SD; standard deviation. [view World Health Organization. 6b. As previously observed by Perkmann et al. K18-hACE2 transgenic mice are highly susceptible and displayed clinical signs following SARS-CoV-2 challenge22,23. between patient and physician/doctor and the medical advice they may provide. Absorbance at 450nm was determined with a spectrophotometer. There are currently a few monoclonal antibody cocktails (such as bamlanivimab, casirivimab, and imdevimab together) that have been authorized by the US FDA for emergency use for the treatment of COVID-19 in certain population and similar medications have been authorized in other countries. Center of Excellence in Vaccine Research and Development (Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, Eakachai Prompetchara,Chutitorn Ketloy,Kittipan Tharakhet,Papatsara Kaewpang,Nongnaphat Yostrerat,Patrawadee Pitakpolrat,Supranee Buranapraditkun,Kanitha Patarakul,Teerasit Techawiwattanaboon,Tanapat Palaga&Kiat Ruxrungtham, Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, Eakachai Prompetchara,Chutitorn Ketloy,Kittipan Tharakhet&Patrawadee Pitakpolrat, Integrated Frontier Biotechnology for Emerging Disease, Chulalongkorn University, Bangkok, 10330, Thailand, Eakachai Prompetchara,Chutitorn Ketloy,Kanitha Patarakul&Kiat Ruxrungtham, Division of Infectious Diseases, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, 19104, USA, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, Thai Pediatric Gastroenterology, Hepatology and Immunology (TPGHAI) Research Unit, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand, Suwimon Manopwisedjaroen&Arunee Thitithanyanont, Virology and Cell Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, 12120, Thailand, Department of Virology, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, 10400, Thailand, Department of Veterinary Medicine, USAMD-AFRIMS, Bangkok, 10400, Thailand, BioNet-Asia, Co. Ltd, Bangkok, 10260, Thailand, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, Kanitha Patarakul&Teerasit Techawiwattanaboon, Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand, Genevant Sciences Corporation, Vancouver, BC, V5T 4T5, Canada, Department of Medicine, and School of Global Health, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand, You can also search for this author in Science 368, 489493 (2020). x2- p` ` \0`0e`X{StAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA^@%&D7(mylkv There is also a limitation regarding the two semi-quantitative antibody binding assays as a saturation limit could be reached because of their limited measurement range. Adv. Lv, J., Wu, H., Xu, J. Google Scholar. When compared with psVNT50 titers against BA.1, the GMT reduction against BA.4/5 in 10 and 30g dosed groups were 48 and 2.3 folds, respectively. For the Siemens assay, the optimal cutoff was within the same range as the reference cutoff (270 BAU/ml). Google Scholar. However, it has not been shown that COVID-19 mRNA vaccine encoding non-stabilized spike protein is not immunogenic or is not protective against viral challenge. Article wxWd~{Trru%m#97Z=}<8boK.3E@KT>1oW#!7q%7uJ?IC5 .iM!. These authors jointly supervised this work: Drew Weissman, Kiat Ruxrungtham. [ view less ], Affiliations: The authors would like to thanks Dr.Navapon Techakriengkrai, Faculty of Veterinary Science, Chulalongkorn University for providing HEK293T-hACE-2 cells. The structural study of S protein expressed by AZ1222 showed a native-like structure mostly found in the prefusion stage41. The mRNA vaccine technology transfer hub [updated 21 June 2022. The data as well as the p values suggested that the anti-S1 antibody reversed the impact of S1 on bursting activities. Developing highly effective vaccine platforms like mRNA technology in low- and middle-income countries (LMICs) is therefore an important goal21. In just over 2 years into the pandemic, more than 10 variants of the virus have been reported, of which, 5 variants, including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), and Omicron (B1.1.529) have been categorized by WHO as variants of concern (VOCs)7. PubMedGoogle Scholar. SARS-CoV-2 is the name of the virus that causes coronavirus disease 2019 (COVID-19). Walsh, E. E. et al. N Engl J Med 383, 24392450 (2020). Here, we describe the construction and preclinical evaluation of mRNA expressing the ectodomain of native, prefusion-non-stabilized S protein of wild-type (WT) Wuhan-Hu1 strain encapsulated within lipid nanoparticles, henceforth referred to as ChulaCov19. https://apps.who.int/iris/handle/10665/363344 (2022). *Important notice: bioRxiv publishes preliminary scientific reports that are not peer-reviewed and, therefore, should not be regarded as conclusive, guide clinical practice/health-related behavior, or treated as established information. This study also indicated that neutralizing S1 restores neuronal discharge activities to control levels. We suggest specific adjusted thresholds (BAU/ml) for the four commercial antibody assays that are used to assess pre-exposure prophylaxis in immunocompromised patients. Department of Infectious Diseases and Internal Medicine, Hpital Europen, Marseille, France, Affiliation: In contrast, undetectable fluorescent signals for S proteins were observed when HEK293T-hACE-2 were incubated with supernatant from untransfected cells (Fig. Source data are provided as a source data file. CoronaVac induces lower neutralising activity against variants of concern than natural infection. Moreover, the tissue slides were examined unblind. Protective activity of mRNA vaccines against ancestral and variant SARS-CoV-2 strains. The S protein trimer (S-trimer), depicted in Fig. COVID-19 treatments and pathogenesis including anosmia in K18-hACE2 mice. The team assessed the data using an algorithm devised in-house. Lancet Infect Dis 21, 13521354 (2021). tG gh3o^r3^Z\PQE5)?xLr3El79/iDm}y)MjbV|bBuD7l[U}\Cxo Comparative immunogenicity and reactogenicity of heterologous ChAdOx1-nCoV-19-priming and BNT162b2 or mRNA-1273-boosting with homologous COVID-19 vaccine regimens. N Engl J Med 383, 19201931 (2020). Please note that medical information found Vero E6 and HEK293T-hACE-2 were grown in Eagles minimum essential medium (EMEM) and Dulbeccos Modified Eagles Medium (DMEM), respectively supplemented with 5-10% heat-inactivated fetal bovine serum (HIFBS), 1% L-glutamine, 1% Pen/Strep, 40g/ml gentamicin and 0.25g/ml fungizone (all were from Invitrogen, Carlsbad, CA, USA) at 352 oC with 5% CO2. PN20-06). The new semi-quantitative testing service is the latest addition to the company's existing menu of COVID-19 qualitative IgG and IgG/IgM test services. The differences in the commercial assays used in this study are related to the components of the tests (the spike antigen epitopes used, the different isolates of the SARS-CoV-2, and the quantification of either total antibodies or only IgG) [2123]. These results reflect that ChulaCov19 was highly immunogenic and induced a Th1-skewed response in mice. Peletta, A. et al. ChulaCov19 significantly enhanced the magnitude of both NAb and T cell responses compared to homologous 2-dose regimens of either CoronaVac or AZD1222. . (2023, April 27). The results demonstrated that IgG2a/IgG1 (or Th1/Th2) ratios were greater than 1 in all vaccinated mice (Fig. KL and JH are employees of Genevant Sciences Corporation and are named on patent describing lipid nanoparticles. Broad and timely access to effective vaccines in LMICs, particularly the most under-served settings, has always been limited during past pandemics and this has extended to COVID-1920. The results should always be assessed in conjunction with patient's medical history, clinical presentation, and other findings. When considering a reference cutoff of 264 BAU/ml, the assays showed moderate to good agreement with Genscript sVNT, with strong variations of the kappa index from 0.52 for Beckman and Roche to 0.76 for Siemens (kappa = 0.72 for Abbott). b hACE-2 binding assay (merged): culture supernatant collected from ChulaCov19 transfected cells incubated with HEK293T- hACE-2 cells. It was subcloned into pUC-ccTEV-A101 using Afe I and Spe I restriction sites58. Today, hundreds of commercial antibody tests are on the market despite often lacking proper validation and with unsatisfactory sensitivity and/or specificity. Jiang, R. D. et al. The objective of the present study was to establish a new optimal threshold of protection for four different SARS-CoV-2 antibody assays [14]. Ff!T8$I$I$I In terms of spike-specific T-cell responses, our study found that AZD1222 prime/ChulaCov19 boost induced the highest magnitude of T cell response, superior to that of all tested regimens, including the homologous ChulaCov19 (Fig. Common SARS-CoV-2 virus antigenic targets include spike, envelope, and nucleocapsid proteins [1]. Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen. Pallesen, J. et al. PubMed Central p<0.05 and p<0.01 are indicated by * and **, respectively. 1a). Omicron stood out from other variants because it contained mutations that helped it evade immune cell protection. Alene, M. et al. Eichinger, K. M. et al. As expected, Omicron subvariants, especially BA.4/5, showed the largest drop in micro-VNT50 titers (Fig. Animals were immunized IM with 1g or 10g of ChulaCov19 at weeks 0 and 3. With such promising results from animal studies, the same formulation of ChulaCov19 vaccine that had been tested in animals is currently in phase 1-2 of clinical trials and can be manufactured locally for later clinical development. The data supporting the findings of this work are available within the paper and in the Supplementary Information file. The bands corresponding to S1, S2 and intact S (S0) were detected. In contrast, the optimal cutoff was higher for the Roche assay (559 BAU/ml). Additional quality control to ensure the absence of double-stranded RNA (dsRNA) and endotoxin contamination prior to encapsulation into lipid nanoparticles (LNPs) were performed as described previously60. on this website is designed to support, not to replace the relationship Previous study by Eichinger KM, et al. https://doi.org/10.1038/s41467-023-37795-0, DOI: https://doi.org/10.1038/s41467-023-37795-0. Goat-anti-human IgG, goat-anti-mouse IgG, or goat-anti-rabbit IgG antibodies (all were diluted 1:10,000) conjugated with horseradish peroxidase (HRP) were used as secondary antibodies (all were from KPL, MD, USA) and detected by chemiluminescence substrate (Immobilon western, Millipore, CA, USA) then exposed to an X-ray film. n=5 per group for Experiment 1, 2 and 3. b Challenge study in K18-hACE2 transgenic mice, n=6 in vaccinated groups and n=5 in control (PBS-receiving) group. The vaccine effectiveness is varied due to several factors such as the emergence of new variants, study population, and prevalence of the outbreak during the period the studies were conducted11,12,13. ADS Protection against WT (Wuhan-Hu1) viral challenge in K18-hACE2 transgenic mice mediated by ChulaCov19 was successfully demonstrated. Recombinant S protein with S1/S2 cleavage site abolished (ACROBioSystems, China) was used as positive control both in HEK293T-hACE-2 binding assay and western blot. 4a). In the present study, researchers quantified the neurological phenotypes induced in neurons by the SARS-CoV-2 S protein. A Single-Cycle Influenza A Virus-Based SARS-CoV-2 Vaccine Elicits Potent Immune Responses in a Mouse Model. PubMed Detection of antibodies to the SARS-CoV-2 spike glycoprotein in both serum and saliva enhances detection of infection All studies were conducted under protocols approved by the Committees on Care of Laboratory Animal Faculty of Medicine, Chulalongkorn University (IACUC approval no. Notably, SARS-CoV-2 RNA measured by ISH was undetected in lung tissues in mice vaccinated with ChulaCov19 at either 1 or 10 g dose. The titers were determined in duplicate assays from 5 mice in each group. Vaccines (Basel) 9, 874 (2021). World Health Organization (2022). c S protein expression in cell culture supernatant analyzed by western blot using anti-RBD, -S1, -S2 or PCS as primary antibody. CK was also funded by emerging Infectious Diseases and Vaccines Cluster, Ratchadapisek Sompoch Endowment Fund (2021), Chulalongkorn University (764002-HE04), and the Second Century Fund (C2F), Chulalongkorn University and Ratchadapiseksompotch Fund. Viruses were propagated in Vero E6 cells to generate sufficient titers 100TCID50 for the micro-VNT50 assay. 8 more], Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. 007/2563), and the Armed Forces Research Institute of Medical Sciences, AFRIMS (IACUC approval no. Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. . Secreted S protein was also subjected for analysis of its binding capability to hACE2. The most effective COVID-19 vaccines are mRNA-based and were first approved in the United Kingdom, the United States, and Europe. Nat Commun 13, 4710 (2022). Tseng, C. T. et al. "a97YEy111JlM7qqK;R]fr{g8 E]P7t iEx-m11tSmxsE,GE+hU#a=z1{/_vH}Nu&SENP_.*$ RL!DrojWs|[`}5C6nP,(n ,s-Km41vm8c/U3$@X3hUIwBge2Q{`4>4PQqo8"v3&v`wDXs%| 9>^8%|76sY6s$7PqI1QmO etbrr>$UmKd=UW-]Kd cg?q{`#*CM4\M6eKP2;:)U@(W$=u:{[9[S\2+wfynJ,%fd(~)qK5 Differences were considered significant at p<0.05 with exact p-values shown. Monoclonal anti-RBD (1:2,500), polyclonal-anti-S1 (1:5,000), -anti-S2 (1:5,000) or PSC (1:5,000) were used for detection of S protein in this step. Bars represent the meanSD of S-specific IFN- positive T cells after stimulated with overlapping peptide pools spanning the SARS-CoV-2 S1 (pooled #1-5) and S2 (pooled #6-10). Hunsawong, T. et al. In the homologous prime/boost of these 2 approved vaccines groups, each was given at four weeks interval. WW is an employee of BioNet-Asia, Co. Ltd. We have disclosed those interests fully to their affiliations, and we have in place an approved plan for managing any potential conflicts arising from licensing of the patents. In the same study, two doses of AZD1222 could protect rhesus macaque form viral challenge. The reaction was carried out employing T7 RNA polymerase (MegaScript, ThermoFisher Scientific, MA, USA) on a linearized plasmid (Not I/Afl II double digestions). This candidate vaccine has now completed non-clinical toxicity and biodistribution studies and has entered Phase 1 and 2 human trials. Klemis, V. et al. At week 5, mice were challenged intranasally with 2104 pfu of WT SARS-CoV-2. Stphane Blachier, Similar with the previous study, low level of viral RNA occasionally detected in survived mice was also reported by studies that used K18-hACE2 as a model28. Cevik, M. et al. 4b). Sci. Google Scholar. Few studies have highlighted the lack of standardization of SARS-CoV-2 serology, despite the use of the international standards set by the World Health Organization (WHO) for SARS-CoV-2 immunoglobulin levels (BAU/ml) [1013]. The absorbance was measured at a wavelength of 450nm using a Varioskan microplate reader (ThermoFisher Scientific, Vantaa, Finland). Nature Communications (Nat Commun) 4c). Six-day post challenge, wk5+6 days, mice were sacrificed to determine virus titers in different tissues (nasal turbinate, brain, lung, and kidney) and for histopathology. If testing will be delayed more than 7 days store at -20C or colder. Human codon-optimized sequences of the ectodomain of SARS-CoV-2 spike protein, amino acid position 1-1,210 (Wuhan Hu-1 complete genome, GenBank: MN908947.1, https://www.ncbi.nlm.nih.gov/nuccore/MN908947.1) was synthesized by GenScript, Piscataway, NJ, USA). Experiment 3: antibody durability and effect of 3rd dose of ChulaCov19 study, mice were immunized twice with 3 weeks interval with 5g of ChulaCov19 (1/10 of human dose used in clinical trial) then boosted again at week 20. An adjuvanted subunit SARS-CoV-2 spike protein vaccine provides protection against Covid-19 infection and transmission, Immunogenicity and protection of a variant nanoparticle vaccine that confers broad neutralization against SARS-CoV-2 variants, CpG-adjuvanted stable prefusion SARS-CoV-2 spike protein protected hamsters from SARS-CoV-2 challenge, Protection of hamsters challenged with SARS-CoV-2 after two doses of MVC-COV1901 vaccine followed by a single intranasal booster with nanoemulsion adjuvanted S-2P vaccine, mRNA-based SARS-CoV-2 vaccine candidate CVnCoV induces high levels of virus-neutralising antibodies and mediates protection in rodents, Intranasal immunization with a proteosome-adjuvanted SARS-CoV-2 spike protein-based vaccine is immunogenic and efficacious in mice and hamsters, Booster vaccination with Ad26.COV2.S or an Omicron-adapted vaccine in pre-immune hamsters protects against Omicron BA.2, The SARS-CoV-2 spike residues 616/644 and 1138/1169 delineate two antibody epitopes in COVID-19 mRNA COMIRNATY vaccine (Pfizer/BioNTech), A core-shell structured COVID-19 mRNA vaccine with favorable biodistribution pattern and promising immunity, https://www.ncbi.nlm.nih.gov/nuccore/MN908947.1, https://www.who.int/en/activities/tracking-SARS-CoV-2-variants, https://www.worldometers.info/coronavirus, https://covid19.trackvaccines.org/agency/who, https://apps.who.int/iris/handle/10665/363344, https://www.bloomberg.com/graphics/covid-vaccine-tracker-global-distribution, https://www.who.int/initiatives/the-mrna-vaccine-technology-transfer-hub, https://www.science.org/content/article/new-crop-covid-19-mrna-vaccines-could-be-easier-store-cheaper-use, https://ClinicalTrials.gov/show/NCT05231369, https://ClinicalTrials.gov/show/NCT05605470, http://creativecommons.org/licenses/by/4.0/.

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