Wastewater-based surveillance for influenza and respiratory syncytial virus: Insights from a 21-month study in Oklahoma.

Community-acquired respiratory viruses

Respiratory syncytial virus (RSV) is a major cause of serious acute lower respiratory tract infection in infants and the elderly. The lack of a licensed RSV vaccine calls for the development of vaccines with other targets and vaccination strategies. Here, we construct a recombinant protein, designated P-KFD1, comprising RSV phosphoprotein (P) and the E.-coli-K12-strain-derived flagellin variant KFD1. Intranasal immunization with P-KFD1 inhibits RSV replication in the upper and lower respiratory tract and protects mice against lung disease without vaccine-enhanced disease (VED). The P-specific CD4+ Tcells provoked by P-KFD1 intranasal (i.n.) immunization either reside in or migrate to the respiratory tract and mediate protection against RSV infection. Single-cell RNA sequencing (scRNA-seq) and carboxyfluorescein succinimidyl ester (CFSE)-labeled cell transfer further characterize the Th1 and Th17 responses induced by P-KFD1. Finally, we find that anti-viral protection depends on either interferon-γ (IFN-γ) or interleukin-17A (IL-17A). Collectively, P-KFD1 is a promising safe and effective mucosal vaccine candidate for the prevention of RSV infection.

Infections, Immunity & their Effects on Asthma

Background Respiratory viral infection (RVI) is a significant complication in patients with hematologic malignancies (HM). While risk factors and outcomes of severe infections have been studied in allogeneic hematopoietic cell transplant (HCT) recipients, data is limited for patients with lymphoma and multiple myeloma (MM). We investigated the risk factors and outcomes associated with severe respiratory syncytial virus (RSV) or influenza virus (IFV) infections from a large cohort of patients with lymphoma or MM.Figure 1:Respiratory viral infections per quarter stratified by viral pathogen.The dashed line represents the beginning of the COVID-19 era in the USA (March 2020).Abbreviations: IFV=influenza; Q=quarter; RSV= respiratory syncytial virus. Methods We performed a retrospective study in adult patients with lymphoma or MM who were diagnosed with RSV or IFV RVIs between 2016 and 2022 and followed up to 1 year. Primary outcomes were progression to lower respiratory tract infection (LRI) and all-cause 30- and 90-day mortality.Figure 2:Respiratory viral infections per quarter stratified by site of infection.The dashed line represents the beginning of the COVID-19 era in the USA (March 2020).The LRI group includes patients who presented with LRI or progressed from URI to LRI.Abbreviations: LRI=lower respiratory tract infection; Q=quarter; URI= upper respiratory tract infection Results We analyzed 440 patients with 490 consecutive viral episodes: 297 (61%) in patients with MM, and 193 (39%) with lymphoma. RVIs were secondary to RSV and IFV in 258 (52%) and 234 (48%) episodes, respectively. A decline in RVIs since the onset of the COVID-19 pandemic was noted, with an uptrend in the last quarter of 2022 (Figures 1, 2). Upon presentation, 303 (62%) patients were diagnosed with upper respiratory tract infection (URI) and 187 (38%) with LRI; while 19 patients (6%) with URI progressed to LRI. During follow-up, 57% were hospitalized, 8% were admitted to the ICU, 20 (4%) died within 30 days, and 32 (7%) within 90 days (Table 1). On multivariable analysis, RSV infection (vs. IFV), current/former smoking, recent steroid exposure, and lymphopenia or renal injury, were associated with LRI. Survival analysis revealed an association between MM (vs. lymphoma), current/former smoking, lymphopenia, and nosocomial infection and increased 30-day mortality risk, whereas LRI (vs. URI), current/former smoking, and lymphopenia were associated with 90-day mortality (Table 2).Table 1:Baseline characteristics and clinical outcomes following respiratory viral infections by site of infection.Abbreviations: BiPAP= bilevel positive airway pressure; CAR-T= chimeric antigen receptor T-cell therapy; COVID-19=Coronavirus Disease 2019; HCT= hematopoietic stem cell transplantation; HFNC= high-flow nasal cannula; HM= hematologic malignancy; ICU=intensive care unit; IQR= interquartile range; IVIG= Intravenous immunoglobulin; LRI= lower respiratory tract infection; RSV= respiratory syncytial virus; RVI= respiratory virus infection; SD=standard deviation; URI= upper respiratory tract infection. Conclusion We identified risk factors in lymphoma and MM patients with RVIs associated with substantial morbidity and mortality. Clinical parameters such as smoking status, recent steroid exposure, lymphopenia, and renal injury could potentially identify high-risk patients, thereby enabling better management strategies.Table 2:Independent predictors of LRI, 30-day and 90-day mortality by multivariable logistic regression.Abbreviations: aOR=Adjusted-Odds Ratio; LRI= lower respiratory tract infection; RSV= respiratory syncytial virus; RVI=respiratory tract infection; URI= upper respiratory tract infection; 95% CI= 95% Confidence Interval. Disclosures Fareed Khawaja, MBBS, Eurofins Viracor: Grant/Research Support|Symbio: Grant/Research Support Roy F. Chemaly, MD/MPH, AiCuris: Advisor/Consultant|AiCuris: Grant/Research Support|Ansun Pharmaceuticals: Advisor/Consultant|Ansun Pharmaceuticals: Grant/Research Support|Astellas: Advisor/Consultant|Eurofins-Viracor: Grant/Research Support|InflaRX: Advisor/Consultant|Janssen: Advisor/Consultant|Karius: Advisor/Consultant|Karius: Grant/Research Support|Merck/MSD: Advisor/Consultant|Merck/MSD: Grant/Research Support|Moderna: Advisor/Consultant|Oxford Immunotec: Advisor/Consultant|Oxford Immunotec: Grant/Research Support|Roche/Genentech: Advisor/Consultant|Roche/Genentech: Grant/Research Support|Shinogi: Advisor/Consultant|Takeda: Advisor/Consultant|Takeda: Grant/Research Support|Tether: Advisor/Consultant

Coronaviruses OC43, 229E, NL63, and HKU1 are endemic human respiratory coronaviruses that typically cause mild to moderate upper respiratory infections, similar to the common cold. They also may cause simple and complicated lower respiratory infections, otitis media, asthma exacerbations, gastroenteritis, and a few systemic complications. These viruses are usually seasonal (with winter dominance) and affect nearly all age groups. The seasonal and annual variation in virus prevalence has implications for understanding the concept of acquired immunity and its persistence or diminution. Coronaviruses generally have outbreak potential in susceptible populations of any age, particularly in patients with comorbidities, who tend to have increased clinical disease. These 4 coronaviruses are often found in the context of what appears to be coinfection with other pathogens, but especially other viruses. If coronaviruses are not specifically tested for, the sole detection of a viral copathogen would suggest the pathogen is the causative agent, when a coronavirus may be culpable, or both. The detection of these viruses in circumstances where respiratory viruses are generally sought in clinical samples is, therefore, justified. These pathogens can be chronically shed from the respiratory tract, which is more likely to occur among immunocompromised and complicated patients. These viruses share the potential for genetic drift. The genome is among the largest of RNA viruses, and the capability of these viruses to further change is likely underestimated. Given the potential disease among humans, it is justified to search for effective antiviral chemotherapy for these viruses and to consider uses in niche situations should effective therapy be defined. Whereas SARS-CoV-2 may follow the epidemiological pattern of SARS-CoV and extinguish slowly over time, there is yet concern that SARS-CoV-2 may establish itself as an endemic human respiratory coronavirus similar to OC43, 2299E, NL63, and HKU1. Until sufficient data are acquired to better understand the potential of SARS-CoV-2, continued work on antiviral therapy and vaccination is imperative.

Role of respiratory pathogens in infants hospitalized for a first episode of wheezing and their impact on recurrences

Inhibition of Respiratory Syncytial Virus Infections With Morpholino Oligomers in Cell Cultures and in Mice

Background Humoral immune responses to respiratory syncytial virus (RSV) infection in hematopoietic cell transplant (HCT) recipients have not been well characterized. We measured RSV pre-fusion F antibody (RSVpreF) titers in HCT recipients in 3 groups: upper respiratory tract infections (URTI) only, lower respiratory tract infections (LRTI) only, and those who progressed from URTI to LRTIs (progressors). Methods We evaluated adult and pediatric allogeneic HCT recipients who acquired RSV from 12/2011 to 12/2019. Prospective and leftover clinical serum specimens were collected from the Fred Hutchinson Cancer Center. Samples included baseline prior to infection, at RSV URTI or LRTI diagnosis, and after infection (both 4-6 and >8 weeks). Quantitative RSVpreF titers were determined using the MesoScale Discovery electrochemiluminescence assay. We compared longitudinal log10 RSVpreF titers among groups using linear mixed effect models with restricted cubic splines. Results A total of 129 samples from 39 patients were analyzed; 24 with URTI only, 8 progressors, and 7 with LRTI only. The median age at infection was 45 years, but HCT recipients with LRTI only had an overall higher median age of 54 (range 23-63) (Table 1). Most had acute leukemia (n=18, 46%) and received peripheral blood stem cells (n=27, 69%) as their cell source (Table 1). Median RSVpreF values were similar across the 3 different groups at each timepoint (Table 2). Trajectories of RSVpreF titers over time for each patient trended towards an increase following RSV diagnosis in all groups (Figure 1a). Predicted curves in RSVpreF titers over time did not differ significantly by group (Figure 1b, p=0.38).Table 2.Summary of RSVpreF titers among allogeneic transplant recipients before and after a post-HCT RSV infection. RSVpreF titers were obtained utilizing Meso Scale Discovery chemiluminescence assays. (A) RSVpreF titers for allogeneic HCT recipients with RSV infection. Each line represents a patient. The top panel shows patients with RSV URTI only, the middle panel shows patients who progressed from URTI to LRTI, and the bottom panel shows patients who presented with RSV LRTI. Open circles represent time points prior to RSV diagnosis, filled circles represent time points just prior to, on or after URTI, and asterisks represent time points just prior to, on, or after LRTI. (B) RSVpreF titers plotted by days from RSV diagnosis. Open circles represent observed values and lines represent model predicted trajectories. Black circles and lines correspond to patients with RSV URTI only, blue corresponds to patients who progressed from RSV URTI to LRTI, and dark orange represents patient who presented with RSV LRTI at diagnosis. An outlying observation at day -53 was excluded. Conclusion Among allogeneic HCT recipients with RSV infection, RSVpreF titers increased in response to RSV infection, but did not differ among patients with URTI only, progressors, or those with LRTI only across pre and post infection time points. Our novel immunoassay results are similar to those from previously described RSV neutralization assays, but extended the observation period beyond 4 weeks after infection, revealing a modest increase in RSVpreF titers. Further characterization of RSV humoral immunity, including mucosal immunity, is needed. Disclosures chikara Ogimi, MD, bioMerieux Japan Ltd.: Honoraria|Horiba: Honoraria|Pfizer: Honoraria Janet A. Englund, MD, Ark Biopharma: Advisor/Consultant|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Meissa Vaccines: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Moderna: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant Emily T. Martin, PhD, MPH, Merck: Grant/Research Support Michael J. Boeckh, MD PhD, Allovir: Advisor/Consultant|Amazon: Grant/Research Support|Ansun: Grant/Research Support|Merck: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Symbio: Advisor/Consultant Alpana Waghmare, MD, Allovir: Grant/Research Support|Amazon: Grant/Research Support|Ansun Biopharma: Grant/Research Support|GlaxoKlineSmith/Vir: Grant/Research Support|Kyorin Pharmaceuticals: Advisor/Consultant|Pfizer: Grant/Research Support|Vir Biotechnology: Advisor/Consultant

Background Wastewater-based surveillance (WBS) of respiratory syncytial virus (RSV) detected seasonal activity up to one month earlier than clinical surveillance (CS) during the 2022 RSV season in two Ontario cities, Canada. The associated cost-consequence analysis comparing the use of WBS vs CS to guide the start of provincial immunoprophylaxis programs showed WBS-guided prophylaxis realized savings of CAD $2.1-3.5m in the first year and $13.7-16.6m over 1-3 years due to early RSV detection. Further investigation is needed to explore the reproducibility and scalability of WBS for RSV in a provincial surveillance initiative. Geospatial distribution of the 12 sentinel sampling locations across Ontario, Canada Methods In collaboration with the Ontario Ministry of Health (MOH), 12 locations in the province were selected as sentinel WBS sites based on population density, socio-economic strata, and their significance as primary pediatric centres where palivizumab is administered (Figure 1). City-wide, 24-hour composite wastewater samples were collected and screened for RSV by RT-qPCR three to seven times a week. We monitored RSV-WBS data and pediatric RSV-related hospitalizations (RSVH) between September 1, 2023, to March 15, 2024. Cumulative fraction of locations for the onset of the 2023 RSV season in Ontario, Canada Date of onset of regional RSV activity as identify by wastewater surveillance at each of the 12 sentinel locations during the 2023 RSV season, represented as a cumulative fraction. Results During the study period, RSV was detected in 76% (1092/1440) of all wastewater samples collected, with detection at all locations. WBS effectively tracked the geographical variation in viral activity across the province. There was a 73-day variation in the RSV season onset across locations, with northern sites lagging southern locations by 62 ± 11 days (Figure 2). WBS detected regional RSV activity up to 15 days prior to the provincial start date of October 30, 2023, as determined by CS. Considering the estimated WBS cost of $0.50 per eligible infant compared to an average of $5,000 per RSVH, and an estimated fewer RSVHs per year, provincial WBS-guided prophylaxis could significantly reduce healthcare costs and the burden of RSV associated illness. Conclusion RSV WBS could be a highly cost-effective strategy that identifies the early onset of the RSV season, thereby optimizing immunoprophylaxis timing. Additionally, it is scalable to communities of varying sizes and socioeconomic strata and reflects geospatial variation in regional RSV activity supporting its use in a provincial-wide WBS initiative. Disclosures Bosco Paes, MD, Professor Emeritus, AstraZeneca: advisor/lecturer outside the scope of this study|Sanofi: advisor/lecturer outside the scope of this study John Fullarton, n/a, AstraZeneca: Employer has received payment from AstraZeneca for work on various projects outside the scope of this study|Sanofi: Employer has received payment from Sanofi for work on various projects outside the scope of this study Barry Rodgers-Gray, n/a, AstraZeneca: Employer has received payment from AstraZeneca for work on various projects outside the scope of this study|Sanofi: Employer has received payment from Sanofi for work on various projects outside the scope of this study

The aim of this study was to evaluate whether community-level monitoring of respiratory and enteric viruses in wastewater can provide a comprehensive picture of local virus circulation. Wastewater samples were collected weekly at the wastewater treatment plant (WWTP) inlet and at the outlet of a nearby nursing home (NH) in Burgundy, France, during the winter period of 2022/2023. We searched for the pepper mild mottle virus as an indicator of fecal content as well as for the main respiratory viruses [severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza, and respiratory syncytial virus] and enteric viruses (rotavirus, sapovirus, norovirus, astrovirus, and adenovirus). Samples were analyzed using real-time reverse transcription PCR-based methods. SARS-CoV-2 was the most frequently detected respiratory virus, with 66.7% of positive samples from the WWTP and 28.6% from the NH. Peaks of SARS-CoV-2 were consistent with the chronological incidence of infections recorded in the sentinel surveillance and the nearby hospital databases. The number of positive samples was lower in the NH than in WWTP for the three respiratory viruses. Enteric viruses were frequently detected, most often sapovirus and norovirus genogroup II, accounting both for 77.8% of positive samples in the WWTP and 57.1% and 37%, respectively, in the NH. The large circulation of sapovirus was unexpected in particular in the NH. Combined wastewater surveillance using simple optimized methods can be a valuable tool for monitoring viral circulation and may serve as a suitable early warning system for identifying both local outbreaks and the onset of epidemics. These results encourage the application of wastewater-based surveillance (WBS) to SARS-CoV2, norovirus, and sapovirus.IMPORTANCEWBS provides valuable information on the spread of epidemic viruses in the environment using appropriate and sensitive detection methods. By monitoring the circulation of viruses using reverse transcription PCR methods in wastewater from the inlet of a wastewater treatment plant and the outlet of a nearby retirement home (connected to the same collective sewer network), we aimed to demonstrate that implementing combined WBS at key community sites allows effective detection of the occurrence of respiratory (influenza, respiratory syncytial virus, and SARS-CoV-2) and enteric (norovirus, rotavirus, and sapovirus) virus infections within a given population. This analysis on a localized scale provided new information on the viral circulation in the two different sites. Implementing WBS to monitor the circulation or the emergence of infectious diseases is an important means of alerting the authorities and improving public health management. WBS could participate actively to the health of humans, animals, and the environment.

Comparison of Epidemiologic and Clinical Features between Respiratory Syncytial Virus and Human Metapneumovirus in Korean Children

Respiratory syncytial virus (RSV) remains one of the most important infectious causes of hospitalization in infants and children. This enveloped, RNA virus produces predictable yearly outbreaks of disease that typically peak between January and February in countries in the northern hemisphere.1 The outcome of RSV infection varies from mild upper respiratory tract infection in approximately 75% of infected infants and young children to severe life-threatening disease in a small percent of infected patients.2 In the United States, RSV lower respiratory tract infection accounts for nearly 50% of hospitalizations due to bronchiolitis and 25% of hospitalizations due to pneumonia.1 Serologic surveys suggest that by 2 years of age, more than 90% of all children have been infected by RSV.3 Whether RSV infection early in life predisposes to subsequent reactive airway disease remains an unanswered question. Reinfection is common, indicating that immunity to RSV following natural infection is less than complete.4-6 RSV lower respiratory tract disease occurs primarily in infants under 2 years of age; most infants who require hospitalization are previously healthy infants less than 6 months of age. Premature infants, infants born with congenital heart disease, and those with chronic lung disease (such as bronchopulmonary dysplasia [BPD]) constitute additional high-risk groups with high rates of hospitalization due to RSV infection.7-13 A recent report describes RSV mortality rates among such hospitalized infants of 4% to 5%.14Pre-engraftment bone marrow transplant recipients, solid organ transplant recipients, and lymphopenic children receiving chemotherapy appear to suffer even higher mortality rates, although prospective data are not available.15 Morbidity in these groups is also great; the average hospital stay and intensity of care for such children may be several times that of previously healthy infants. Despite the importance of RSV as a pathogen in the pediatric age group, …

During a 2-year period, 157 consecutive episodes of respiratory virus infections that occurred in 130 patients with upper or lower respiratory tract infection were analyzed for respiratory viruses. A respiratory virus was identified in 75 episodes (48%), and several viruses were found in 13 episodes: there were a total of 56 influenza A virus infections, 14 respiratory syncytial virus infections, 8 adenovirus infections, 8 infections with parainfluenza virus types 1 or 3, and 7 enterovirus infections. On multivariate analysis, the only variable that predicted progression to pneumonia in patients with an upper respiratory tract infection was the presence of respiratory syncytial virus, whereas lymphocytopenia had a nonsignificant trend. Also, among the 38 patients who had pneumonia at any time during the episode, both respiratory syncytial virus and lymphocytopenia were commonly found. For both epidemiological and therapeutic considerations, frequent screening for respiratory viruses should be incorporated into the routine diagnostic study of patients with hematologic malignancies.

Nuclear-localized human respiratory syncytial virus NS1 protein modulates host gene transcription.

Alterations in Pulmonary Function Following Respiratory Viral Infection