Viruses-Basel最新文献

Chimeric orthoflaviviruses derived from the insect-specific Binjari virus (BinJV) offer a promising basis for safe orthoflavivirus vaccines. However, these vaccines have so far only been produced using adherent C6/36 Aedes albopictus mosquito cell cultures grown in serum-supplemented media, limiting their scalable manufacture. To address this, we adapted C6/36 cells for serum-free suspension culture using Sf900-III medium, achieving high peak cell densities (up to 2.5 × 10⁷ cells/mL). Higher agitation rates reduced cell aggregation, and cryopreservation and direct-to-suspension revival were successful, confirming the adapted line's stability for research and industrial applications. Despite this, BinJV-based chimeric orthoflaviviruses, including BinJV/WNV_KUN, a candidate vaccine for West Nile virus, and similar vaccines (BinJV/DENV2 and BinJV/JEV_NSW22) for dengue 2 virus and Japanese encephalitis virus, respectively, exhibited substantially reduced titres in C6/36 cultures infected in Sf900-III, a phenomenon attributed to the medium's acidic pH. Switching to the more alkaline, serum-free CD-FortiCHO medium enhanced the replication of these chimeric viruses to peak titres between 1.7 × 10⁷ and 7.6 × 10⁹ infectious units per mL whilst preserving viral integrity. These findings suggest that suspension-adapted C6/36 cultures in CD-FortiCHO medium can support high-yield vaccine production for various orthoflaviviruses and highlight the important role of cell culture media pH for orthoflavivirus bioprocessing. This scalable mosquito cell-based system could reduce production costs and improve vaccine accessibility, supporting efforts to combat arbovirus-related public health challenges.

Enterovirus D68 (EV-D68) is a leading cause of acute respiratory disease outbreaks, especially among children. EV-D68 infections can rapidly progress to severe clinical complications and potentially fatal outcomes. In Brazil, no diagnostic or genomic surveillance of this virus is currently performed. Between July and September 2023, cases of acute EV-D68 infection were identified among pediatric patients in several municipalities within the State of Alagoas, Northeast Brazil. Infections were confirmed by RT-qPCR using nasopharyngeal samples, and the complete EV-D68 genomes were sequenced and analyzed through phylogenetic inference. EV-D68 RNA was identified in four children aged 1-9 years from four geographically distinct municipalities in Alagoas. All infections were associated with lower respiratory tract symptoms, including dyspnea and wheezing; however, no fatalities were reported. Complete genomic sequencing revealed that the samples belonged to genotype B, subgenotype B3. This is the first study to report complete genomic data on EV-D68 infections from Brazil and South America. Enhanced genomic surveillance and focused EV-D68 diagnosis are critical to better understanding and managing the regional and national dissemination of this virus.

People with HIV (PWH) receiving antiretroviral therapy (ART), despite a similar life expectancy, have a higher incidence of comorbidities than the general population. This study assessed the influence of proinflammatory biomarkers and clinical factors on mortality of PWH. We included PWH hospitalized from 2009 to 2014 who continued ART until 2023. The baseline lipid profile, CD4+ cell count, platelets, CRP, PCT, TNF-α, VCAM-1, and HCV and HBV coinfection were evaluated. Multivariable logistic regression was used to evaluate factors associated with mortality. Among 72 PWH, 19 were lost to a follow-up and 13 died before 2023. The mean follow-up was 12.07 years, while the mean time to death was 4.32 years. The main causes of death were cancer (n = 7) and drug-related death (n = 4). In the multivariate analysis, HCV coinfection, CRP ≥ 5 mg/L, PCT ≥ 0.05 ng/mL, and VCAM-1 ≥ 922 ng/mL were associated with higher odds of death. Although people who died had lower total cholesterol and triglyceride concentrations, these parameters were not associated with mortality. Determining HCV coinfections and CRP, PCT, and VCAM-1 levels may help identify PWH at increased risk of death for intensified monitoring. Care should also be taken of PWH with normal lipid parameters.

Contemporary ART as Dolutegravir (DTG) has significantly advanced antiretroviral therapy, but relatively few data are available on its impact on the emergence of HIV-1 drug resistance mutations (DRMs). Monitoring the emergence of INSTI-associated DRMs following the introduction of DTG in Suriname will provide general insight and guide national HIV treatment strategies. All people living with HIV (PLHIV) in Suriname, for whom an INSTI drug resistance test was requested between September 2019 and February 2024 (n = 20), were included. HIV-1 integrase gene sequences were determined using Sanger sequencing. INSTI-associated mutations were identified using the Stanford HIV Drug Resistance Database program. The majority of the participants (66.7%) harbored HIV-1 subtype B, and 33.3% were B-recombinant forms. In addition to the INSTI wildtype, a strain was revealed carrying E157EQ and one person harbored a highly INSTI-resistant strain (E138K, G140S, Q148H and N155H). The emergence of a highly INSTI-resistant HIV-1 strain in Suriname, with unusual mutations for ART-experienced PLHIV exposed to DTG as the only INSTI, accentuates the need for continuous monitoring of the emergence of INSTI drug resistance mutations, not only to enable timely interventions and optimized treatment outcomes for PLHIV, but also to steer the decision making for ART protocols, especially for second generation INSTIs.

In this article, we present an assessment of the risk of the potential introduction and spread of highly pathogenic avian influenza (HPAI) in cows in Bulgaria. In the spring of 2024, we witnessed an unprecedented spread of the virus in dairy herds in the USA. This crossing of interspecies barriers by the virus creates a real danger of pandemic manifestations in humans. The continued spread of H5N1 clade 2.3.4.4b in dairy populations and other mammalian species and efficient animal-to-animal transmission increases the risk of infection and subsequent spread of the virus in human populations. According to registers, as of 1 November 2022, a total of 559,544 cattle were bred in Bulgaria. The total number of dairy cows decreased by 5.2% year-on-year to 197,996. Farms breeding dairy cows as of 1 November 2022 were 12,439, which is 22.1% less than the previous year. The production of cow's milk in 2022 amounted to 748,278 thousand liters. Traditionally, the largest share in the total yield of cow's milk is occupied by the south-central region with 25.9%, followed by the southeastern region with 18.5%. Due to potential risk factors such as the high concentration of dairy cows in high-risk areas for avian influenza A, the possibility of HPAI jumping the interspecies barrier and spreading in dairy herds in Bulgaria is very high. We therefore set out to assess the risk of virus penetration in these herds.

Background: COVID-19 can cause acute pulmonary hypertension (PH), worsening outcomes in critically ill elderly patients. Point-of-care ultrasound (POCUS), assessing right ventricular hemodynamics, predicts short-term outcomes. This study examines the long-term impact of acute PH on mortality in elderly COVID-19 patients.

Methods: This retrospective long-term study analyzed data from patients over 70 years old with severe COVID-19 pneumonia admitted to a mixed 25-bed, level 3 intensive care unit (ICU). POCUS focused on systolic pulmonary artery pressure (sPAP) at admission. Mortality was evaluated 1000 days post-admission.

Results: The study included 130 patients, comprising 30 long-term survivors and 100 non-survivors, with a cumulative long-term mortality rate of 77%. Non-survivors had significantly higher sPAP values (39.1 ± 12.8 vs. 30.4 ± 9.2, p = 0.04), which were associated with long-term mortality in survival analysis.

Conclusion: Acute pulmonary hypertension (PH), reflected by elevated systolic pulmonary artery pressure (sPAP), is strongly associated with long-term mortality in elderly critically ill COVID-19 patients. Early assessment of sPAP via POCUS may help identify high-risk patients and guide management strategies to improve outcomes.

High-risk human papillomavirus (HR-HPV) and other sexually transmitted infections (STIs-O) are promoters to the development of cervical cancer (CC), especially when they co-exist. This study aims to determine the prevalence of the major STIs-O and the rate of co-infection in women previously diagnosed with HR-HPV infection. For this observational study, 254 women aged 25-65 years who were being followed up for HR-HPV infection (without a CC history) were recruited at a hospital's Gynaecology Department from February 2024 to November 2024. Their endocervical specimens were collected and processed for HR-HPV, Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, and Trichomonas vaginalis detection by RT-PCR using commercially available reagents and equipment. The overall rate of infection was 38.6% for HPV and 4.3% for ITSs-O (3.8% in HPV-negative women and 5.1% in HPV-positive women). The presence of ITSs-O in women aged 25-34 was higher in those with a persistent positive result for HR-HPV (20.0% vs. 4.2%). Diverse multiple co-infections were found in HPV-positive women, whilst some single STIs-O were found in HPV-negative women. These results support the benefits of STI-O screening beyond an HR-HPV positive result, especially in those women under 35 years old.

Vaccination is a common influenza A virus (IAV) control strategy for pigs. Vaccine efficacy depends on strain cross-protection and effective vaccination program implementation. We evaluated a multi-faceted IAV vaccination strategy which included (a) monthly surveillance of pigs at weaning, (b) selection of epidemiologically relevant strains from farms under surveillance, (c) updating IAV strains in custom-made vaccines, and (d) seasonal mass vaccination with custom-made vaccines given to sows in 35 farrow-to-wean farms within an integrated swine farm system. Reduction of IAV in pigs from vaccinated sows was determined by monthly monitoring of farms for 30 months by IAV rRT-PCR (PCR) testing of nasal wipes collected from litters of piglets at weaning. Hemagglutinin (HA) nucleotide and amino acid (AA) sequence homology of the circulating and vaccine strains was determined by pairwise alignment and AA comparison at antigenic sites. Of the 35 farms monitored, 28 (80%) tested positive at least once, and 481 (5.75%) of 8352 PCR tests were IAV positive. Complete HA sequences were obtained from 54 H1 (22 H1-δ_1B.2.1, 28 H1-γ_1A.3.3.3, and 4 H1-pdm_1A.3.3.2 clades) and 14 H3 (12 IV-A 3.1990.4.1 and 2 IV-B 3.1990.4.2 clades) circulating IAV strains. During the study, custom-made vaccines were updated three times (eight strains total) and administered to sows at five distinct time periods. The HA AA similarity between vaccine and circulating strains ranged from 95% to 99%; however, the 0 to 71% similarity at HA antigenic sites prompted the vaccine updates. Herd IAV prevalence decreased from 40% (14/35) to 2.9% (1/35), accompanied by a numerical reduction in IAV-positive samples post-vaccination. Our results support having a comprehensive approach to controlling influenza in swine herds that includes surveillance, vaccination, and careful program implementation to reduce IAV in pigs.

The skin provides a life-sustaining interface between the body and the external environment. A dynamic communication among immune and non-immune cells in the skin is essential to ensure body homeostasis. Dysregulated cellular communication can lead to the manifestation of inflammatory skin conditions. In this review, we will focus on the following two key frontiers in the skin: innate immune sensors and cell death, as well as their cellular crosstalk in the context of skin homeostasis and inflammation. This review will highlight the recent advancements and mechanisms of how these pathways integrate signals and orchestrate skin immunity, focusing on inflammatory skin diseases and skin infections in mice and humans.

Gene therapy offers promising potential as an efficacious and long-lasting therapeutic option for genetic conditions, by correcting defective mutations using engineered vectors to deliver genetic material to host cells. Among these vectors, adeno-associated viruses (AAVs) stand out for their efficiency, versatility, and safety, making them one of the leading platforms in gene therapy. The enormous potential of AAVs has been demonstrated through their use in over 225 clinical trials and the FDA's approval of six AAV-based gene therapy products, positioning these vectors at the forefront of the field. This review highlights the evolution and current applications of AAVs in gene therapy, focusing on their clinical successes, ongoing developments, and the manufacturing processes required for the rapid commercial growth anticipated in the AAV therapy market. It also discusses the broader implications of these advancements for future therapeutic strategies targeting more complex and multi-systemic conditions and biological processes such as aging. Finally, we explore some of the major challenges currently confronting the field.