Magnitude of asymptomatic COVID-19 cases throughout the course of infection: A systematic review and meta-analysis. The second dose of ChulaCov19 strongly augmented the IgG antibody levels with an increase of 10-19 folds, p<0.01 for all dose ranges (Fig. For tissue samples, RNeasy Mini Kit (QIAGEN, Hilden, Germany) was used following manufacturer instructions. The RT-qPCR data showed that both doses of vaccine prevented the expression of SARS-CoV-2 viremia at 5 or 6 days after viral inoculation. Baseline characteristics are shown in Table 1. A Thermostable mRNA Vaccine against COVID-19. Get the most important science stories of the day, free in your inbox. Kim, H. W. et al. Vacharathit, V. et al. Source data are provided as a Source Data file. 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. 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. https://doi.org/10.1038/s41467-023-37795-0, DOI: https://doi.org/10.1038/s41467-023-37795-0. Vaccines (Basel) 9, (2021). World Health Organization. Vaccines (Basel) 9, 874 (2021). The reaction was stopped by adding 50l/well of 0.5M sulfuric acid. PubMed Central 2a). ADS Establishment of an mRNA vaccine platform in low- and middle-income countries (LMICs) is important to enhance vaccine accessibility and ensure future pandemic preparedness. Detailed amino sequence was shown in Supplementary File1. Moreover, anamnestic NAb response is undetectable in challenge animals. Pharmaceutics 14, 1427 (2022). between patient and physician/doctor and the medical advice they may provide. These results reflect the real S protein dynamic as shedding of S1 could be detected in viral infection33,34. Objectives: The aim was to determine the antibody response against SARS-CoV-2 spike protein and nucleoprotein using four automated immunoassays and three ELISAs for the detection of total Ig antibodies (Roche) or IgG (Abbott, Diasorin, Snibe, Euroimmun, Mikrogen) in COVID-19 patients. The purified mRNA-S (ChulaCov19) with undetectable endotoxin was tested for protein expression in VERO E6 cells. Prevention CfDCa. SARS-CoV-2 neutralizing antibodies decline over one year and patients with severe COVID-19 pneumonia display a unique cytokine profile. Post-translational modifications were also similar to those observed on SARS-CoV-241. Median time between last vaccination and sampling was 5.2 months (3.16.4). SARS-CoV-2 is the name of the virus that causes coronavirus disease 2019 (COVID-19). By submitting a comment you agree to abide by our Terms and Community Guidelines. Serum-IgG responses to SARS-CoV-2 after mild and severe COVID-19 - PLOS Splenocytes from mice immunized with various dosages of ChulaCov19 (Experiment 1) were analyzed as summed frequency of S-specific IFN- positive T cells (Fig. However, this was still far lower than using homologous ChulaCov19 or AZD1222-prime/ChulaCov19-boost immunization regimens (Fig. p<0.05 and p<0.01 are indicated by * and **, respectively. Front Immunol 11, 1673 (2020). In the heterologous vs homologous prime/boost experiment (Experiment 2), homologous ChulaCov19 and homologous AZD1222 immunizations elicited comparable levels of S-specific IFN- positive T cells responses which was 2482 and 2210 SFC/106 splenocytes, respectively. At week 18, the NAb against WT (Wuhan-Hu1) and Delta (B.1.617.2) decreased approximately 2-fold but not statistically significant when compare with week 5 titers. Limited and Short-Lasting Virus Neutralizing Titers Induced by Inactivated SARS-CoV-2 Vaccine. DNA Vaccine Administered by Cationic Lipoplexes or by In Vivo Electroporation Induces Comparable Antibody Responses against SARS-CoV-2 in Mice. Vaccines (Basel) 10, 613 (2022). This was followed by exposure to the same S1 concentration for seven consecutive days. {KnXEW;>2THg_J}iX,n7 UndO'%vh9(WG(Rf&oKnn>*&j6$79^*G$73sxv_7$wWfbgD7l7`{ FD5`yK]TS.t0 bM/.<1~ Na RUL6>lnn;P"_1m^ The GMT of micro-VNT50 titers at week 5 were 15,343 and 4424 in the 10 g and 1 g groups, respectively, p=0.0325. 9, 1225 (2020). The results should always be assessed in conjunction with patient's medical history, clinical presentation, and other findings. The S-specific total IgG after 1 or 2 doses of ChulaCov19 was analyzed in mice sera from experiment 1. 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 In the control group, 3 out of 5 mice reached euthanasia criteria on Day 5 hence only 2 mice were analyzed for body weight on Day 6 after challenge. LMICs received these vaccines much later and in shorter supply, as evidenced by the most recent statistic (as of 31 August 2022) that in several African countries less than 30% of the population has received at least one vaccine dose20. Nature 586, 578582 (2020). Philippe Halfon, ISSN 2041-1723 (online). Having more antibodies means your body can fight infection better than having fewer antibodies. SD; standard deviation. KL and JH are employees of Genevant Sciences Corporation and are named on patent describing lipid nanoparticles. In the present study, researchers quantified the neurological phenotypes induced in neurons by the SARS-CoV-2 S protein. Omicron spike function and neutralizing activity elicited by a comprehensive panel of vaccines. Previous B cell depletion correlated with anti-SARS-CoV-2 IgG levels. However, the slightly higher level compared to the 30-g group was not statistically significant. Mol Ther 28, 15691584 (2020). SARS-CoV-2, SARS-CoV, and MERS-CoV viral load dynamics, duration of viral shedding, and infectiousness: a systematic review and meta-analysis. Helmy, Y. Comparisons were performed between Genscript sVNT positive and negative samples according to antibody binding assays, all of which were significant (P < 0.0001) (Fig 1). Zheng, J. et al. Negative test results do not rule out the possibility of an infection with SARS-CoV-2. At this time-point, 10g dosed mice induced significantly higher in GMTs of micro-VNT50 titers than 1g dosed mice (p=0.0065). Prediction of long-term kinetics of vaccine-elicited neutralizing antibody and time-varying vaccine-specific efficacy against the SARS-CoV-2 Delta variant by clinical endpoint. Calculations were performed using the SAS V9.4 software (SAS Institute Inc., Cary, NC, USA). Her educational background allowed her to foster an interest in anatomical and physiological sciences. x2- p` ` \0`0e`X{StAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA^@%&D7(mylkv IVT mRNA was analyzed on agarose for determination of its integrity. Centrifuge GOLD SST tube and route to Eastlake Virology (EVIR rack 81). How are the results reported for the anti-nucleocapsid antibody test, and what is the clinical significance? Nature 589, 603607 (2021). mSphere 7, e0024322 (2022). Zheng, C. et al. WHO. More info. S1 neutralized by antibodies did not result in a significant decrease in burst activity compared to the control, whereas the conventional S1 treatment on day zero did reduce burst activity. The procedure of mouse IFN- ELISPOT used in this study was described in our previous reports56,72. Quantitative Measurement of Anti-SARS-CoV-2 Antibodies: Analytical and Using western blot, the S protein could be detected in cell culture supernatant when using anti-RBD, -S1, -S2 and PCS as primary antibodies. In a recent study posted to the bioRxiv* preprint server, researchers explored the association between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and burst activities in neurons. The study suggested that S1 is responsible for decreasing burst activities of neuronal populations when cells are exposed early in the course of development. 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. Prefusion RSV F Immunization Elicits Th2-Mediated Lung Pathology in Mice When Formulated With a Th2 (but Not a Th1/Th2-Balanced) Adjuvant Despite Complete Viral Protection. 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. a mice were immunized with various doses of ChulaCov19 analyzed at 2 weeks after the second dose. This study also indicated that neutralizing S1 restores neuronal discharge activities to control levels. The titers were determined in duplicate assays from control (n=5) or vaccinated groups (n=6), respectively. b hACE-2 binding assay (merged): culture supernatant collected from ChulaCov19 transfected cells incubated with HEK293T- hACE-2 cells. In summary, this mRNA vaccine development is an effort to set up the technology platform in LMICS. PLoS One 16, e0248007 (2021). Background Identifying a specific threshold level of SARS-CoV-2 antibodies that confers protection in immunocompromised patients has been very challenging. Each dot represents an individual animal. 1b). Indeed, the BAU/ml values were performed only on the B.1.1.7 variant in neutralization assays and not on different strains of the virus; hence, there may be no relation between immune markers and disease outcome [7]. Baiersdorfer, M. et al. Testing for SARS-CoV-2 Infection. Kunkalikar, Bhavana. Experiment 2: a prime/boost regimen of 5g of ChulaCov19 and 1/10 of human dosage of approved vaccines available during the study period, including viral-vectored (ChAdOx1; AZD1222, Lot A10062, Nonthaburi, Thailand) and inactivated (CoronaVac, Lot C202105081, Beijing, China) vaccines. SARS-CoV-2 Spike Protein Reduces Burst Activities in Neurons Measured by Micro-Electrode Arrays, Omicron spike N679K mutation acts as a loss-of-function mutation attenuating SARS-CoV-2 in vitro & in vivo, The virological characteristics of XBB.1.16. 4c. Therefore, we suggest specific BAU/ml adjusted thresholds for the four commercial antibody assays (Abbott, Beckman, Roche, and Siemens), which can be used to guide the use of PreP in immunocompromised patients. Overall, all assays showed good agreement with the Genscript sVNT. 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. wxWd~{Trru%m#97Z=}<8boK.3E@KT>1oW#!7q%7uJ?IC5 .iM!. Tian, J. H. et al. The mRNA vaccine technology transfer hub [updated 21 June 2022. Cite this article. By Day 4 after challenge, two mice in PBS-receiving group (control) began to show clinical signs of anorexia, lethargy, and rough hair coat. After SARS-CoV-2 challenge, there was no measurable decline in body weight among vaccinated groups. It may also mean your body's immune system has generated a response to a prior COVID-19 infection. This observation correlates with that of a recent clinical study report53. 5a). Statistical analysis was performed using GraphPad Prism 9.0 software (San Diego, CA, USA). Lancet 397, 881891 (2021). 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). Experiment 1: (a) Live-virus microneutralization (micro-VNT50) titers against WT (Wuhan-Hu1) live-virus at two weeks after receiving each vaccine dose. Viruses were propagated in Vero E6 cells to generate sufficient titers 100TCID50 for the micro-VNT50 assay. One-day-old Vero E6 cells were used for measuring the level of neutralizing antibodies by live-virus micro-neutralization (micro-VNT50). 5b). Two approved mRNA vaccines, ComirnatyTM by Pfizer/BioNTech and SpikevaxTM by Moderna, comprise 2 proline substitutions at residues 986 and 987 of the S-protein (known as S-2P) to stabilize the prefusion conformational structure. Currently there are at least 11 approved vaccines using various technology platforms, including mRNA, inactivated virus, viral-vector and recombinant protein10. Note; the IgG2a/IgG1 ratio of 10g and 30g immunized mice were not analyzed due to limited volume of serum samples. Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine. Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. Sci Rep 12, 8403 (2022). Safety and Immunogenicity of ChulaCov19 BNA159 mRNA Vaccine.). 7, eabi5246 (2021). At week 22, the psVNT-50 GMT for WT (Wuhan-Hu1), Delta (B.1.617.2), BA.1 and BA.4/5 were 25,539, 10,722, 2133, and 1707, respectively; 13-57 folds increase from the pre-boost baseline (Week18). INSTRUCTIONS FOR USE CoV2T JAMA Netw Open 4, e2137257 (2021). The Euroimmun Anti-SARS-CoV-2 IgG and IgA tests are separate ELISAs that detect antibodies against the S1 subunit of the SARS-CoV-2 spike protein. Available from: https://www.science.org/content/article/new-crop-covid-19-mrna-vaccines-could-be-easier-store-cheaper-use (2022). Immunization with SARS coronavirus vaccines leads to pulmonary immunopathology on challenge with the SARS virus. The results were compared to the percent inhibition calculated using a functional surrogate of a standardized virus neutralization test (Genscript). Google Scholar. The Abbott Architect SARS-CoV-2 IgG II assay, run under an emergency use authorization from the FDA, is quantitative test designed to detect IgG antibodies to the spike protein of SARS-CoV-2 in serum and plasma from individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection. In many countries, immunization regimens have frequently employed mixtures of different vaccine platforms (also known as a heterologous prime-boost). The data as well as the p values suggested that the anti-S1 antibody reversed the impact of S1 on bursting activities. J Control Release 217, 345351 (2015). Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen. Previous study by Eichinger KM, et al. The possible explanation of the higher detectable viral RNA found in 10 g compared to 1 g immunized mice (Fig. Buschmann, M. D. et al. Bloomberg. These viruses adapted to increase the transmissibility, severity and/or immune evasion8. Article ChulaCov19 was further compared to two approved vaccines (CoronaVac and AZD1222), either in a homologous prime/boost setting or heterologous one (i.e. b Pseudovirus neutralization test (psVNT50) titers at two weeks after the second dose againt WT (Wuhan-Hu1), Delta (B.1.617.2), Omciron (BA.1, and BA.4/5) variants. HEK293T-hACE-2 cells, prepared by transduction of HEK293T cell line with lentiviral habouring hACE-2 gene, used for hACE-2 binding assay was gratefully provided by Dr.Navapon Techakriengkrai55. These authors jointly supervised this work: Drew Weissman, Kiat Ruxrungtham. : data collection, A.T., A.J., K.R., K.P., T.P., M.R., D.W., and K.R. They were widely available in these countries for approximately a year before being accessible on other continents. Pallesen, J. et al. Agreements between antibody-binding assays and Genscript sVNT were performed using Cohens kappa, crude concordance rate, and area under curve (AUC). Laboratoires Oriade NovialeBiogroup, Grenoble, France, Affiliation: Using a serologic test in combination with a NAAT to detect IgG or total antibodies 3 to 4 weeks after symptom onset maximizes the sensitivity and specificity to detect past SARS-CoV-2 infection.