Differences were considered significant at p<0.05 with exact p-values shown. 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). Each dot represents an individual animal. A Th2 dominant response following the vaccination remains a major concern of immunopathology that caused lung inflammation as reported in respiratory syncytial virus (RSV), SARS-CoV-1 and MERS-CoV42,43,44,45. Moreover, anamnestic NAb response is undetectable in challenge animals. Google Scholar. Nat Commun 13, 4610 (2022). In this interview conducted at Pittcon 2023 in Philadelphia, Pennsylvania, we spoke to Ron Heeran, a speaker at the 2023 James L. Waters Symposium. For SARS-CoV-2, tests to neutralize live viruses are performed only in specialized laboratories and are not standardized, making it difficult to compare and justify the use of a well-characterized sVNT as a functional reference [24,25].Additionally, neutralizing antibodies were not investigated, which could have helped in determining whether the anti-RBD or the anti-spike assays had the strongest correlation with virus neutralization. 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. 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. 4b). 200 0 obj <>]/Filter/FlateDecode/BitsPerComponent 8/Length 2211/Height 275>>stream Cell 182, 12711283.e1216 (2020). In each experiment, 3 internal controls (No Template Control (NTC), Negative Extraction Control (NEC) and Positive Extraction Control (PEC)) and 6 in vitro transcribed RNA standards were run along with test samples in each experiment. 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). Additional quality control to ensure the absence of double-stranded RNA (dsRNA) and endotoxin contamination prior to encapsulation into lipid nanoparticles (LNPs) were performed as described previously60. 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. Freyn, A. W. et al. Similar findings were also observed in BA.4/5 subvariant (Fig. Experiment 1: (a) Live-virus microneutralization (micro-VNT50) titers against WT (Wuhan-Hu1) live-virus at two weeks after receiving each vaccine dose. The vaccine was measured for its immunogenicity in BALB/c mice both using ChulaCov19 alone or as heterologous prime/boost regimens alongside the approved vaccines (Fig. 2a). ChAdOx1 nCoV-19 (AZD1222) or nCoV-19-Beta (AZD2816) protect Syrian hamsters against Beta Delta and Omicron variants. The neutralizing capacity was estimated by performing a surrogate virus neutralization test (sVNT) assay (GenScript, Piscataway, NJ, USA) as previously described [10,15,16]. Elecsys Anti-SARS-CoV-2 serology assay is intended for the detection of IgM and IgG antibodies to SARS-CoV-2 in human serum and plasma. All authors reviewed the results and approved the final version of the manuscript. 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. Stphane Blachier, Nature 589, 603607 (2021). Experiment 2: c micro-VNT50 titers against WT (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), and Delta (B.1.617.2) live-virus at two weeks after receiving various homologous or heterologous prime/boost regimens. By 18th August 2022, almost 600 million confirmed cases were caused by multiple VOCs and almost 6.5 million deaths were reported9. 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. 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. 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. 1a). Buschmann, M. D. et al. on this website is designed to support, not to replace the relationship The S1 subunit interacts with the angiotensin-converting enzyme 2 (ACE2) receptor present in the intestinal and lung cells. SD; standard deviation. Route to Eastlake Virology (EVIR rack 81). Vaccination status was complete among 61 patients (88%). 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. S-specific IgG measurement was performed employing indirect ELISA as described previously56,67. 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. 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]. Image Credit: whitehoune/Shutterstock.com. The total volume of 50l of viral RNA was obtained from each sample. New crop of COVID-19 mRNA vaccines could be easier to store, cheaper to use: Science [updated 5 April 2022; cited 30 August 2022]. This was followed by exposure to the same S1 concentration for seven consecutive days. The promising preclinical study results presented here demonstrate that ChulaCov19 is highly immunogenic with protective efficacy. Current SARS-CoV-2 antibody tests detect IgM or IgG to viral spike or nucleocapsid proteins. These results reflect that ChulaCov19 was highly immunogenic and induced a Th1-skewed response in mice. As with many viral respiratory infections, knowledge of the immune response to SARS-CoV-2 after a natural infection or vaccination, that could be predictive of the protection conferred, is challenging and not well established [14]. Mice were bled at 2 weeks after each dose and antibody responses were measured by ELISA and/or neutralization assays. In Experiment 3, the durability of NAb induced by ChulaCov19 was monitored until week 18 (15 weeks after the 2nd dose). PLoS One 16, e0249090 (2021). 8aU::fT23 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. One of these was the low number of samples that were subjected to antibody quantification and the absence of an independent international standard (WHO in IU/ml). Experiment 3: e psVNT50 NAb against WT (Wuhan-Hu1), Delta (B.1.617.2), and Omicron (BA.1 and BA.4/5) variants for NAb durability and effect of 3rd dose of ChulaCov19 studies. Am J Epidemiol 89, 422434 (1969). 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. 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. Science 377, 890894 (2022). These factors might make it difficult to draw a strong conclusion on vaccine efficacy from the current of experiments. In brief, 100ng of recombinant S-trimer (ACROBioSystems, China) were coated to the 96-well plates. Many types of tests are used to detect SARS-CoV-2, 1 and their performance characteristics vary. A SARS-CoV-2 probe (RNAscope Probe, V-nCoV2019-S, Advanced Cell Diagnostics ACD, Newark, CA (ACD, 848561)) was used. 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 . This neutralization antibody detection kit is designed to mimic the virus-host interaction utilizing recombinant RBD of the SARS-CoV-2 spike protein to detect antibodies that block the RBD binding to the hACE2 receptor. In the latter VNT protocol, serum-virus mixtures were incubated in VERO E6 cells for 5 days. Day 6 after the viral challenge (week 5+6 days), there was a slight decline of NAb titers in both groups but not statistically significant when compared to week 5, p=0.1126 and p=0.4437 for 10 g and 1 g groups, respectively. 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. Vaccines (Basel) 9, 850 (2021). Laurent Kbaier, Watanabe, Y. et al. In contrast, CoronaVac immunization showed the lowest T cells responses (42 SFC/106 splenocytes). For the Siemens assay, the optimal cutoff was within the same range as the reference cutoff (270 BAU/ml). Cell 185, 24222433.e2413 (2022). How are the results reported for the anti-nucleocapsid antibody test, and what is the clinical significance? analyse site usage and support us in providing free open access scientific content. 5a). Sera were collected at weeks 0, 2, 3, 4+6 days, and 5+6 days for NAb measurements. Hirabara, S. M. et al. Cell 182, 5058.e58 (2020). 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). Agreements between antibody-binding assays and Genscript sVNT were performed using Cohens kappa, crude concordance rate, and area under curve (AUC). Correspondence to PubMed Central The use of antibody therapy for PrEP, which is the use of medications to prevent infection before exposure to a virus, is currently being studied for its potential efficacy in immunocompromised individuals with COVID-19. In the nasal turbinate, vaccinated mice exhibited luminal accumulation of mucus and/or fibrin, albeit only minimal to mild amounts. This discovery may shed light on crucial aspects of SARS-CoV-2 infection, patient care methods, and future vaccine and antiviral development. Eichinger, K. M. et al. Testing for SARS-CoV-2 Infection. SARS-CoV-2 RNA-positive cells were examined and counted unblind by certified personnel. BMC Med 20, 36 (2022). Serologic Testing Serology testing measures the host antibody response in the form of immunoglobulins (Ig) such as IgM, IgA, or IgG following infection and/or vaccination. 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 . Study: SARS-CoV-2 Spike Protein Reduces Burst Activities in Neurons Measured by Micro-Electrode Arrays. The proprietary lipid and LNP composition are described in patent application WO2020097540A161,62. BA.2.12.1, BA.4 and BA.5 escape antibodies elicited by Omicron infection. 4d). Body weight of ChulaCov19 vaccinated mice decreased slightly only at days 1 and 2 post-challenge then gradually increased to the same levels as pre-challenge at day 6 (Fig. Kim, H. W. et al. Source data are provided as a source data file. Four antibody binding assays were used for serological testing according to the instructions of the manufacturer. There is also a limitation regarding the two semi-quantitative antibody binding assays as a saturation limit could be reached because of their limited measurement range. 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. Antibody escape of SARS-CoV-2 Omicron BA.4 and BA.5 from vaccine and BA.1 serum. Nosoconseil, Aix les Bains, France, * E-mail: guillaume.penaranda@biogroup.fr. NAb measurements in mice sera from Experiment 1 against WT (Wuhan-Hu1) live-virus (micro-VNT50) at 2-week after each dose showed NAb response in a dose-dependent manner. Statistical significance was determined by two-sided MannWhitney tests. Overall, all assays showed good agreement with the Genscript sVNT. In this study, ChulaCov19 was shown to be highly immunogenic, in a dose-responsive relationship, even when immunized with very low amount of 0.2g as measured by both live- and pseudovirus-neutralization assays. In the present study, researchers quantified the neurological phenotypes induced in neurons by the SARS-CoV-2 S protein. 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. It was also evaluated for the protective efficacy in transgenic mice expressing human angiotensin-converting enzyme-2 (ACE2), Fig. Data Availability: All relevant data are within the manuscript and its Supporting Information files. These tests should not be used to diagnosis or exclude acute SARS-CoV-2 infection. Samples from 69 patients were included in this study. 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. WW is an employee of BioNet-Asia, Co. Ltd. We have disclosed those interests fully to their affiliations, and we have in place an approved plan for managing any potential conflicts arising from licensing of the patents. Baden, L. R. et al. The ChulaCov19 vaccine development program has exactly this goal, striving to address the current and future pandemics in LMICs54. 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 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. However, there was no discernible difference in burst activity between S1-treated and the control wells. Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. Mice sera were further analyzed for NAb by psVNT50 test against the important recent VOCs, including Delta (B.1.617.2) variant and Omicron (BA.1 and BA.4/5) variants, and titers significantly decreased for all VOCs. Immunization with inactivated Middle East Respiratory Syndrome coronavirus vaccine leads to lung immunopathology on challenge with live virus. xA 0 @L Slides were counterstained with 50% Gill hematoxylin III (Sigma Aldrich, St Louis, MO, USA) for 2min and extensively washed under tap water. Zheng, C. et al. Agrawal, A. S. et al. RBD-VLP Vaccines Adjuvanted with Alum or SWE Protect K18-hACE2 Mice against SARS-CoV-2 VOC Challenge. During the experiments, mice were maintained at 2022C and a relative humidity of 4510% on a 12h light/dark cycle. Protective activity of mRNA vaccines against ancestral and variant SARS-CoV-2 strains. p<0.05 and p<0.01 are indicated by * and **, respectively.