: data collection, A.T., A.J., K.R., K.P., T.P., M.R., D.W., and K.R. Safety and Immunogenicity of Two RNA-Based Covid-19 Vaccine Candidates. Recommendations based on only one study is not prudent. News-Medical. : grant funding acquisition. 5a). Cannabis users with a genetic predisposition to schizophrenia more likely to experience psychotic symptoms. 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. Recombinant S protein with S1/S2 cleavage site abolished (ACROBioSystems, China) was used as positive control both in HEK293T-hACE-2 binding assay and western blot. After 2-dose, the GMTs of micro-VNT50 titer for 0.2, 1, 10, and 30g were 1280, 11,763, 54,047, and 62,084, respectively (Fig. The second dose of ChulaCov19 strongly augmented the IgG antibody levels with an increase of 10-19 folds, p<0.01 for all dose ranges (Fig. At week 5 (2 weeks after the second dose), all mice in both vaccinated groups showed increased NAb levels. BA.2.12.1, BA.4 and BA.5 escape antibodies elicited by Omicron infection. 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. In mice, ChulaCov19 was highly immunogenic as a booster in settings primed with either inactivated or viral vector vaccine. The presence of three SARS-CoV-2 genes (ORF1ab, nucleocapsid protein (N), and spike protein (S)) was identified using real-time PCR with the TaqPath RT-PCR COVID-19 kit (Thermo Fisher Scientific . 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. Overall, the rescue experiment provided compelling evidence that S1 was able to suppress burst activities when exposed to cells early in their developmental course. In conclusion, whether it is generally believed that a certain level of antibodies is necessary to confer protection against the virus, but the exact level required is not yet clear. The particles were re-characterized at 6- and 12-month after manufacture for stability assessment. Common SARS-CoV-2 virus antigenic targets include spike, envelope, and nucleocapsid proteins [1]. Since the outbreak of COVID-19, the world has raced to understand and accurately diagnose infection caused by SARS-CoV-2. As previously observed by Perkmann et al. Usually your antibody levels will go up after getting a vaccine or having an infection. Establishment of an mRNA vaccine platform in low- and middle-income countries (LMICs) is important to enhance vaccine accessibility and ensure future pandemic preparedness. 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. These authors jointly supervised this work: Drew Weissman, Kiat Ruxrungtham. It also markedly reduced viral RNA burden in serum and tissues. The authors acknowledge all the members of the Chula VRC for their input and support. SARS2Mutant: SARS-CoV-2 amino-acid mutation atlas database All samples were collected at the Alphabio Laboratory in Marseille, France (European Hospital, AlphabioBiogroup). The objective of the present study was to establish a new optimal threshold of protection for four different SARS-CoV-2 antibody assays [14]. James Heyes, A. J., Kieu Mong, L. A. M., Alan, D. MARTIN. 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. The micro-VNT50 titers was calculated as the reciprocal serum dilution that neutralized 50% of virus observed in virus control wells using probit analysis, SPSS program71. Previous B cell depletion correlated with anti-SARS-CoV-2 IgG levels. 2563.1/8 and 2564.1/4, National Research Council of Thailand NRCT. A recent randomized efficacy trial of the ChAdOx1 nCoV-19 (AZD1222) vaccine conducted in more than 8,500 patients in the United Kingdom, analyzed the antibody levels associated with protection against SARS-CoV-2 [7]. 8aU::fT23 The primary components of the SARS-CoV-2 structure are envelope (E), spike (S), membrane (M), and nucleocapsid (N) proteins. 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. Zhang, N. N. et al. Nanomaterial Delivery Systems for mRNA Vaccines. SARS-CoV-2 is an enveloped positive-sense single-stranded RNA beta coronavirus with a 30 kb polycistronic genome that encodes non-structural proteins (ORF1a and ORF1b, that are processed into Nsp1-16) at the 5-end, and structural proteins (S, E, M and N), and several other accessory factors (ORF3a . However, it has not been shown that COVID-19 mRNA vaccine encoding non-stabilized spike protein is not immunogenic or is not protective against viral challenge. . E.P., C.K., D.W., and K.R. PubMed Cellular and humoral responses after second and third SARS-CoV-2 Laboratoire BioestrelBiogroup, Mouans-Sartoux, France, Affiliation: The reactions were then stopped with 50L of 0.16N sulfuric acid. Mol Ther 28, 15691584 (2020). %PDF-1.7 3b). Google Scholar. The S protein trimer (S-trimer), depicted in Fig. In this interview conducted at Pittcon 2023 in Philadelphia, Pennsylvania, we spoke to Ron Heeran, a speaker at the 2023 James L. Waters Symposium. Secreted S protein was also subjected for analysis of its binding capability to hACE2. This assay detects antibodies that block the interaction of SARS-CoV-2 with its entry receptor angiotensin-converting enzyme 2. Another important limitation is that samples were collected at any time after the last vaccine dose (median 5.2 months (3.16.4)); Swadzba et al. In brief, 100ng of recombinant S-trimer (ACROBioSystems, China) were coated to the 96-well plates. 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). Statistical analysis was performed using GraphPad Prism 9.0 software (San Diego, CA, USA). wxWd~{Trru%m#97Z=}<8boK.3E@KT>1oW#!7q%7uJ?IC5 .iM!. In this interview conducted at Pittcon 2023 in Philadelphia, Pennsylvania, we spoke to Dr. Chad Merkin, Director of the International Institute for Nanotechnology, about his work developing next-generation nanomaterials for medical applications. Article The mid-point titers were determined in duplicate assays from 5 mice in each group. SARS-CoV-2 Antibody Profile, Nucleocapsid and Spike Efficacy and Safety of the mRNA-1273 SARS-CoV-2 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. Available from: https://www.bloomberg.com/graphics/covid-vaccine-tracker-global-distribution (2022). 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. Mid-point titers were calculated and expressed as the reciprocals of the dilution that showed an optical density (OD) at 50% of the maximum value substracted with the background (BSA plus secondary antibody). Source data are provided with this paper. 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. The purified mRNA-S (ChulaCov19) with undetectable endotoxin was tested for protein expression in VERO E6 cells. Source data are provided as a source data file. 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]. Splenocytes were collected at 2 weeks after the second dose (Experiment 1 & 2). Comparisons of the data between groups were made using non-parametric tests (MannWhitney test). The FFPE tissue slides were deparaffinized and treated with hydrogen peroxide (10min at room temperature) followed by target retrieval in 1X target retrieval solution in a steamer of at least 99C for 15min. The procedure of mouse IFN- ELISPOT used in this study was described in our previous reports56,72. Rotshild, V., Hirsh-Raccah, B., Miskin, I., Muszkat, M. & Matok, I. This candidate vaccine has now completed non-clinical toxicity and biodistribution studies and has entered Phase 1 and 2 human trials. All assays showed a high AUC for prediction of positive and negative results of Genscript sVNT (AUC > 0.90 for all) (Fig 2). 11 Antibody tests may help identify past SARS-CoV-2 infection if performed two to four. The opinions expressed here are the views of the writer and do not necessarily reflect the views and opinions of News Medical. Provided by the Springer Nature SharedIt content-sharing initiative. Vaccine inequity issue remains a major global challenge. Alene, M. et al. Notably, SARS-CoV-2 RNA measured by ISH was undetected in lung tissues in mice vaccinated with ChulaCov19 at either 1 or 10 g dose. Similar to the antibody results, the magnitude of T cell response was found to be dose-dependent but peaking at the 10-g dosage. Viruses were propagated in Vero E6 cells to generate sufficient titers 100TCID50 for the micro-VNT50 assay. 4e). ROC curves for each antibody binding assay according to Genscript sVNT. 3a). By 18th August 2022, almost 600 million confirmed cases were caused by multiple VOCs and almost 6.5 million deaths were reported9. Cohen J. 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. It also can show how your body reacted to COVID-19 vaccines. Stained cells were visualized under confocal microscope (ZEISS LSM 800, Carl Zeiss, Germany). However, there was no discernible difference in burst activity between S1-treated and the control wells. PLoS ONE 18(4): These results reflect that ChulaCov19 was highly immunogenic and induced a Th1-skewed response in mice. Correspondence to Nat Commun 13, 4610 (2022). Limited and Short-Lasting Virus Neutralizing Titers Induced by Inactivated SARS-CoV-2 Vaccine. 6b. Median time between last vaccination and sampling was 5.2 months (3.16.4). By clicking "Allow All" you agree to the storing of cookies on your device to enhance site navigation, 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. 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. Animals were immunized IM with 1g or 10g of ChulaCov19 at weeks 0 and 3. In summary, this mRNA vaccine development is an effort to set up the technology platform in LMICS. Laboratoires Oriade NovialeBiogroup, Grenoble, France, Affiliation: Beckman assay showed lower values as compared to all other assays (P< 0.008 for all paired comparisons); and lower values was observed for Siemens assay compared with Roche assay (P = 0.0033). An RBD virus-like particle vaccine for SARS-CoV-2 induces cross-variant 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. Cell nuclei were counter stained with 4, 6-diamino-2-phenylindole hydrochloride (DAPI) (Sigma-Aldrich, USA). Patrick Philibert, Kunkalikar, Bhavana. a Intracellular S protein expression examined by immunofluorescent assay employing anti-RBD, -S1, -S2 or PCS as primary antibody, the nuclei were counter stained with DAPI (blue). Statistical analysis was performed to compare the GMT of micro-VNT50 between 1 and 10 g dosed mice at each time point. For tissue samples, RNeasy Mini Kit (QIAGEN, Hilden, Germany) was used following manufacturer instructions.
