Then, HRP-conjugated secondary antibodies, including rabbit anti-mouse IgG, dilution 1:10,000 (KPL, MD, USA), -IgG1 (dilution 1:5000), or -IgG2a dilution 1:5000 (both were from Southern Biotech, AL, USA) were added for an additional 1h. After washing, the signals were detected by adding tetramethylbenzidine (TMB) substrate (BioLegend, San Diego, CA, USA). SARS-CoV-2 is the name of the virus that causes coronavirus disease 2019 (COVID-19). Some tests provide results rapidly (within minutes); others require 1-3 days for processing. Nature 589, 603607 (2021). between patient and physician/doctor and the medical advice they may provide. 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]. 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. Recombinant S protein with abolished S1/S2 cleavage site was used as positive control in HEK293T-hACE-2 binding assay (right panel of 2b) and western blot (right lane of each panel in 2c). All samples were collected at the Alphabio Laboratory in Marseille, France (European Hospital, AlphabioBiogroup). PLoS One 16, e0248007 (2021). Lancet 396, 467478 (2020). IgG2a and IgG1 subclasses were also assessed to determine Th1 and Th2 responses, respectively. 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. Moreover, the feasibility of large-scale production as well as rapid adaptability to new variants are major advantages of the mRNA production platform. Labcorp offers both semi-quantitative and qualitative options for testing for spike protein antibodies. A Single-Cycle Influenza A Virus-Based SARS-CoV-2 Vaccine Elicits Potent Immune Responses in a Mouse Model. The S protein trimer (S-trimer), depicted in Fig. Results from antibody testing should not be used as the sole basis to diagnose or exclude SARS-CoV-2 infection or to inform infection status. Derby, M., Alexander-Miller, M., Tse, R. & Berzofsky, J. High-avidity CTL exploit two complementary mechanisms to provide better protection against viral infection than low-avidity CTL. The study also noted that the RBD may be accountable for the suppression of neuronal signals. Vacharathit, V. et al. p<0.05 and p<0.01 are indicated by * and **, respectively. WIPO (2020). E.P., C.K., and K.R. On the basis of these data at present anti-SARS CoV-2 serological assays' results are not interchangeable, and, more importantly, individual immune monitoring should be performed with the same method. 3b). By Day 4 after challenge, two mice in PBS-receiving group (control) began to show clinical signs of anorexia, lethargy, and rough hair coat. There were few limitations in this study. on this website is designed to support, not to replace the relationship Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. 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. although all assays showed good agreement with the Genscript sVNT, they were not interchangeable, even when converted to BAU/ml [10]. Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike. In the clinical setting, >8 weeks interval for AZD1222 was recommended to maximize the vaccine efficacy52. The bands corresponding to S1, S2 and intact S (S0) were detected. 6c. Tissues were collected at week 5+6 days for assessment of viral RNA. The team also determined whether the S1 subunit influences mature neurons during cell exposure. This is consistent with a previous report46. Laboratoires Oriade NovialeBiogroup, Grenoble, France, Affiliation: Therefore SARS-CoV-2 serology may be standardized. Further investigation using different techniques, such as viral isolation and titration from the collected tissues is required to draw a definite conclusion. World Health Organization. In this study, the S1 and S2 subunits of the spike protein . Protection of K18-hACE2 mice and ferrets against SARS-CoV-2 challenge by a single-dose mucosal immunization with a parainfluenza virus 5-based COVID-19 vaccine. Common SARS-CoV-2 virus antigenic targets include spike, envelope, and nucleocapsid proteins [1]. To test the hypothesis that the S1 receptor-binding domain (RBD) may be the reason for burst reduction, the team collected and assessed purified recombinant RBD. In contrast, ChulaCov19 immunized mice, both 1g and 10g doses enabled 100% survival compared to full mortality rate in PBS-immunized mice. The female/male ratio was 67/33, and the median age was 47 years (IQR 3463). Copyright: 2023 Halfon et al. N Engl J Med 383, 26032615 (2020). ROC curves for each antibody binding assay according to Genscript sVNT. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Kunkalikar, Bhavana. 01 May 2023. After 1h incubation at 37C, plates were washed vigorously with washing buffer (PBS+0.5% Tween 20, PBST). Protection against COVID-19 is thought to depend on the presence of specific antibodies against the virus, as well as the function of other components of the immune system such as T cells. Proc Natl Acad Sci U S A 93, 41024107 (1996). 007/2563), and the Armed Forces Research Institute of Medical Sciences, AFRIMS (IACUC approval no. Safety and Immunogenicity of Two RNA-Based Covid-19 Vaccine Candidates. This is a surrogate marker indicative of vaccine effectiveness, or the sterilizing immunity as reported in the previous study27. Pharmaceutics 14, 1427 (2022). Together with the emergence of new VOCs, a booster dose (either homologous or heterologous vaccine modality) is required to enhance the vaccine effectiveness15. 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. In the immunogenicity dose-response and prime/boost studies (Experiment 1 and 2), NAb measurement was carried out as previously described56,68 based on live-virus micro-VNT50 against WT (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), Delta (B1.617.2) variants in VERO E6 cells with positive cut-off of 1:20. Most convalescent patients tested with Tspot are reactive depending on which antigen is tested and which technique is used. While most serologic assays are qualitative, a quantitative serologic . By using immunofluorescent assay, employing RBD-, S1-, S2-specific antibodies or PCS, the S proteins were observed within the cytoplasm of transfected cells while untransfected cells were negative for fluorescent signal (Fig. Frdrique Retornaz, Hence, the low micro-VNT50 titer in the homologous AZD1222 group might increase if the interval between each dose is longer than 4 weeks as used in this study. Boxplots for each antibody binding assay according to Genscript sVNT positive and negative results. K18-hACE2 transgenic mice are highly susceptible and displayed clinical signs following SARS-CoV-2 challenge22,23. 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 Surrogate Virus Neutralization Test (sVNT) Kit (cPass) was purchased from Genscript (Piscataway, USA). Many types of tests are used to detect SARS-CoV-2, 1 and their performance characteristics vary. All data were fully anonymized before the analysis. 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. ADS SARS-CoV-2 delta variant infection in domestic dogs and cats, Thailand. broad scope, and wide readership a perfect fit for your research every time. 6c) may be due to RT-qPCR, a highly sensitive method detecting the free viral RNA from disintegrated virus. Covid-19 Tracker: More Than 12.6 Billion Shots Given [updated 31 August 2022; cited 2022 31 August]. as a booster dose in mice that had been primed with CoronaVac or AZD1222 (Experiment 2). 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. At week 5 (2 weeks after the second dose), all mice in both vaccinated groups showed increased NAb levels. The analysis of the responses to different parts of S-specific pool peptides in all vaccinated groups showed that peptide pool #3-5 (which include receptor-binding domain or RBD) and pool #9 (which includes Heptad Repeat 2 or HR2) in S1 and S2, respectively, were the most common peptides pools recognized by the vaccinated mice T-cells. After 2 doses of ChulaCov19 or phosphate-buffered saline (PBS, control group) with a 3-week interval, K18-hACE2 mice were tested for NAb kinetics against live SARS-CoV-2 strain hCoV-19/Hongkong/VM20001061/2020. Stability: Sample stable off the clot, red blood cells, or separator gel for 7 days at 2-8C. Chen, X. et al. Figures were created with BioRender.com. DNA Vaccine Administered by Cationic Lipoplexes or by In Vivo Electroporation Induces Comparable Antibody Responses against SARS-CoV-2 in Mice. On day zero, neurons obtained from newborn P1 mice were treated with recombinant SARS-CoV-2 S protein and S1 and S2 S2 subunits. Peletta, A. et al. ];V^srE]DwCyTPre_fyG;Cb@*\d$ j '-|,!]jF/J9r\s%3! The mRNA vaccine technology transfer hub [updated 21 June 2022. In addition, the pseudovirus neutralization test (psVNT50) against lentiviral pseudovirus bearing a codon-optimized spike gene, described previously69,70, was also used for determination of the neutralizing activity against WT, (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), Delta (B1.617.2), and Omicron (B1.1.529; BA.1 and BA.4/5 subvariants) variants. In addition, there was no anamnestic antibody response detected in the ChulaCov19 vaccinated mice after viral challenge (Fig. For western blot analysis, cell culture supernatant was analyzed by 12% polyacrylamide gel then transferred onto nitrocellulose membrane. Secreted S protein was also subjected for analysis of its binding capability to hACE2. Google Scholar. The geometric mean titers (GMTs) of NAb against wild-type (WT, Wuhan-Hu1) virus are 1,280, 11,762, 54,047, and 62,084, respectively. CAS The positive cut-off was the subtracted OD450+3SD. It also markedly reduced viral RNA burden in serum and tissues. In Experiment 3, the durability of NAb induced by ChulaCov19 was monitored until week 18 (15 weeks after the 2nd dose). The National Library of Medicine is running a pilotto include preprints that result from research funded by NIH in PMC and PubMed. Helmy, Y. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. 1, 8 (2018). 7, eabi5246 (2021). tG gh3o^r3^Z\PQE5)?xLr3El79/iDm}y)MjbV|bBuD7l[U}\Cxo Science 368, 489493 (2020). Data are presented as GMT of micro-VNT50 titer with 95% confident interval. Moreover, ChAdOx1: AZD1222 that used unmodified S has been shown to induce high level of NAb and T cells responses even after a single immunization dose in two mouse strains38. Source data are provided as a Source Data file. 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. 4d). Four antibody binding assays were used for serological testing according to the instructions of the manufacturer. Molecular-based testing is used to diagnose COVID-19, and serologic testing of antibodies specific to SARS-CoV-2 is used to detect past infection. Klemis, V. et al. Mol Ther Nucleic Acids 15, 2635 (2019). This study aimed to describe serum-IgG responses to SARS-CoV-2 in a cohort of patients with both severe and mild COVID-19, including extended studies of patients who remained seronegative more than 90 . Within the brain, ACE2 is predominantly expressed in the brain stem and regions whose primary function is to regulate blood pressure and cardiovascular function. Thank you for visiting nature.com. The objective of the present study was to establish a new optimal threshold of protection for four different SARS-CoV-2 antibody assays [14]. Nature Communications (Nat Commun) A Facile Method for the Removal of dsRNA Contaminant from In Vitro-Transcribed mRNA. Biomedicines 10, 1464 (2022). Here, we describe the preclinical studies of ChulaCov19, a SARS-CoV-2 mRNA encoding prefusion-unstabilized ectodomain spike protein encapsulated in lipid nanoparticles (LNP). Immunogenicity and protective efficacy of SARS-CoV-2 mRNA vaccine encoding secreted non-stabilized spike in female mice. Challenge study was conducted in ABSL-3 facility at AFRIMS, Bangkok, Thailand. \1;nJ/mjJ=DqXlU,u>z}x)tU#K>/#}idN"%W$YoSA14Ys5+VlE5-3a+`h"xD%5n#L$\g%[&0Gy-x;a>$'+6#am#WK>nxW|^E~YS t4G2G9V$Mf=E5y? Global Percentage of Asymptomatic SARS-CoV-2 Infections Among the Tested Population and Individuals With Confirmed COVID-19 Diagnosis: A Systematic Review and Meta-analysis. EP was also supported by Faculty of Medicine, Chulalongkorn University, grant No. World Health Organization. 4e). SARS-CoV-2 Antibodies (NCVIGG, NCVIGQ)[NCVIGB], The qualitative detection of anti-Nucleocapsid IgG (NCVIGG) and the quantitative detection of anti-Spike IgG (NCVIGQ) antibodies. 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. In brief, mouse splenocytes at 5105 cells/well were cultured with SARS-CoV-2 spike peptide pools spanning the entire sequence of spike protein, 25 peptides/pool (Mimotopes, Mulgrave, Victoria, Australia) at a final concentration of 2g/mL at 37C, 5% CO2 for 40h. Pools 15 and 610 corresponded to S1 and S2 regions of spike protein, respectively. Funding: The author(s) received no specific funding for this work. As the Omicron subvariant BA.4/5 is currently spreading worldwide, we have also assessed cross-neutralization and found that the NAb GMT measured by psVNT50 against BA.4/5 in homologous ChulaCov19 vaccination or heterologous boosted with ChulaCov19 groups were significantly better than either of the CoronaVac or AZD1222 homologous vaccination (Fig. More info. The overall concordance between the antibody binding assays and the Genscript sVNT also increased consistently i.e., 11% increase for Roche (86% concordance), 10% increase for Beckman (88% concordance), 2% increase for Siemens (90% concordance), and 1% increase for the Abbott assay (88% concordance). Pathogenesis of SARS-CoV-2 in Transgenic Mice Expressing Human Angiotensin-Converting Enzyme 2. COVID-19 CORONAVIRUS PANDEMIC [updated 19 August 2022; cited 2022 19 August 2022]. Her college project work based on The manifestations and causes of sickle cell anemia formed the stepping stone to a life-long fascination with human pathophysiology. How are Women Emotionally Affected After a Hysterectomy? Prevalence of Asymptomatic SARS-CoV-2 Infection. Article PubMed J. Clin. doi:10.1371/journal.pone.0281257, Editor: Deniz Can Guven, Elazg Fethi Sekin City Hospital: Elazig Fethi Sekin Sehir Hastanesi, TURKEY, Received: November 17, 2022; Accepted: January 18, 2023; Published: April 28, 2023. Heat-inactivated mice sera that were diluted 1:100 was added in duplicates into RBD-coated wells and incubated at RT for 2h. Then, either goat-antimouse IgG-HRP (1:40,000 dilution, KPL, USA) or goat-anti-mouse IgA-HRP (1:10,000 dilution, KPL, USA) was added to each well (100l/well) and incubated at RT for 1h. The peroxidase reaction was visualized by adding Sureblue TMB solution (KPL, USA) and incubating in the dark at RT for 15 and 20min for IgG and IgA ELISAs, respectively. Department of Infectious Diseases and Internal Medicine, Hpital Europen, Marseille, France, Affiliation: These factors might make it difficult to draw a strong conclusion on vaccine efficacy from the current of experiments. Alexander-Miller, M. A., Leggatt, G. R., Sarin, A. Developing highly effective vaccine platforms like mRNA technology in low- and middle-income countries (LMICs) is therefore an important goal21. Int J Infect Dis 114, 252260 (2022). 199 0 obj <>stream Bowen, J. E. et al. PubMed Central Baseline characteristics are shown in Table 1. 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. Before administering S1 to neurons on day zero, a human monoclonal anti-S1 antibody was sampled and neutralized using the antibody. Nl5AMM(&R/ You are using a browser version with limited support for CSS. This contrasts with SARS CoV-1 where peak viral shedding occurs after patients were already quite ill5,6. Guillaume Penaranda [ view less ], Affiliations: Her academic background is in Pharmaceutical sciences and she holds a Bachelor's degree in Pharmacy. Statistical significance was determined by two-sided MannWhitney test. 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. The most effective COVID-19 vaccines are mRNA-based and were first approved in the United Kingdom, the United States, and Europe. Comparing the clinical efficacy of COVID-19 vaccines: a systematic review and network meta-analysis. PubMed Five micrograms of ChulaCov19 was selected as we aimed to standardize the dosage to 1/10 of human dose for all vaccines (50g per dose of ChulaCov19 was used in phase II studies, Clinical Trial Identifiers: NCT05231369 and NCT05605470)63,64. From nanoparticle-based enrichment to mass spec refinements, they explore how these tools facilitate unbiased, deep, and rapid proteomics. The test can provide information about how your body reacted to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). All authors reviewed the results and approved the final version of the manuscript. d psVNT50 NAb titer results at two weeks after the second dose in various prime/boost regimens againt Omicron BA.1 and BA.4/5 subvariants. 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. However, the slightly higher level compared to the 30-g group was not statistically significant. Experiment 1: (a) Live-virus microneutralization (micro-VNT50) titers against WT (Wuhan-Hu1) live-virus at two weeks after receiving each vaccine dose. Available from: https://covid19.trackvaccines.org/agency/who (2022). Med. The average decline from peak to euthanasia among PBS-receiving mice was 17%. After SARS-CoV-2 challenge, there was no measurable decline in body weight among vaccinated groups. However, at week 2 after the first dose, 6/6 and 4/6 animals from the 10g and 1g groups, respectively, showed a dose-dependent manner of NAb response to vaccine administration. The nonparametric KruskalWallis test for multiple comparisons was used to compare all assays. 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. The S1 subunit substantially lowered the number of bursts per electrode, whereas the S2 subunit did not exhibit the same degree of reduction. In contrast, sham-treated animals failed to show any NAb response except for one animal on Wk5+6d (Fig. https://solidarites-sante.gouv.fr/IMG/pdf/cosv_-_recommandations_pour_la_protection_des_personnes_severement_immunodeprimees_-_19_novembre_2021.pdf, https://www.who.int/publications/m/item/WHO-BS-2020.2403, Corrections, Expressions of Concern, and Retractions. The VITROS Immunodiagnostic Products Anti -SARS-CoV-2 Total test is performed using the VITROS Anti -SARS- CoV-2 Total Reagent Pack and the VITROS Anti -SARS-CoV-2 Total. 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). Seventeen female K18-hACE2 mice (B6.Cg-Tg(K18-hACE2)2Prlmn/J), 7 weeks old (The Jackson Laboratory, Bar Harbor, ME, USA) were randomly divided into 3 groups. Gilles Antoniotti, Baseline NAb levels at week 0 of all mice were negative. Philippe Cartlamy, 4c). Use the Previous and Next buttons to navigate the slides or the slide controller buttons at the end to navigate through each slide. Vacharathit, V. et al. These common antibody tests use purified proteins of SARS-CoV-2 (not a live virus) to detect the presence of binding antibodies that attach to a virus, per the CDC. m8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p a8 0 H $g 3 @p 18n e>0> Sci Transl Med 14, eabm3302 (2022). Cohen J. Publishers note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Indeed, antibody therapy for pre-exposure prophylaxis (PrEP), may be efficient in preventing hospitalization in immunocompromised patients, regardless of the variant involved. J Control Release 217, 345351 (2015). The goal of experiment 2 was to assess the potential role of ChulaCov19 as a booster in a setting of heterologous primed with other COVID-19 vaccine platforms. Regarding the vaccine construct characterization, protein expression studies revealed S proteins were expressed both in intracellular and extracellular compartments when detected either by specific antibodies or patient sera (Fig. It was also evaluated for the protective efficacy in transgenic mice expressing human angiotensin-converting enzyme-2 (ACE2), Fig. https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. : study conception and design, E.P., K.T., and C.K.
Nightstand With Charging Port, Rapid Fire Pellet Rifle, Dunwoody Police Detective, Ronnie Barnes Obituary, Kelly Educational Staffing Hillsborough County, Articles A