These results confirmed that ChulaCov19 is highly immunogenic either as a primary vaccination in a vaccine-nave setting, or as a booster vaccine in animals previously vaccinated with other vaccines. It was subcloned into pUC-ccTEV-A101 using Afe I and Spe I restriction sites58. 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. Bars (a) or horizontal lines (b) represent the geometric mean (GMT) for each group while error bars indicate the 95% confident interval. Peer reviewer reports are available. Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen. A. Nature 608, 593602 (2022). 4c). Moreover, anamnestic NAb response is undetectable in challenge animals. By continuing to browse this site you agree to our use of cookies. DW, and MGA are named on patents that describe lipid nanoparticles for delivery of nucleic acid therapeutics, including mRNA and the use of modified mRNA in lipid nanoparticles as a vaccine platform. Please note that medical information found xA 0 @L This assay detects antibodies that block the interaction of SARS-CoV-2 with its entry receptor angiotensin-converting enzyme 2. CK was also funded by emerging Infectious Diseases and Vaccines Cluster, Ratchadapisek Sompoch Endowment Fund (2021), Chulalongkorn University (764002-HE04), and the Second Century Fund (C2F), Chulalongkorn University and Ratchadapiseksompotch Fund. 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. The remaining authors declare no competing interests. Cohen J. The promising preclinical study results presented here demonstrate that ChulaCov19 is highly immunogenic with protective efficacy. Experiment 2: heterologous prime-boost study, mice were primed with 1/10 of the approved human dosage of CoronaVac or AZD1222 and boosted 4 weeks later with 5g of ChulaCov19. Identifying a specific threshold level of SARS-CoV-2 antibodies that confers protection in immunocompromised patients has been very challenging. Hum Vaccin Immunother 12, 23512356 (2016). The neutralizing capacity was estimated by performing a surrogate virus neutralization test (sVNT) assay (GenScript, Piscataway, NJ, USA) as previously described [10,15,16]. Int J Infect Dis 112, 227234 (2021). In negative control (group 3), 5 mice were immunized with PBS instead of ChulaCov19 using the same schedule. 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. 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. Moreover, all five mice in control group exhibited varying symptoms of increased anorexia, lethargy, immobility, rough hair coat and increased respiration rate and effort. Article Developing highly effective vaccine platforms like mRNA technology in low- and middle-income countries (LMICs) is therefore an important goal21. 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. Nat Commun 14, 2309 (2023). A positive test result with the SARS -CoV-2 antibody test indicates that antibodies to SARS -CoV-2 were detected, and the individual has potential ly been exposed to Interim statement on the use of additional booster doses of Emergency Use Listed mRNA vaccines against COVID-19). 6c) may be due to RT-qPCR, a highly sensitive method detecting the free viral RNA from disintegrated virus. 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. Meanwhile, psVNT50 against BA.4/5 subvariant showed the lowest GMT in 1, 10, and 30g dosed groups. Solid reference line represents 264 binding antibody units (BAU)/ml cutoff (2.4 Log). Stphane Blachier, Splenocytes were collected at 2 weeks after the second dose (Experiment 1 & 2). It may also mean your body's immune system has generated a response to a prior COVID-19 infection. However, harmonization of neutralizing antibody titers is necessary to determine a common threshold using which vaccine protection can be predicted. {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^ 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. In all past pandemics, as well as the ongoing one with COVID-19, access to effective vaccines in a timely manner and has been severely limited in these countries. Zhang, N. N. et al. Vaccine inequity issue remains a major global challenge. How are the results reported for the anti-nucleocapsid antibody test, and what is the clinical significance? In response to the COVID19 pandemic and in preparation for future pandemics, Thailand has funded this mRNA vaccine development program from preclinical to manufacturing and clinical development. S-specific IFN- positive T cells were determined in duplicate assays from 5 mice in each group. Safety and immunogenicity of ChAdOx1 nCoV-19 vaccine administered in a prime-boost regimen in young and old adults (COV002): a single-blind, randomised, controlled, phase 2/3 trial. Nature Communications (Nat Commun) KR, DW, MGA, CK, EP, and SB are co-inventors of the submitted ChulaCov19 mRNA vaccines Patent. A subgroup analysis was performed according to vaccination status (complete or incomplete). 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]. Similar to the antibody results, the magnitude of T cell response was found to be dose-dependent but peaking at the 10-g dosage. "a97YEy111JlM7qqK;R]fr{g8 E]P7t iEx-m11tSmxsE,GE+hU#a=z1{/_vH}Nu&SENP_.*$ RL!DrojWs|[`}5C6nP,(n ,s-Km41vm8c/U3$@X3hUIwBge2Q{`4>4PQqo8"v3&v`wDXs%| 9>^8%|76sY6s$7PqI1QmO etbrr>$UmKd=UW-]Kd cg?q{`#*CM4\M6eKP2;:)U@(W$=u:{[9[S\2+wfynJ,%fd(~)qK5 The SARS-CoV-2 Spike IgG test shows the level of COVID-19 antibodies you had in your blood when you gave the blood sample. It was also evaluated for the protective efficacy in transgenic mice expressing human angiotensin-converting enzyme-2 (ACE2), Fig. 4a). Previous B cell depletion correlated with anti-SARS-CoV-2 IgG levels. mRNA capping was performed by the trinucleotide cap1 analog, CleanCap (TriLink Biotechnologies, San Diego, CA, USA). 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. The induced NAb was highly specific to the original variant, however, cross-neutralization against the VOCs was also observed. World Health Organization (2022). Overall, all assays showed good agreement with the Genscript sVNT. 4d). There are currently a few monoclonal antibody cocktails (such as bamlanivimab, casirivimab, and imdevimab together) that have been authorized by the US FDA for emergency use for the treatment of COVID-19 in certain population and similar medications have been authorized in other countries. 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. Liu, L. et al. World Health Organization. 399, 924944 (2022). : study conception and design, E.P., K.T., and C.K. Vaccines (Basel) 9, (2021). Tracking SARS-CoV-2 variants 2022 [updated 11 August 2022; cited 2022 19 August]. SARS-CoV-2 RNA-positive cells were examined and counted unblind by certified personnel. Research Use Only 2019-Novel Coronavirus (2019-nCoV) Real-time RT-PCR Primers and Probes) (2020). 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. The function of secreted S protein also determined whether it could bind to hACE-2. Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike. Virus titers were quantified by RT-qPCR and by determined the log10TCID50 values. Samples from 69 patients were analyzed. Cell nuclei were counter stained with 4, 6-diamino-2-phenylindole hydrochloride (DAPI) (Sigma-Aldrich, USA). 9, 1225 (2020). 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. Cao, Y. et al. Safety and immunogenicity of the ChAdOx1 nCoV-19 vaccine against SARS-CoV-2: a preliminary report of a phase 1/2, single-blind, randomised controlled trial. Therefore, the data indicated that the S1 subunit affected neurons only when the cells were exposed during the earliest stages of development. 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. In contrast, mice that received 2 doses of either 1 or 10 g of ChulaCov19 were normal with no symptoms throughout postchallenge period of 6 days. Google Scholar. The same dosage of approved vaccines were used with a dose of 5g ChulaCov19 (1/10 of the human dose used in Phase 2 Trial). The researchers believe that the study findings provide new insight into the activity of SARS-CoV-2 S proteins beyond their well-established functions in viral attachment and entry. Interestingly, the 3rd dose of ChulaCov19 administered at 17-week apart significantly boosted the NAb against all variants analyzed. The signal was amplified using a specific set of amplifiers (AMP1-6) as recommended by the manufacturer and was detected using a Fast Red solution for 5min at room temperature. Samples from 69 patients were included in this study. RNA copies were calculated as genomic equivalent/mg of tissue. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. For the homologous prime/boost, ChulaCov19 showed 3- to 10.6-fold higher NAb levels compared to 2-dose immunization of CoronaVac or AZD1222 across all variants WT (Wuhan-Hu1), Alpha (B.1.1.7), Beta B.1.351), and Delta (B.1.617.2), as measured by micro-VNT50 (Fig. PubMed 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. The average body weight by group from week 5 to week 5+6 days was demonstrated in Fig. J Control Release 217, 345351 (2015). KL and JH are employees of Genevant Sciences Corporation and are named on patent describing lipid nanoparticles. Agreements between antibody-binding assays and Genscript sVNT were performed using Cohens kappa, crude concordance rate, and area under curve (AUC). ChulaCov19 is therefore a promising mRNA vaccine candidate either as a primary or boost vaccination and has entered clinical development. The outcome strongly suggests that the RBD itself is sufficient to suppress surge activities. The OD450 of blanks were subtracted from OD450 of each sample before calculating antibody titer. 2a). 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. PubMed Central PN20-06). PubMed Sylvie Jordana, The data supporting the findings of this work are available within the paper and in the Supplementary Information file. Statistical significance was determined by two-sided MannWhitney test. World Health Organization. & Berzofsky, J. 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. The comparable molecular weight of S0 expressed by ChulaCov19 was also observed when using commercial recombinant S with S1/S2 cleavage site abolished as control (Fig. 11 Antibody tests may help identify past SARS-CoV-2 infection if performed two to four. 8 more], )5ul~eC}=,t?~]r6T5\OQhyN=8. These services aid in identifying a relative . Prompetchara, E. et al. Selective expansion of high- or low-avidity cytotoxic T lymphocytes and efficacy for adoptive immunotherapy. For example, the psVNT-50 against BA.1 in the CoronaVac-prime/ChulaCov19-boost group (psVNT-50 GMT=875) was significantly higher (p<0.01) than homologous CoronaVac (psVNT-50 GMT=5.1) and homologous AZD1222 (psVNT-50 GMT=2.7) groups. PLoS ONE 18(4): The bands corresponding to S1, S2 and intact S (S0) were detected. (2023, April 27). Article To detect SARS-CoV-2 RNA localization in mouse tissues samples, FFPE tissues of lung and nasal cavity were performed by using RNAscope In situ hybridization (ISH) assay. Safety and Immunogenicity of ChulaCov19 BNA159 mRNA Vaccine.). Per manufactures package insert protective level is 50.0 AU/mL. Labcorp offers both semi-quantitative and qualitative options for testing for spike protein antibodies. Buschmann, M. D. et al. Data Availability: All relevant data are within the manuscript and its Supporting Information files. Most convalescent patients tested with Tspot are reactive depending on which antigen is tested and which technique is used. All studies were conducted under protocols approved by the Committees on Care of Laboratory Animal Faculty of Medicine, Chulalongkorn University (IACUC approval no. Secreted S protein was also subjected for analysis of its binding capability to hACE2. Qualitative and semi-quantitative detection of antibodies to SARS-CoV-2 spike protein receptor binding domain (RBD). Vaccines (Basel) 10, 613 (2022). Homologous prime/boost of each vaccine (CoronaVac, AZD1222, or ChulaCov19) were included as control groups. The test can provide information about how your body reacted to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Four antibody binding assays were used for serological testing according to the instructions of the manufacturer. Emerg Infect Dis 27, 31783180 (2021). Google Scholar. This was followed by exposure to the same S1 concentration for seven consecutive days. Limited and Short-Lasting Virus Neutralizing Titers Induced by Inactivated SARS-CoV-2 Vaccine. 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. EBioMedicine 77, 103902 (2022). 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. The 4-week gap was used according to the preclinical study protocol of ChAdOX-vectored vaccines65,66. Some tests provide results rapidly (within minutes); others require 1-3 days for processing. 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. Among the 1g group, only one tissue had very few positive cells, the nasal epithelium. E.P., C.K., D.W., and K.R. Median time between last vaccination and sampling was 5.2 months (3.16.4). Data are presented as GMT of micro-VNT50 titer with 95% confident interval. For example, the micro-VNT50 GMT against WT (Wuhan-Hu1) in the AZD1222-prime/ChulaCov19-boost group was 7-fold higher than 2-dose AZD1222 immunization (GMT of micro-VNT50 were 31,042 vs 4457, p=0.0079). 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. This demonstrated the significant protective efficacy of ChulaCov19 in the preclinical phase. A Single-Cycle Influenza A Virus-Based SARS-CoV-2 Vaccine Elicits Potent Immune Responses in a Mouse Model. Eichinger, K. M. et al. The encapsulated mRNA-LNP was characterized by various parameters including size, polydispersity (PDI) and mRNA encapsulation efficiency at 1, 6, and 12 months after manufacture. The S protein facilitates virus attachment and entrance into the host cell. Nosoconseil, Aix les Bains, France, * E-mail: guillaume.penaranda@biogroup.fr. To address dose-response study of the ChulaCov19 and heterologous prime-boost responses with other approved COVID-19 vaccines, female BALB/c mice (Mus musculus), 4-6 weeks of age, (procured from Nomura Siam International, Bangkok, Thailand) were randomly divided into 5 mice/group for 3 sets of experiment. Note; the IgG2a/IgG1 ratio of 10g and 30g immunized mice were not analyzed due to limited volume of serum samples. More info. 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). The results resembled those observed in the panel that used a commercial recombinant S-trimer instead of transfected supernatant. 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. News-Medical, viewed 01 May 2023, https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. At 24h before transfection, 1105 Vero E6 cells were seeded in a 24-well plate (Thermo Fisher Scientific, MA, USA). Int J Infect Dis 114, 252260 (2022). In mice, ChulaCov19 was highly immunogenic as a booster in settings primed with either inactivated or viral vector vaccine. During the experiments, mice were maintained at 2022C and a relative humidity of 4510% on a 12h light/dark cycle. This is consistent with a previous report46. On day zero, neurons obtained from newborn P1 mice were treated with recombinant SARS-CoV-2 S protein and S1 and S2 S2 subunits. Roche Diagnostics, Inc. - Elecsys Anti-SARS-CoV-2 S. This test detects human SARS-CoV-2 antibodies . Retrieved on May 01, 2023 from https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. Centrifuge GOLD SST tube and route to Eastlake Virology (EVIR rack 81). PubMedGoogle Scholar. 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. Figures were created with BioRender.com. 4c). They were widely available in these countries for approximately a year before being accessible on other continents. All assays showed a high AUC for prediction of positive and negative results of Genscript sVNT (AUC > 0.90 for all) (Fig 2). The primary components of the SARS-CoV-2 structure are envelope (E), spike (S), membrane (M), and nucleocapsid (N) proteins. Laurent Kbaier, Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. This test should not be used to diagnose or exclude acute SARS-CoV-2 infection. Google Scholar. Bars represent the GMTs and 95% CI for each group. For western blot analysis, cell culture supernatant was analyzed by 12% polyacrylamide gel then transferred onto nitrocellulose membrane. Peletta, A. et al. PLOS ONE promises fair, rigorous peer review, Monoclonal anti-RBD (1:2,500), polyclonal-anti-S1 (1:5,000), -anti-S2 (1:5,000) or PSC (1:5,000) were used for detection of S protein in this step. Ferrara, F. & Temperton, N. Pseudotype Neutralization Assays: From Laboratory Bench to Data Analysis. 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. Freyn, A. W. et al. For the Siemens assay, the optimal cutoff was within the same range as the reference cutoff (270 BAU/ml).
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