6% (n=8) of the cohort, while triple-serotype infections (Den 1, 2, and 3) and Den 3/Den 4 pairings were rare, each occurring in only 0.4% of patients. Statistical analysis of CT values revealed no significant sex-based differences for Den 2 and Den 3. However, significant variations in CT values were observed when comparing Den 1 against both Den 2 and Den 3 (p < 0.05). In contrast, the difference between Den 2 and Den 3 Ct values remained statistically insignificant. ConclusionIn the year 2025, Dengue serotypes 2 and 3 were found to be the most prevalent, both in isolated or in combinations and Den 1 and Den 4 were found minimum. Exposure to multiple serotypes and twisting from one serotype to another might influence the dengue outcome in future, which needs further exploration.

An H3N2 variant, named subclade K, continues to circulate widely during the 2025-2026 influenza season. This virus possesses a hemagglutinin (HA) protein that has eleven substitutions relative to the HA of the Northern Hemisphere 2025-2026 H3N2 vaccine strain. Many of these substitutions are in epitopes in well-characterized HA antigenic sites. Despite this, interim vaccine effectiveness studies indicate that the 2025-2026 influenza vaccine provides moderate protection against H3N2 subclade K infection. We previously reported that many individuals who received the 2025-2026 influenza vaccine produced antibodies that inhibit H3N2 subclade K virus cellular attachment. Here, we show these individuals also produced antibodies that neutralize H3N2 subclade K virus infection, and we observed a strong correlation between hemagglutination-inhibition titers and neutralizing antibody titers. We completed additional specificity studies using samples from individuals who did or did not have antibodies that cross-reacted to H3N2 subclade K viruses. Using high-throughput neutralization assays, we determined that antibodies that bound to the vaccine strain but not H3N2 subclade K viruses typically targeted antigenic site B of HA. Conversely, we found that cross-reactive neutralizing antibodies elicited by vaccination commonly targeted antigenic site A, D, and E of HA that are conserved between the vaccine strain and H3N2 subclade K viruses. Additional electron microscopy-based polyclonal epitope mapping studies confirmed that cross-reactive antibodies elicited by vaccination typically target epitopes on the side of HA. Together, our studies provide an immunological explanation of why the 2025-2026 influenza vaccine was partially effective against antigenically advance H3N2 subclade K viruses. Our data suggest that vaccine strains for subsequent seasons need to be carefully considered, since subclade K viruses have already started to acquire additional substitutions in HA antigenic sites targeted by cross-reactive antibodies.

Clade 2.3.4.4b highly pathogenic avian influenza A(H5N1) viruses continue to expand geographically and across mammalian hosts, raising concern about pandemic potential. The degree and specificity of pre-existing immunity in humans are key determinants of this risk. We analyzed hemagglutinin (HA)-and neuraminidase (NA)-specific antibody responses in 300 sera collected from adults in New York City. While HA directed binding antibodies to clade 2.3.4.4b H5 were low and hemagglutination-inhibiting antibodies were absent, we detected widespread binding and functional NA antibodies against N1 neuraminidases from clade 2.3.4.4b H5N1 viruses. Neuraminidase inhibition (NI) titers were highest against North American D1.1 genotype N1 viruses and correlated strongly with neutralizing activity, whereas HA-binding antibodies did not. An additional N-linked glycosylation site, as found in the NA of a human D1.1 isolate from British Columbia, reduced susceptibility to NI antibodies. Antibodies to N5 from H5N5 were minimal. These findings indicate that population-level immunity to clade 2.3.4.4b H5 viruses is dominated by NA-directed antibodies, with important implications for pandemic risk assessment. ImportanceUnderstanding how pre-existing human immunity shapes susceptibility to emerging influenza viruses is central to pandemic preparedness. Here, we determined that human sera contain widespread, functional antibodies targeting H5N1 neuraminidase, which correlate with virus neutralization, whereas HA directed responses are limited. We further show that acquisition of an NA glycosylation site reduces antibody inhibition, highlighting a potential pathway for immune evasion. These results identify neuraminidase-specific immunity as a major immunological barrier to severe H5N1 disease in humans and emphasize the need to incorporate NA antigenicity into influenza surveillance, risk assessment, and next-generation vaccine design.

AO_SCPLOWBSTRACTC_SCPLOWInfluenza A subclade K viruses caused high infection rates in the 2025/2026 Northern Hemisphere season, raising concerns about antigenic drift and reduced vaccine effectiveness. We measured antibody responses in matched human pre- and post-vaccination sera against a vaccine-like as well as subclade K isolates. Pre-existing immunity to subclade K variants was noted with seasonal influenza vaccination boosting titers two-fold against subclade K and three-fold against the vaccine-like strain, indicating little antigenic drift. These data contrast ferret-based predictions of marked antigenic drift.

Dengue virus (DENV), comprising four distinct serotypes (DENV-1 to DENV-4), poses a major public health challenge in tropical regions. Infection with one serotype confers long-term immunity to that serotype alone, while subsequent heterologous infections are associated with increased risk of severe disease, necessitating vaccines that induce durable, balanced immunity across all serotypes. However, achieving such balance immunity remains a central challenge for dengue vaccine development. Using passive surveillance data from Kamphaeng Phet, Thailand (2004-2022), we characterized long-term serotype circulation and contributions to clinical dengue burden in a hyperendemic setting. We observed sustained co-circulation of all four serotypes over nearly two decades, with periodic shifts in dominant serotype. Among 6,840 PCR-confirmed dengue cases, the majority of which were hospitalized, DENV-1 through DENV-4 accounted for 32.8%, 25.9%, 24.8%, and 16.5% of detected dengue cases, respectively. Compared with DENV-1, clinically-apparent DENV-2 and DENV-4 infections were more likely to represent secondary infections (odds ratio 4.94 and 5.24, respectively) and occurred at older ages, underscoring the context-dependent clinical expression of different serotypes. Together, these findings demonstrate that all four dengue serotypes contribute meaningfully to clinical disease burden in Thailand and caution against de-emphasizing individual serotypes based on transient epidemiologic patterns. These results reinforce the importance of tetravalent vaccine strategies alongside continued evaluation of vaccine performance in evolving epidemiologic settings. Author SummaryDengue is a mosquito-borne disease that infects millions of people every year. The virus consists of four serotypes. Infection with one serotype does not provide full protection against the others, and a second infection with a different serotype can increase the likelihood of severe illness. For this reason, understanding which serotypes are circulating is important for designing and evaluating effective vaccines. Our analysis of nearly two decades of surveillance data from Kamphaeng Phet, Thailand demonstrates that all four serotypes were consistently present over time, each contributing a substantial number of cases. The serotypes showed distinct age- and immunity-dependent epidemiologic patterns. No single serotype could be considered unimportant. These findings highlight the complex nature of DENV transmission in Thailand and emphasize the need for vaccines that provide protection against all four serotypes. Continuous monitoring of circulating serotypes is essential to guide vaccine development and to ensure their effectiveness in real-world settings.

Newly emerging influenza virus strains pose a constant threat as they encounter a population lacking neutralizing antibodies against the new strain. However, cross-reactive non-neutralizing antibodies (nnABs) may be present and assist in mitigating disease symptoms via various effector mechanisms, including antibody-dependent cellular cytotoxicity (ADCC). Although nnABs to influenza virus have received more attention lately, little information is available on their age-related prevalence, steady-state levels, functional properties, and changes in these parameters over time. Using longitudinal samples from adolescents, adults, and older adults, collected before and after the 2009 swine flu pandemic, we comprehensively characterized the specificity and functionality of nnAB responses against H1N1 pandemic 2009 (H1N1pdm09) virus. Remarkably, all participants exhibited cross-reactive antibodies to this virus before having encountered it through infection or vaccination, with the highest baseline levels observed in older adults. The levels of these IgG antibodies showed a strong correlation with engagement of fragment crystallizable {gamma} receptor IIIa (Fc{gamma}RIIIa) and ADCC activity, both of which were notably lower in adolescents compared to adults and older adults. Without infection or vaccination, average amounts of H1N1pdm09-reactive antibodies remained relatively stable on population level over the 5-year study period. However, on an individual level, substantial increases and decreases occurred. H1N1pdm09 infection or vaccination significantly enhanced specific antibody levels and the Fc{gamma}RIIIa-engaging capacity of these antibodies in all age groups. ADCC-mediating antibodies increased however only in adolescents, reaching the same level as observed in the adult groups. Taken together, our results demonstrate the presence of cross-reactive, non-neutralizing, functional, and boostable antibodies against a never-encountered influenza virus strain across all age groups. These antibodies can potentially contribute to protection from severe disease. Accordingly, in case of a newly emerging virus, their further enhancement by vaccination could be beneficial as an immediate protective measure before a strain-specific vaccine becomes available. Author summaryNearly everyone has contracted influenza and/or has been vaccinated against influenza several times over the years. While the antibodies raised during these earlier encounters will not prevent infection by a newly emerging influenza virus strain, they can help to protect from severe disease. Therefore, it is important to determine the prevalence and quantity of these antibodies, understand their mechanisms of action, assess their persistence over time, and examine potential age-related differences in these parameters. We studied antibody responses to the H1N1pdm09 virus in blood samples of young, adult, and older adult individuals from a large cohort study. Irrespective of age, all blood samples contained antibodies that reacted with a never-before-encountered influenza virus strain. The amounts of these antibodies were initially lower in adolescents but with time increased, reaching the same levels as observed in adults. Importantly, infection with or vaccination against the new virus strengthened the responses in all age groups. We conclude that boosting such broadly-reactive antibodies through vaccination could serve as an immediate strategy when a new virus emerges, buying critical time to develop a more specific vaccine.

BackgroundHeartland virus (HRTV) is an emerging tick-borne virus capable of causing severe illness and death. The burden of disease is likely underestimated due to limited seroprevalence studies, lack of commercially available diagnostic tests, and an overlapping clinical syndrome with more commonly diagnosed bacterial diseases such as spotted fever group rickettsiosis or ehrlichiosis. MethodsActive surveillance for Heartland virus disease was conducted at a large academic center from March to September 2024. Enrolled subjects included those who had testing sent for Ehrlichia polymerase chain reaction (PCR) along with fever and 2 of the 3 criteria: leukopenia, thrombocytopenia, and/or elevated liver function tests. Specimens with detectable RNA underwent whole genome sequencing and analysis. FindingsOver 800 specimens were received with 53 individuals meeting enrollment criteria. Among these 53, two (3.8%) had detectable HRTV RNA in whole blood during the time of Ehrlichia PCR testing. The two cases had disparate clinical manifestations: one with mild disease which was identified in an outpatient setting, while a second case required intensive care unit-level support. Heartland virus genome sequences from the two cases were more similar to viruses from other states than they were to one another. InterpretationDespite only two prior reported cases of Heartland virus disease in North Carolina, we identified two individuals with acute HRTV viremia. Further surveillance for HRTV disease is necessary to understand the burden of disease and to facilitate further studies of virus pathogenesis and host responses. FundingFunding for the study was provided by a Creativity Hub Award to Dr. Boyce from the UNC Office of the Vice Chancellor for Research. Dr. Zychowskis effort was supported by the T32 NIAD grant AI070114.