The positive cut-off was the subtracted OD450+3SD. Tian, J. H. et al. a-0ZG{Px(rA![|-Ml0(9ELO_>+Rf_I4!=fuPq^$\1$j/ In the detection step, staining of the living cells with 0.02% neutral red (Sigma Aldrich, USA) in 1X PBS (Invitrogen, Carlsbad, CA, USA) was used instead of viral protein staining employing anti-nucleocapsid (1:5,000) used in Experiment 1. Fact Sheet for Healthcare Providers Hum Vaccin Immunother 12, 23512356 (2016). 199 0 obj <>stream 8 more], Anti-spike antibody response to natural SARS-CoV-2 infection in the {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 team also determined whether the S1 subunit influences mature neurons during cell exposure. Anti-spike protein to determine SARS-CoV-2 antibody levels: Is there a Moreover, the low dose regimen was also shown to induce a marked reduction in viral load in nasal turbinates, brain, and lung tissues compared to sham-treated controls. This was followed by exposure to the same S1 concentration for seven consecutive days. The RNAscope ISH assay was performed using an RNAscope 2.5 HD Red Detection Kit (ACD, 322372) as followed. Understanding Your Spike Protein Results | CityMD Centrifuge GOLD SST tube and route to Eastlake Virology (EVIR rack 81). These results reflect the real S protein dynamic as shedding of S1 could be detected in viral infection33,34. SARS-COV-2 Variants: Differences and Potential of Immune Evasion. Science 377, 890894 (2022). Thank you for visiting nature.com. COVID19 Vaccine Tracker [cited 2022 19 August]. 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. The 4-week gap was used according to the preclinical study protocol of ChAdOX-vectored vaccines65,66. ChulaCov19 vaccination could provide 100% protection from severe clinical signs and mortality in mice. Please use one of the following formats to cite this article in your essay, paper or report: Kunkalikar, Bhavana. a Kinetics of total IgG at 2 weeks after receiving 1 or 2 doses of 0.2, 1, 10, and 30g of ChulaCov19. Real-world effectiveness of COVID-19 vaccines: a literature review and meta-analysis. Samples from 69 patients were included in this study. This candidate vaccine has now completed non-clinical toxicity and biodistribution studies and has entered Phase 1 and 2 human trials. Peletta, A. et al. Oran, D. P. & Topol, E. J. The interpretation of the calculated ratios was performed as manufacturer's recommendation. Mice were bled at 2 weeks after each dose and antibody responses were measured by ELISA and/or neutralization assays. Detailed amino sequence was shown in Supplementary File1. This is a surrogate marker indicative of vaccine effectiveness, or the sterilizing immunity as reported in the previous study27. Some must be performed in a laboratory by trained personnel, some can be performed at the point of care, and others can be . According to French regulations, the study was approved by the French ethics committee (Health Data Hub, approval number: F20211217094518). Median time between last vaccination and sampling was 5.2 months (3.16.4). Note; 4 mice in 10g group were analyzed for psVNT50 against BA.4/5 due to the limited volume of serum samples. 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. Each dot represents an individual animal. Homologouse prime/boost results of each vaccine were included. While neurological signs have been documented in some, not all, coronavirus disease 2019 (COVID-19) patients, the precise mechanism by which viruses affect neuronal cells is still unknown and, thus, a subject of investigation. KR, DW, MGA, CK, EP, and SB are co-inventors of the submitted ChulaCov19 mRNA vaccines Patent. No serologic tests for SARS-CoV-2 are approved by the FDA; some, but not all, commercially available serologic tests for SARS-CoV-2 have received EUAs . 2b). 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). ADS These services aid in identifying a relative . showed time-dependent changes in the comparability of different antibody tests with samples collected at different time points [26]. James Heyes, A. J., Kieu Mong, L. A. M., Alan, D. MARTIN. The median values observed for the antibody binding assays were 143 BAU/ml (IQR 39748) for Abbott, 55 BAU/ml (IQR 19217) for Beckman, 636 BAU/ml (IQR 982369) for Roche, and 161 BAU/ml (IQR 32574) for Siemens, which demonstrated the variations between the assays (overall P < 0.0001). This was consistent with the prior study in K18-hACE2 that intranasal inoculation with the similar range of virus caused death within 1 week22. Pharmaceutics 14, 1427 (2022). These viruses adapted to increase the transmissibility, severity and/or immune evasion8. 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. Article PubMed Central 1, 8 (2018). Article When correlating protective efficacy and NAb titers induced by ChulaCov19, a micro-VNT50 titer of 2,560 before challenge in 1 g immunized mice was found to completely prevent viral burden in the lung as analyzed by ISH and RT-qPCR (Figs. p<0.05 and p<0.01 are indicated by * and **, respectively. You are using a browser version with limited support for CSS. Zhang, L. et al. 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. SARS CoV 2 Spike Antibody, IgG Lancet Infect Dis 21, 13521354 (2021). The OD450 of blanks were subtracted from OD450 of each sample before calculating antibody titer. Whether differences in response impact vaccine efficacy needs further study. Baseline characteristics are shown in Table 1. Moreover, anamnestic NAb response is undetectable in challenge animals. mSphere 7, e0024322 (2022). Comparing the clinical efficacy of COVID-19 vaccines: a systematic review and network meta-analysis. Magnitude of asymptomatic COVID-19 cases throughout the course of infection: A systematic review and meta-analysis. SARS2Mutant: SARS-CoV-2 amino-acid mutation atlas database Another limitation was the lack of an external cohort to validate the suggested thresholds. Lancet. Lancet 396, 19791993 (2021). Different studies have used different methods to measure antibody levels, making it difficult to compare results and establish a universal cutoff value conferring protection in immunocompromised patients. Posted in: Medical Science News | Medical Research News | Disease/Infection News, Tags: ACE2, Angiotensin, Angiotensin-Converting Enzyme 2, Antibodies, Antibody, Blood, Blood Pressure, Brain, Cell, Coronavirus, Coronavirus Disease COVID-19, covid-19, Electrode, Enzyme, Frequency, Membrane, micro, Neurons, Newborn, Phenotype, Protein, Receptor, Research, Respiratory, SARS, SARS-CoV-2, Severe Acute Respiratory, Severe Acute Respiratory Syndrome, Spike Protein, Syndrome, Vaccine, Virus. Jairak, W. et al. A. This observation correlates with that of a recent clinical study report53. 4b). Note: tissues from 3/5 animals in control group were collected at day 5. Your Spike Protein Antibody results will be reported as a reference range: >/= 0.80 U/mL: This is a positive result for anti-SARS CoV-2S. 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. PubMed Central In vaccinated people: Nonreactive (Negative, <50.0 AU/mL) results do not rule out SARS-CoV-2 infection, particularly in those who have recently been in contact with the virus. PLOS ONE promises fair, rigorous peer review, Vaccines (Basel) 9, 874 (2021). A vaccine efficacy of 80% was achieved with 264 binding antibody units (BAU)/ml (95% confidence interval [CI]: 108, 806) for anti-spike, and 506 BAU/ml (95% CI: 135, over limit) for anti- receptor-binding domain (RBD) antibodies. PubMed The results demonstrated that, at least up to 12 months, only minor changes were observed when the particles were stored in 75 oC (Supplementary Table1) and were still within the acceptable criteria. As required by French regulations, patients attending clinical laboratories are informed that their biological results can be used for research purposes and that they are free to refuse to allow this (information annotated in the clinical laboratory report). 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]. We suggest specific adjusted thresholds (BAU/ml) for the four commercial antibody assays that are used to assess pre-exposure prophylaxis in immunocompromised patients. The S1 subunit interacts with the angiotensin-converting enzyme 2 (ACE2) receptor present in the intestinal and lung cells. Challenge study was conducted in ABSL-3 facility at AFRIMS, Bangkok, Thailand. 5b). Quantitative SARS-CoV-2 anti-spike responses to Pfizer - PubMed SARS-CoV-2 RNA-positive cells were examined and counted unblind by certified personnel. At this time-point, the NAb titers against both Omicron subvariants were still in the same level with week 5 titers (Fig. 3a). Negative test results do not rule out the possibility of an infection with SARS-CoV-2. ROC curves for each antibody binding assay according to Genscript sVNT. 4a). For example, for 10g dose group, the GMTs of psVNT50 for Delta (B.1.617.2) and Omicron (BA.1) variants decreased 5.9 and 14.3 folds when compared against WT (Wuhan-Hu1) strain, respectively (Fig. Goat-anti-mouse IgG-FITC, donkey-anti-rabbit IgG-FITC (both were from BioLegend, CA, USA) or goat-anti-human AlexaFluor647 (Southern Biotech, AL, USA), at dilution of 1:5,000 was used as secondary antibodies following anti-RBD, -S1, -S2 or PCS staining. Indeed, cutoff values established using commercially available SARS CoV-2 diagnostic antibodies cannot represent a gold standard threshold value related to a level of neutralizing activity. endstream In the present study, researchers quantified the neurological phenotypes induced in neurons by the SARS-CoV-2 S protein. Lv, J., Wu, H., Xu, J. World Health Organization. For the Siemens assay, the optimal cutoff was within the same range as the reference cutoff (270 BAU/ml). Further information on research design is available in theNature Portfolio Reporting Summary linked to this article. 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). N Engl J Med 383, 26032615 (2020). The reaction was carried out employing T7 RNA polymerase (MegaScript, ThermoFisher Scientific, MA, USA) on a linearized plasmid (Not I/Afl II double digestions). Feikin, D.R. Thus, in this study, vaccine-induced disease enhancement is less likely as demonstrated by the Th1-oriented response (Fig. 2023. The S protein trimer (S-trimer), depicted in Fig. Samples from 69 patients were analyzed. These tests should not be used to diagnosis or exclude acute SARS-CoV-2 infection. Her educational background allowed her to foster an interest in anatomical and physiological sciences. First bivalent COVID-19 booster vaccine approved by UK medicines regulator). Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons. Koonpaew, S. et al. Characteristics like the number of bursts per electrode, their duration, frequency, and the number of spikes per burst according to the treatment condition were also quantified. 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. 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. Laboratoire AlphabioBiogroup, Marseille, France, Affiliation: Interpretation of SARS-CoV-2 Immune Response Tests 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. This program is a strong foundation for the fight against the next pandemic by increasing preparedness to make mRNA vaccine widely and timely accessible for LMICs, including Thailand. Single-dose administration and the influence of the timing of the booster dose on immunogenicity and efficacy of ChAdOx1 nCoV-19 (AZD1222) vaccine: a pooled analysis of four randomised trials. % The mRNA was transcribed to contain 101 nucleotide of adenine (101-poly(A) tails). 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. Native-like SARS-CoV-2 Spike Glycoprotein Expressed by ChAdOx1 nCoV-19/AZD1222 Vaccine. Although the currently available vaccines do not completely prevent infection, they are efficacious in reducing severe symptoms of infected individuals11. Route to Eastlake Virology (EVIR rack 81). CAS PLoS ONE 18(4): As previously observed by Perkmann et al. : data collection, A.T., A.J., K.R., K.P., T.P., M.R., D.W., and K.R. Quantitative SARS-CoV-2 anti-spike responses to Pfizer-BioNTech and Oxford-AstraZeneca vaccines by previous infection status SARS-CoV-2 vaccination leads to detectable anti-spike antibodies in nearly all adult HCWs. 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. Experiment 1: dose-response of homologous ChulaCov19 prime/boost study, mice were immunized twice intramuscularly at 3 weeks interval of ChulaCov19 with dosage ranging from 0.2, 1, 10, to 30g. endobj Immunofluorescent results mostly correlate with PCR data. Protective activity of mRNA vaccines against ancestral and variant SARS-CoV-2 strains. The primary components of the SARS-CoV-2 structure are envelope (E), spike (S), membrane (M), and nucleocapsid (N) proteins. In all vaccinated groups, the number of spots that were detected after peptide pool #3-5 and pool #9 stimulation were 7484% and 810%, respectively (Fig. : draft manuscript preparation. a mice were immunized with various doses of ChulaCov19 analyzed at 2 weeks after the second dose. . Nucleoside-modified mRNA was produced by in vitro transcription (IVT) by substitution of uridine triphosphate (UTP) with N1-methylpseudouridine (m1) triphosphate (TriLink, Biotechnologies, San Diego, CA, USA), detailed elsewhere58. Int J Infect Dis 112, 227234 (2021). Source data are provided as a Source Data file. Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. Spin within 24 hours and prior to shipment. 2563.1/8 and 2564.1/4, National Research Council of Thailand NRCT. JAMA Netw Open 4, e2137257 (2021). The ethics committee waived the need for formal written informed consent from patients, as this study was performed on clinical data retrieved from routine tests; thus, no patient was specifically included in this study. In these preclinical studies in mice, we have demonstrated that ChulaCov19, a secreted, prefusion non-stabilized ectodomain spike mRNA vaccine, elicited robust Spike-specific antibody and T-cell responses which has also translated into efficacy in protecting transgenic mice from SARS-CoV-2. 007/2563), and the Armed Forces Research Institute of Medical Sciences, AFRIMS (IACUC approval no. (2023, April 27). COVID-19 CORONAVIRUS PANDEMIC [updated 19 August 2022; cited 2022 19 August 2022]. Google Scholar. This initiative is ready to be part of the global effort to make mRNA vaccines more quickly and widely available when facing new variants or the next pandemic. Peletta, A. et al. 6b. Nature 608, 593602 (2022). (2023) Anti-spike protein to determine SARS-CoV-2 antibody levels: Is there a specific threshold conferring protection in immunocompromised patients? Within the brain, ACE2 is predominantly expressed in the brain stem and regions whose primary function is to regulate blood pressure and cardiovascular function. Article 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. 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.