anti sars cov 2 spike protein test results interpretation anti sars cov 2 spike protein test results interpretation

Mol Ther Nucleic Acids 15, 2635 (2019). 6a). We suggest specific adjusted thresholds (BAU/ml) for the four commercial antibody assays that are used to assess pre-exposure prophylaxis in immunocompromised patients. Laboratoire AlphabioBiogroup, Marseille, France, Affiliation: Supernatant collected from transfected cell was incubated with HEK293T-hACE-2 at 37 oC for 1h then washed twice with PBS. 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. Nat Commun 13, 4710 (2022). JAMA Netw Open 4, e2137257 (2021). In many countries, immunization regimens have frequently employed mixtures of different vaccine platforms (also known as a heterologous prime-boost). Ramasamy, M. N. et al. (accessed May 01, 2023). Quantitative data were reported using median and interquartile range (IQR), and qualitative data were reported using frequency and percentage. Experiments were repeated two times independently with similar results. 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. Among the recently approved vaccines, mRNA modality seems to be the most efficacious as it induces high levels of desired immune responses and protects from severe symptoms16,17. 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. Nat Commun 12, 372 (2021). Vaccines (Basel) 9, 850 (2021). Alene, M. et al. The team then performed a rescue experiment to ascertain if this neuronal phenotype is reversible. Agreements between antibody-binding assays and Genscript sVNT were performed using Cohens kappa, crude concordance rate, and area under curve (AUC). Additional group (Experiment 3) immunized with 5g of ChulaCov19 was included for evaluation of NAb durability as measured at week 18 (15 weeks after received the 2nd dose) and the boosting effect of 3rd ChulaCov19 dose administered at week 20. The neutralizing capacity was estimated by performing a surrogate virus neutralization test (sVNT) assay (GenScript, Piscataway, NJ, USA) as previously described [10,15,16]. Omicron stood out from other variants because it contained mutations that helped it evade immune cell protection. S0 was used to depict unprocessed S protein. J Control Release 217, 345351 (2015). 5a). [ view less ], Affiliations: PubMed Central 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). Do ketogenic diets elevate low-density lipoprotein cholesterol levels? The number of copies of viral RNA per sample was derived from standard curves of serial dilutions of IVTs (5, 50, 5 102, 5 103, 5 104, 5 105 RNA copies number or genomic equivalent (GE)/reaction were included. Therefore, the data indicated that the S1 subunit affected neurons only when the cells were exposed during the earliest stages of development. For the heterologous prime/boost, mice primed with CoronaVac or AZD1222 and then boosted with ChulaCov19 generated significantly higher GMT against WT (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), Delta (B.1.617.2), and Omicron (B.1.529) when compared to the respective homologous prime/boost groups. Copyright: 2023 Halfon et al. Agreement between antibody binding assays and Genscript sVNT positive and negative results according to optimal cutoff values determined for each assay. This research focuses on the impacts of the S protein. 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. The study identified the number of pulses per electrode as the most prominent characteristic that differentiated the spike protein-treated wells from the control wells. Note; the IgG2a/IgG1 ratio of 10g and 30g immunized mice were not analyzed due to limited volume of serum samples. Bars (a) or horizontal lines (b) represent the geometric mean (GMT) for each group while error bars indicate the 95% confident interval. 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. Image Credit: whitehoune/Shutterstock.com. Study: SARS-CoV-2 Spike Protein Reduces Burst Activities in Neurons Measured by Micro-Electrode Arrays. The purified mRNA-S (ChulaCov19) with undetectable endotoxin was tested for protein expression in VERO E6 cells. If testing will be delayed more than 7 days store at -20C or colder. RBD-VLP Vaccines Adjuvanted with Alum or SWE Protect K18-hACE2 Mice against SARS-CoV-2 VOC Challenge. The authors acknowledge all the members of the Chula VRC for their input and support. 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). The results of available anti-SARS-CoV-2 IgG antibody tests may be interpreted in the following way: In a person never vaccinated: Testing positive for antibody against N (nucleocapsid protein), S (spike protein), or RBD (receptor-binding domain of S protein) indicates prior infection. Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons. Feikin, D.R. Background To accurately interpret COVID-19 seroprevalence surveys, knowledge of serum-IgG responses to SARS-CoV-2 with a better understanding of patients who do not seroconvert, is imperative. 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. 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. 2a). They were widely available in these countries for approximately a year before being accessible on other continents. The absorbance was measured at a wavelength of 450nm using a Varioskan microplate reader (ThermoFisher Scientific, Vantaa, Finland). 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. Prompetchara, E. et al. By Day 4 after challenge, two mice in PBS-receiving group (control) began to show clinical signs of anorexia, lethargy, and rough hair coat. 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. A Multi-Targeting, Nucleoside-Modified mRNA Influenza Virus Vaccine Provides Broad Protection in Mice. 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. 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. While most serologic assays are qualitative, a quantitative serologic . 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. The results resembled those observed in the panel that used a commercial recombinant S-trimer instead of transfected supernatant. By submitting a comment you agree to abide by our Terms and Community Guidelines. The function of secreted S protein also determined whether it could bind to hACE-2. van Doremalen, N. et al. 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. Some tests provide results rapidly (within minutes); others require 1-3 days for processing. Data are presented as GMT of micro-VNT50 titer with 95% confident interval. Another limitation was the lack of an external cohort to validate the suggested thresholds. 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. New crop of COVID-19 mRNA vaccines could be easier to store, cheaper to use: Science [updated 5 April 2022; cited 30 August 2022]. 7, eabi5246 (2021). Since COVID-19, the disease caused by severe acute respiratory virus 2 (SARS-CoV-2), began to spread in late December 2019, it has since become a global pandemic1. This is especially true of the mRNA vaccines, and the approach has shown better results than homologous prime-boost with a non-mRNA-based vaccine51. Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike. 007/2563), and the Armed Forces Research Institute of Medical Sciences, AFRIMS (IACUC approval no. For full functionality of this site, please enable JavaScript. N Engl J Med 384, 403416 (2021). In contrast, CoronaVac immunization showed the lowest T cells responses (42 SFC/106 splenocytes). At 2104 PFU of SARS-CoV-2 inoculum, PBS-vaccinated mice displayed clinical symptoms or weight loss within 1 day and all mice succumbed by day 6. 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. ChulaCov19 is therefore a promising mRNA vaccine candidate either as a primary or boost vaccination and has entered clinical development. Source data are provided as a Source Data file. Oran, D. P. & Topol, E. J. 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. Vaccines (Basel) 9, 874 (2021). This is consistent with a previous report46. Centrifuge RED TOP or EDTA tube and aliquot serum/plasma into plastic aliquot tube. Omicron spike function and neutralizing activity elicited by a comprehensive panel of vaccines. Ff!T8$I$I$I Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. By 18th August 2022, almost 600 million confirmed cases were caused by multiple VOCs and almost 6.5 million deaths were reported9. However, the slightly higher level compared to the 30-g group was not statistically significant. There were no anamnestic responses (four-fold increase on micro-VNT50 titers) in all vaccinated groups 6 days after the challenge, whereas one mouse in the control group developed a low micro-VNT50 titer at 40. a Kinetic response of micro-VNT50 titer after ChulaCov19 immunization and after challenge. Moreover, the feasibility of large-scale production as well as rapid adaptability to new variants are major advantages of the mRNA production platform. Testing for SARS-CoV-2 Infection. Background Identifying a specific threshold level of SARS-CoV-2 antibodies that confers protection in immunocompromised patients has been very challenging. Statistical analysis was performed using GraphPad Prism 9.0 software (San Diego, CA, USA). Its worth to mention that, as of now, theres no widely accepted cutoff value for immunity in immunocompromised patients, but some studies have suggested that antibody levels cut off may be associated with protection against COVID-19. https://ClinicalTrials.gov/show/NCT05231369 (2022). At 24h before transfection, 1105 Vero E6 cells were seeded in a 24-well plate (Thermo Fisher Scientific, MA, USA). Data Availability: All relevant data are within the manuscript and its Supporting Information files. Inclusion criteria were data from immunocompromised patients undergoing chemotherapy and/or biotherapy, aged over 18, who received at least one dose of the Pfizer/BioNTech BNT162b2 or Moderna COVID-19 vaccine (Spikevax) from three to six months before sampling collection. Cells with approximately 8090% confluency were transfected with 1g of IVT ChulaCov19 using Lipofectamine MessengerMax (Invitrogen, Carlsbad, CA, USA) according to the manufacturer protocols. Homologous prime/boost of each vaccine (CoronaVac, AZD1222, or ChulaCov19) were included as control groups. If testing will be delayed more than 7 days store at -20C or colder. The total volume of 50l of viral RNA was obtained from each sample. During the experiments, mice were maintained at 2022C and a relative humidity of 4510% on a 12h light/dark cycle. This observation correlates with that of a recent clinical study report53. A. et al. Alexander-Miller, M. A., Leggatt, G. R. & Berzofsky, J. In this study, the S1 and S2 subunits of the spike protein . World Health Organization. The Youden index indicates the performance (the larger the better) at a given cutoff: Youden = sensitivity + specificity 1 (the maximum value of the Youden index is 1) [17]. 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. In Experiment 3, the durability of NAb induced by ChulaCov19 was monitored until week 18 (15 weeks after the 2nd dose). 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. Increasing evidence suggests that vaccination regimens for COVID-19, that are applied to the general population, do not adequately protect a significant proportion of immunocompromised patients [5,6]. 4c). Post-translational modifications were also similar to those observed on SARS-CoV-241. Viral RNA was extracted from 140l serum and tissue samples using the QIAamp viral RNA mini kit (QIAGEN, Hilden, Germany). Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. S-specific total IgG analyzed at week 2 revealed that all ChulaCov19-immunized mice, either with 1 or 2 doses, elicited anti-S-specific IgG response from the lowest dose of 0.2g with a dose-dependent response pattern. 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. EBioMedicine 77, 103902 (2022). Zhang, L. et al. Bars represent the GMTs and 95% CI for each group. Common SARS-CoV-2 virus antigenic targets include spike, envelope, and nucleocapsid proteins [1]. The Abbott AdviseDx SARS-CoV-2 IgG II immunoassay detects antibodies to the viral spike protein (S). We recommend outside providers arrange to have their patients' blood drawn at their usual clinical draw sites and sent to the lab, preferably after contacting Client Support Services at commserv@uw.edu to facilitate testing. with these terms and conditions. BMC Med 20, 36 (2022). Jiang, R. D. et al. Magnitude of asymptomatic COVID-19 cases throughout the course of infection: A systematic review and meta-analysis. The female/male ratio was 67/33, and the median age was 47 years (IQR 3463). The opinions expressed here are the views of the writer and do not necessarily reflect the views and opinions of News Medical. Statistical significance was set at P < 0.05. Emerg Infect Dis 27, 31783180 (2021). These services aid in identifying a relative . The data supporting the findings of this work are available within the paper and in the Supplementary Information file. About the study. The most effective COVID-19 vaccines are mRNA-based and were first approved in the United Kingdom, the United States, and Europe. Agrawal, A. S. et al. Jackson, L. A. et al. The induced NAb was highly specific to the original variant, however, cross-neutralization against the VOCs was also observed. Frdrique Retornaz, CAS Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. Furthermore, the antibody rescue experiment confirmed the role of S1 in suppressing burst activities and highlighted the protective function of anti-S1 antibodies as well as the significance of RBD in modulating neuronal phenotypes. 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. https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. Medicines and Healthcare products Regulatory Agency (2022). From nanoparticle-based enrichment to mass spec refinements, they explore how these tools facilitate unbiased, deep, and rapid proteomics. In the clinical setting, >8 weeks interval for AZD1222 was recommended to maximize the vaccine efficacy52. So there is not enough data available to comment on the uptake of this therapy yet and raises the question in cases of previous infection or vaccination, the need to assess the SARS-CoV-2 antibody level for therapy decision making [1820]. Cohen J. 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. 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. Together with the emergence of new VOCs, a booster dose (either homologous or heterologous vaccine modality) is required to enhance the vaccine effectiveness15. The outcome strongly suggests that the RBD itself is sufficient to suppress surge activities. All authors reviewed the results and approved the final version of the manuscript. 3b). S protein on HEK293T-hACE-2 cell surface was stained with the same antibodies used in 2a. 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