The objective of the present study was to establish a new optimal threshold of protection for four different SARS-CoV-2 antibody assays [14]. Immunogenicity and protective efficacy of SARS-CoV-2 mRNA vaccine encoding secreted non-stabilized spike in female mice, https://doi.org/10.1038/s41467-023-37795-0. Antibody Testing After the COVID-19 Vaccine: What to - CreakyJoints S protein on HEK293T-hACE-2 was stained with anti-RBD, -S1, -S2 or PCS and detected using the same procedure described above. The information of SARS-CoV-2 isolates including, wild-type (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351) and Delta (B.1.617.2) variants for micro-VNT50 assay performed at the Department of Microbiology, Faculty of Sciences, Mahidol University was described previously56,57. Koonpaew, S. et al. There was no detectable viremia in mice in both high or low-dose vaccine-treated groups while an average of 7.71104 GE/mL (ranged from 1.03103 3.75105 GE/mL) of viral RNA was detected in PBS-received mice, Fig. 5b). Sera were collected at weeks 0, 2, 3, 4+6 days, and 5+6 days for NAb measurements. Source data are provided as a source data file. Prompetchara, E., Ketloy, C., Alameh, MG. et al. Philippe Cartlamy, A Multi-Targeting, Nucleoside-Modified mRNA Influenza Virus Vaccine Provides Broad Protection in Mice. At week 3 after dose 1, NAb were still detected in all animals in the 10g group, and 5/6 animals in the 1g group. Detailed amino sequence was shown in Supplementary File1. Therefore, during the surge of Omicron globally, there is a need of a boosting dose even with a first-generation vaccine or ideally with a second-generation vaccine such as a bivalent immunogen containing or encoding of Omicrons spike protein49,50. In vaccinated people: Thank you for visiting nature.com. Differences were considered significant at p<0.05 with exact p-values shown. KR, DW, MGA, CK, EP, and SB are co-inventors of the submitted ChulaCov19 mRNA vaccines Patent. Google Scholar. This contrasts with SARS CoV-1 where peak viral shedding occurs after patients were already quite ill5,6. By continuing to browse this site you agree to our use of cookies. PubMed Central These tests should not be used to diagnosis or exclude acute SARS-CoV-2 infection. Splenocytes from mice immunized with various dosages of ChulaCov19 (Experiment 1) were analyzed as summed frequency of S-specific IFN- positive T cells (Fig. There were few limitations in this study. Google Scholar. 2023. Is there an association between the consumption of ultra-processed food and adverse microbiota-gut-brain axis implications? 01 May 2023. The aim was to assess the threshold of 264 binding antibody units (BAU)/ml using four different SARS-CoV-2 antibody assays (Abbott, Beckman, Roche, and Siemens) and to establish a new optimal threshold of protection for each of the four antibody assays. 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 IgG2a and IgG1 subclasses were also assessed to determine Th1 and Th2 responses, respectively. These results reflect the real S protein dynamic as shedding of S1 could be detected in viral infection33,34. 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). J Immunol 166, 16901697 (2001). The average reduction of viral load in tissues of both vaccine-treated groups relative to the control was 99.9-100%. The team assessed the data using an algorithm devised in-house. The titers were determined in duplicate assays from control (n=5) or vaccinated groups (n=6), respectively. In the control group, positive viral RNA staining was present in individual neurons of the olfactory bulb (4/4), epithelial cells of the nasal sinus (4/5), alveolar epithelial cells and macrophages in the lung (5/5), see Table1. showed time-dependent changes in the comparability of different antibody tests with samples collected at different time points [26]. 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. Jairak, W. et al. 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. mRNA capping was performed by the trinucleotide cap1 analog, CleanCap (TriLink Biotechnologies, San Diego, CA, USA). Guillaume Penaranda Anti-spike antibody response to natural SARS-CoV-2 infection in the 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. 6a). More info. Owned and operated by AZoNetwork, 2000-2023. Centrifuge RED TOP or EDTA tube and aliquot serum/plasma into plastic aliquot tube. In negative control (group 3), 5 mice were immunized with PBS instead of ChulaCov19 using the same schedule. https://doi.org/10.1038/s41467-023-37795-0, DOI: https://doi.org/10.1038/s41467-023-37795-0. Splenocytes were collected at 2 weeks after the second dose (Experiment 1 & 2). Statistical significance was determined by two-sided MannWhitney tests. 2c). This is similar to the previous study of mRNA-1273, which demonstrated that a minimum NAb titer (analyzed by focus reduction neutralization test) of approximately 2,000 was required to completely protect K18-ACE2 mice from ancestral virus with D614G infection32. 5b). Li, R. et al. At this time-point, the NAb titers against both Omicron subvariants were still in the same level with week 5 titers (Fig. 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. BMC Med 20, 36 (2022). All samples were collected at the Alphabio Laboratory in Marseille, France (European Hospital, AlphabioBiogroup). Figures were created with BioRender.com. Optimal cutoffs for distinguishing positivity were calculated using logistic regression on Genscript sVNT binary results (negative/positive), prior to the Youden index maximization approach on receiver operating characteristic curve results. e0281257. Laboratoire AlphabioBiogroup, Marseille, France, The procedure of mouse IFN- ELISPOT used in this study was described in our previous reports56,72. 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. In female BALB/c mice, ChulaCov19 at 0.2, 1, 10, and 30 g elicits robust neutralizing antibody (NAb) and T cell responses in a dose-dependent relationship. Funding: The author(s) received no specific funding for this work. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Differences were considered significant at p<0.05 with exact p-values shown. 1a). Further investigation using different techniques, such as viral isolation and titration from the collected tissues is required to draw a definite conclusion. In contrast, ChulaCov19 immunized mice, both 1g and 10g doses enabled 100% survival compared to full mortality rate in PBS-immunized mice. 4b). T-cell responses to SARS-CoV-2 can be indirectly tested with antigen tests (such as Elispot) that tests for cytokines produced (i.e. https://apps.who.int/iris/handle/10665/363344 (2022).