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Proceedings of the National Academy of... Oct 2023As SARS-CoV-2 variants of concern (VoCs) that evade immunity continue to emerge, next-generation adaptable COVID-19 vaccines which protect the respiratory tract and...
As SARS-CoV-2 variants of concern (VoCs) that evade immunity continue to emerge, next-generation adaptable COVID-19 vaccines which protect the respiratory tract and provide broader, more effective, and durable protection are urgently needed. Here, we have developed one such approach, a highly efficacious, intranasally delivered, trivalent measles-mumps-SARS-CoV-2 spike (S) protein (MMS) vaccine candidate that induces robust systemic and mucosal immunity with broad protection. This vaccine candidate is based on three components of the MMR vaccine, a measles virus Edmonston and the two mumps virus strains [Jeryl Lynn 1 (JL1) and JL2] that are known to provide safe, effective, and long-lasting protective immunity. The six proline-stabilized prefusion S protein (preS-6P) genes for ancestral SARS-CoV-2 WA1 and two important SARS-CoV-2 VoCs (Delta and Omicron BA.1) were each inserted into one of these three viruses which were then combined into a trivalent "MMS" candidate vaccine. Intranasal immunization of MMS in IFNAR1 mice induced a strong SARS-CoV-2-specific serum IgG response, cross-variant neutralizing antibodies, mucosal IgA, and systemic and tissue-resident T cells. Immunization of golden Syrian hamsters with MMS vaccine induced similarly high levels of antibodies that efficiently neutralized SARS-CoV-2 VoCs and provided broad and complete protection against challenge with any of these VoCs. This MMS vaccine is an efficacious, broadly protective next-generation COVID-19 vaccine candidate, which is readily adaptable to new variants, built on a platform with a 50-y safety record that also protects against measles and mumps.
Topics: Cricetinae; Animals; Humans; Mice; SARS-CoV-2; COVID-19 Vaccines; Mumps; COVID-19; Measles-Mumps-Rubella Vaccine; Antibodies, Viral; Broadly Neutralizing Antibodies; Immunoglobulin G; Measles; Mesocricetus; Antibodies, Neutralizing; Spike Glycoprotein, Coronavirus
PubMed: 37796985
DOI: 10.1073/pnas.2220403120 -
Journal of Clinical Virology : the... Aug 2024Measles, mumps, and rubella(MMR) vaccination is critical to measles outbreak responses. However, vaccine reactions and detection of measles vaccine RNA in recently...
BACKGROUND
Measles, mumps, and rubella(MMR) vaccination is critical to measles outbreak responses. However, vaccine reactions and detection of measles vaccine RNA in recently immunized persons may complicate case classification especially in those presenting with another respiratory viral illness. We aim to characterize cases of measles vaccine shedding in recently vaccinated children presenting with respiratory viral symptoms.
METHODS
Children who were tested with a multiplex respiratory panel <30 days after receiving MMR were identified. Remnant nasopharyngeal(NP) samples were tested for measles vaccine by PCR. Medical records were reviewed for demographics, presenting symptoms, and test results.
RESULTS
From January 2022 to March 2023, 127 NP from children who received MMR were tested. Ninety-six NP were collected after the first dose, of which 33(34.4 %) were positive for vaccine RNA. The median interval between MMR and detection was 11.0 days. Thirty-one NP were collected after the second MMR and 1(3.2 %) was positive; time between the vaccination and detection was 18.9 days. Median cycle threshold(Ct) value of the measles PCR for vaccine shedding was significantly higher than median Ct in children with wild-type infection.
CONCLUSION
Shedding of measles vaccine RNA is not uncommon and vaccine RNA can be detected up to 29 days post MMR; the amount of vaccine RNA shedding is low indicated by high Ct values. Clinicians and public health officials should consider performing measles vaccine testing on those testing positive for measles within one month of MMR vaccination, especially if the Ct value is high and definitive epidemiological links are absent.
Topics: Humans; Measles-Mumps-Rubella Vaccine; Female; Male; Virus Shedding; RNA, Viral; Child, Preschool; Infant; Vaccination; Child; Measles; Nasopharynx; Mumps; Rubella; Adolescent
PubMed: 38823291
DOI: 10.1016/j.jcv.2024.105696 -
Autophagy May 2024Macroautophagy/autophagy is a tightly regulated cellular process integral to homeostasis and innate immunity. As such, dysregulation of autophagy is associated with...
Macroautophagy/autophagy is a tightly regulated cellular process integral to homeostasis and innate immunity. As such, dysregulation of autophagy is associated with cancer, neurodegenerative disorders, and infectious diseases. While numerous factors that promote autophagy have been characterized, the key mechanisms that prevent excessive autophagy are less well understood. Here, we identify CSNK2/CK2 (casein kinase 2) as a negative regulator of autophagy. Pharmacological inhibition of CSNK2 activity or siRNA-mediated depletion of CSNK2 increased basal autophagic flux in cell lines and primary human lung cells. , ectopic expression of CSNK2 reduced autophagic flux. Mechanistically, CSNK2 interacted with the FLN (filamin)-NHL domain-containing tripartite motif (TRIM) family members TRIM2, TRIM3 and TRIM71. Our data show that recruitment of CSNK2 to the C-terminal NHL domain of TRIM3 lead to its robust phosphorylation at serine 661 by CSNK2. A phosphorylation-defective mutant of TRIM3 was unable to reduce autophagosome numbers indicating that phosphorylation by CSNK2 is required for TRIM-mediated autophagy inhibition. All three TRIMs facilitated inactivation of the ULK1-BECN1 autophagy initiation complex by facilitating ULK1 serine 757 phosphorylation. Inhibition of CSNK2 promoted autophagy upon influenza A virus (IAV) and measles virus (MeV) infection. In line with this, targeting of CSNK2 or depletion of TRIM2, TRIM3 or TRIM71 enhanced autophagy-dependent restriction of IAV, MeV and human immunodeficiency virus 1 (HIV-1). Thus, our results identify the CSNK2-TRIM2, -TRIM3, -TRIM71 axis as a key regulatory pathway that limits autophagy. Targeting this axis may allow for therapeutic induction of autophagy against viral infections and in diseases associated with dysregulated autophagy. ATG5: autophagy related 5; BafA1: bafilomycin A; BECN1: beclin 1; CCD: coiled-coil domain; CSNK2/CK2: casein kinase 2; CSNK2A1: casein kinase 2 alpha 1; CSNK2A2: casein kinase 2 alpha 2; CSNK2B: casein kinase 2 beta; FLN: filamin; HeLa GL: HeLa cells stably expressing eGFP-LC3B; HIV-1: human immunodeficiency virus 1; IAV: influenza A virus; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3; MeV: measles virus; MTOR: mechanistic target of rapamycin kinase; RING: really interesting new gene; SQSTM1/p62: sequestosome 1; TRIM: tripartite motif; ULK1: unc-51 like autophagy activating kinase 1.
Topics: Humans; Autophagy; Casein Kinase II; Tripartite Motif Proteins; Autophagy-Related Protein-1 Homolog; Phosphorylation; HEK293 Cells; Beclin-1; Intracellular Signaling Peptides and Proteins; Autophagosomes; HeLa Cells; Ubiquitin-Protein Ligases; Carrier Proteins
PubMed: 37938186
DOI: 10.1080/15548627.2023.2281128 -
Frontiers in Public Health 2023Despite the outstanding measles vaccine coverage (MVC) in Eritrea, sporadic outbreaks are not uncommon. Therefore, understanding the incidence of laboratory-confirmed...
BACKGROUND
Despite the outstanding measles vaccine coverage (MVC) in Eritrea, sporadic outbreaks are not uncommon. Therefore, understanding the incidence of laboratory-confirmed measles virus cases, related factors, and spatial inequalities in testing and surveillance remains crucial. In this analysis, we evaluated the incidence and spatiotemporal distribution of measles in Eritrea. An evaluation of the factors associated with measles vaccination and IgM positive (+) febrile rash was also undertaken.
METHODS
A retrospective (period: 2002-2020) study was carried out by abstracting data from the integrated disease surveillance and response database (IDSR). Data was analyzed using descriptive statistics and binary logistic regression. Spatial variability and distribution of confirmed cases was evaluated using ArcGIS Pro version 3.0.1.
RESULTS
In total, 9,111 suspected cases, 2,767 [1,431 (51.7%) females] were serologically tested. The median (IQR) age, minimum-maximum age were 7 years (IQR: 4-14 years) and 1 month-97 years, respectively. Among the 608(21.9%) laboratory-confirmed cases, 534 (87.8%) were unvaccinated and 53 (9.92%) were < 1 year old. The crude incidence rate for MV was 14/100,000 persons. The age-specific positivity rate per 100,000 suspected cases tested was 21.5 with individuals >30 years presenting with the highest rates (69.9/100,000). Higher odds (OR) of MV test positivity was associated with age at onset - higher in the following age-bands [10-14 years: OR = 1.6 (95%CI, 1.1-2.2, value of = 0.005); 15-29 years: OR = 7.0 (95%CI, 5.3-9.2, value of = 0.005); and > =30 years: OR = 16.7 (95%CI, 11.7-24) < 0.001]. Other associations included: Address - higher in Anseba (OR = 2.3, 95%CI: 1.7-3.1, value of <0.001); Debub (OR = 2.7, 95%CI: 1.9-3.9, value of < 0.001); Gash-Barka (OR = 15.4, 95%CI: 10.9-21.7, value of < 0.001); Northern Red Sea (OR = 11.8, 95%CI: 8.5-16.2, value of < 0.001); and Southern Red Sea (OR = 14.4, 95%CI: 8.2-25.2, value of < 0.001). Further, test positivity was higher in health centers (OR = 2.5, 95%CI: 1.9-3.4, value of < 0.001) and hospitals (OR = 6.8, 95%CI: 5.1-9.1, value of < 0.001). Additional factors included vaccination status - higher in the unvaccinated (OR = 14.7, 95%CI: 11.4-19.1, value of < 0.001); and year of onset of rash - (higher >2015: OR = 1.4, 95%CI: 1.1-1.7, value of <0.001). Uptake of measles vaccine associated with a similar complement of factors.
CONCLUSION
In large part, efforts to eliminate measles in Eritrea are hindered by disparities in vaccine coverage, under-surveillance, and low vaccination rates in neighboring countries. Enhanced surveillance and regional micro planning targeting hard-to-reach areas can be an effective strategy to improve measles elimination efforts in Eritrea.
Topics: Female; Humans; Infant; Adult; Child; Adolescent; Male; Measles virus; Incidence; Eritrea; Retrospective Studies; Measles; Measles Vaccine; Exanthema
PubMed: 37780421
DOI: 10.3389/fpubh.2023.1218317 -
JCI Insight Jul 2023We previously reported that measles virus nucleocapsid protein (MVNP) expression in osteoclasts (OCLs) of patients with Paget disease (PD) or targeted to the OCL lineage...
We previously reported that measles virus nucleocapsid protein (MVNP) expression in osteoclasts (OCLs) of patients with Paget disease (PD) or targeted to the OCL lineage in MVNP-transgenic mice (MVNP mice) increases IGF1 production in osteoclasts (OCL-IGF1) and leads to development of PD OCLs and pagetic bone lesions (PDLs). Conditional deletion of Igf1 in OCLs of MVNP mice fully blocked development of PDLs. In this study, we examined whether osteocytes (OCys), key regulators of normal bone remodeling, contribute to PD. OCys in PDLs of patients and of MVNP mice expressed less sclerostin, and had increased RANKL expression compared with OCys in bones from WT mice or normal patients. To test whether increased OCL-IGF1 is sufficient to induce PDLs and PD phenotypes, we generated TRAP-Igf1 (T-Igf1) transgenic mice to determine whether increased IGF1 expression in the absence of MVNP in OCLs is sufficient to induce PDLs and pagetic OCLs. We found that T-Igf1 mice at 16 months of age developed PD OCLs, PDLs, and OCys, with decreased sclerostin and increased RANKL, similar to MVNP mice. Thus, pagetic phenotypes could be induced by OCLs expressing increased IGF1. OCL-IGF1 in turn increased RANKL production in OCys to induce PD OCLs and PDLs.
Topics: Animals; Mice; Bone and Bones; Gene Expression; Mice, Transgenic; Osteitis Deformans; Osteoclasts; Osteocytes
PubMed: 37338990
DOI: 10.1172/jci.insight.159838 -
Nature Communications Jan 2024Measles virus (MV) vaccine strains have shown significant preclinical antitumor activity against glioblastoma (GBM), the most lethal glioma histology. In this first in...
Measles virus (MV) vaccine strains have shown significant preclinical antitumor activity against glioblastoma (GBM), the most lethal glioma histology. In this first in human trial (NCT00390299), a carcinoembryonic antigen-expressing oncolytic measles virus derivative (MV-CEA), was administered in recurrent GBM patients either at the resection cavity (Group A), or, intratumorally on day 1, followed by a second dose administered in the resection cavity after tumor resection on day 5 (Group B). A total of 22 patients received study treatment, 9 in Group A and 13 in Group B. Primary endpoint was safety and toxicity: treatment was well tolerated with no dose-limiting toxicity being observed up to the maximum feasible dose (2×10 TCID50). Median OS, a secondary endpoint, was 11.6 mo and one year survival was 45.5% comparing favorably with contemporary controls. Other secondary endpoints included assessment of viremia, MV replication and shedding, humoral and cellular immune response to the injected virus. A 22 interferon stimulated gene (ISG) diagonal linear discriminate analysis (DLDA) classification algorithm in a post-hoc analysis was found to be inversely (R = -0.6, p = 0.04) correlated with viral replication and tumor microenvironment remodeling including proinflammatory changes and CD8 + T cell infiltration in post treatment samples. This data supports that oncolytic MV derivatives warrant further clinical investigation and that an ISG-based DLDA algorithm can provide the basis for treatment personalization.
Topics: Humans; Oncolytic Viruses; Measles virus; Glioblastoma; Carcinoembryonic Antigen; Oncolytic Virotherapy; Neoplasm Recurrence, Local; Measles Vaccine; Tumor Microenvironment
PubMed: 38216554
DOI: 10.1038/s41467-023-43076-7 -
The Science of the Total Environment Sep 2023A multiplex quantitative reverse transcription polymerase chain reaction (RT-qPCR)-based method was designed for the simultaneous detection of influenza A, SARS-CoV-2,...
A multiplex quantitative reverse transcription polymerase chain reaction (RT-qPCR)-based method was designed for the simultaneous detection of influenza A, SARS-CoV-2, respiratory syncytial virus, and measles virus. The performance of the multiplex assay was compared to four monoplex assays for relative quantification using standard quantification curves. Results showed that the multiplex assay had comparable linearity and analytical sensitivity to the monoplex assays, and the quantification parameters of both assays demonstrated minimal differences. Viral reporting recommendations for the multiplex method were estimated based on the corresponding limit of quantification (LOQ) and the limit of detection at 95 % confidence interval (LOD) values for each viral target. The LOQ was determined by the lowest nominal RNA concentrations where %CV values were ≤35 %. Corresponding LOD values for each viral target were between 15 and 25 gene copies per reaction (GC/rxn), and LOQ values were within 10 to 15 GC/rxn. The detection performance of a new multiplex assay was validated in the field by collecting composite wastewater samples from a local treatment facility and passive samples from three sewer shed locations. Results indicated that the assay could accurately estimate viral loads from various sample types, with samples collected from passive samplers showing a greater range of detectable viral concentrations than composite wastewater samples. This suggests that the sensitivity of the multiplex method may be improved when paired with more sensitive sampling methods. Laboratory and field results demonstrate the robustness and sensitivity of the multiplex assay and its applicability to detect the relative abundance of four viral targets among wastewater samples. Conventional monoplex RT-qPCR assays are suitable for diagnosing viral infections. However, multiplex analysis using wastewater provides a fast and cost-effective way to monitor viral diseases in a population or environment.
Topics: Humans; Respiratory Syncytial Viruses; SARS-CoV-2; Influenza, Human; Wastewater; Sensitivity and Specificity; COVID-19; Virus Diseases; Measles; Multiplex Polymerase Chain Reaction
PubMed: 37201817
DOI: 10.1016/j.scitotenv.2023.164261 -
Euro Surveillance : Bulletin Europeen... Feb 2024We report on an ongoing measles outbreak in the Federation of Bosnia and Herzegovina with 141 cases notified between week 52 2023 and week 6 2024. Among those with known...
We report on an ongoing measles outbreak in the Federation of Bosnia and Herzegovina with 141 cases notified between week 52 2023 and week 6 2024. Among those with known vaccination status, 97% were unvaccinated and the most affected group is children under the age of 5 years (n = 87) who were not vaccinated during the pandemic years. Sixty-eight cases were hospitalised, the most common complications were measles-related pneumonia and diarrhoea. The sequenced measles viruses from four cases belonged to genotype D8.
Topics: Child; Humans; Child, Preschool; Vaccination; Bosnia and Herzegovina; Measles; Measles virus; Disease Outbreaks; Exanthema
PubMed: 38426241
DOI: 10.2807/1560-7917.ES.2024.29.9.2400107 -
Methods in Molecular Biology (Clifton,... 2024Morbilliviruses such as measles virus (MeV) are responsible for major morbidity and mortality worldwide, despite the availability of an effective vaccine and global...
Morbilliviruses such as measles virus (MeV) are responsible for major morbidity and mortality worldwide, despite the availability of an effective vaccine and global vaccination campaigns. MeV belongs to the mononegavirus order of viral pathogens that store their genetic information in non-segmented negative polarity RNA genomes. Genome replication and viral gene expression are carried out by a virus-encoded RNA-dependent RNA polymerase (RdRP) complex that has no immediate host cell analog. To better understand the organization and regulation of the viral RdRP and mechanistically characterize antiviral candidates, biochemical RdRP assays have been developed that employ purified recombinant polymerase complexes and synthetic RNA templates to monitor the initiation of RNA synthesis and RNA elongation in vitro. In this article, we will discuss strategies for the efficient expression and preparation of mononegavirus polymerase complexes, provide detailed protocols for the execution and optimization of RdRP assays, evaluate alternative options for the choice of template and detection system, and describe the application of the assay for the characterization of inhibitor candidates. Although MeV RdRP assays are the focus of this article, the general strategies and experimental approaches are readily transferable to related viruses in the mononegavirus order.
Topics: Measles virus; RNA-Dependent RNA Polymerase; Virus Replication; RNA, Viral; Mononegavirales; Animals; Viral Proteins; Humans
PubMed: 38743360
DOI: 10.1007/978-1-0716-3870-5_3