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. 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). The differences in the commercial assays used in this study are related to the components of the tests (the spike antigen epitopes used, the different isolates of the SARS-CoV-2, and the quantification of either total antibodies or only IgG) [2123]. 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 . ];V^srE]DwCyTPre_fyG;Cb@*\d$ j '-|,!]jF/J9r\s%3! At week 18, the NAb against WT (Wuhan-Hu1) and Delta (B.1.617.2) decreased approximately 2-fold but not statistically significant when compare with week 5 titers. 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. In this study, the S1 and S2 subunits of the spike protein were evaluated separately to determine if they elicited any neurological phenotypes as estimated by the micro-electrode arrays (MEAs). In contrast, CoronaVac immunization showed the lowest T cells responses (42 SFC/106 splenocytes). Agreements between antibody-binding assays and Genscript sVNT were performed using Cohens kappa, crude concordance rate, and area under curve (AUC). 199 0 obj <>stream the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in The overall concordance between the antibody binding assays and the Genscript sVNT also increased consistently i.e., 11% increase for Roche (86% concordance), 10% increase for Beckman (88% concordance), 2% increase for Siemens (90% concordance), and 1% increase for the Abbott assay (88% concordance). For western blot analysis, cell culture supernatant was analyzed by 12% polyacrylamide gel then transferred onto nitrocellulose membrane. Available from: https://www.science.org/content/article/new-crop-covid-19-mrna-vaccines-could-be-easier-store-cheaper-use (2022). Note: tissues from 3/5 animals in control group were collected at day 5. Statistical analysis significance was determined by two-sided MannWhitney test. 5a). Article After SARS-CoV-2 challenge, there was no measurable decline in body weight among vaccinated groups. SARS-CoV-2 neutralizing antibodies decline over one year and patients with severe COVID-19 pneumonia display a unique cytokine profile. Developing mRNA vaccine technology for distribution in these regions is therefore extremely important21. On the contrary, low avidity T cells which require a higher amount of viral antigen were able to lyse the viral infection after the new virion were produced31. Omicron spike function and neutralizing activity elicited by a comprehensive panel of vaccines. Results were determined as a ratio of the signal of the samples to the average signal of calibrators. Bars represent the GMTs and 95% CI for each group. Broad and timely access to effective vaccines in LMICs, particularly the most under-served settings, has always been limited during past pandemics and this has extended to COVID-1920. The 5-fold serially diluted mice sera were added in duplicate. 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. SARS-CoV-2 RNA-positive cells were examined and counted unblind by certified personnel. xA 0 @L 4b). Kunkalikar, Bhavana. In this episode of omg OMx, Bruker's science-driven podcast, Kate Stumpo interviews Daniel Hornburg, the VP of Proteomics at Seer, as they discuss the innovative technologies in plasma proteomics. Correspondence to ADS Differences were considered significant at p<0.05 with exact p-values shown. 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. endstream Splenocytes were collected at 2 weeks after the last dose (Experiment 1 & 2) for assessment of spike-specific IFN- T-cell using ELISpot assay (Fig. Magnitude of asymptomatic COVID-19 cases throughout the course of infection: A systematic review and meta-analysis. 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. 6a). 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]. 1a). & Berzofsky, J. 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. 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 . Mean spike-specific IFN- positive T cells for 0.2, 1, 10 and 30g were 166, 429, 1913, and 1378 SFC/106 splenocytes, respectively. Here, we describe the preclinical studies of ChulaCov19, a SARS-CoV-2 mRNA encoding prefusion-unstabilized ectodomain spike protein encapsulated in lipid nanoparticles (LNP). 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. Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. In the homologous prime/boost of these 2 approved vaccines groups, each was given at four weeks interval. 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. What are the benefits of exercise on cardiovascular health. The use of a specific threshold for decision-making regarding PreP in immunocompromised patients must be taken with cautions due to limitations, mainly based on the nature and type of assay used to measure the antibody. In the latter VNT protocol, serum-virus mixtures were incubated in VERO E6 cells for 5 days. Hum Vaccin Immunother 12, 23512356 (2016). Comparisons of the data between groups were made using non-parametric tests (MannWhitney test). When RT-qPCR was used, although viral RNA was still detected in some tissues, both dosages demonstrated a 99-100% reduction of viral RNA in tested tissues when compared to the control group. sample is stable for 7 days at 2-8C once separated from a clot or red blood cells, or in a gel separator tube. 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. 4d). https://apps.who.int/iris/handle/10665/363344 (2022). Two were semi-quantitative: Beckman Access SARS-CoV-2 IgG II (Beckman Coulter France SAS, Roissy CDG, France) with 30 AU/ml as a threshold for positivity and Siemens Atellica IM SARS-CoV-2 IgG (Siemens Healthcare SAS, Saint-Denis, France) with 0.8 AU/ml used as a threshold for positivity. The Wilcoxon test for pairwise comparisons yielded P < 0.0001 for all comparisons. analyse site usage and support us in providing free open access scientific content. Previous studies reported that low-dose vaccination induced only high avidity T cells. Labcorp test details for SARS-CoV-2 Semi-Quantitative Total Antibody, Spike . Samples from 69 patients were analyzed. Another limitation was the lack of an external cohort to validate the suggested thresholds. The most effective COVID-19 vaccines are mRNA-based and were first approved in the United Kingdom, the United States, and Europe. 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. FITC-tagged 2nd Abs (green) were used for detection of RBD, S1, and S2 while AlexaFluor647-tagged 2nd Ab (red) was used following PCS staining. N Engl J Med 384, 403416 (2021). The GMT of micro-VNT50 titers at week 5 were 15,343 and 4424 in the 10 g and 1 g groups, respectively, p=0.0325. The nonparametric KruskalWallis test for multiple comparisons was used to compare all assays. Nature 586, 578582 (2020). Qualitative and semi-quantitative detection of antibodies to SARS-CoV-2 spike protein receptor binding domain (RBD). All data were fully anonymized before the analysis. Jairak, W. et al. An mRNA Vaccine against SARS-CoV-2 - Preliminary Report. In addition, there was no anamnestic antibody response detected in the ChulaCov19 vaccinated mice after viral challenge (Fig. By continuing to browse this site you agree to our use of cookies. Each dot represents an individual animal. 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. Calculations were performed using the SAS V9.4 software (SAS Institute Inc., Cary, NC, USA). This study also indicated that neutralizing S1 restores neuronal discharge activities to control levels. It also can show how your body reacted to COVID-19 vaccines. In the lung, inflammation was limited to predominantly peribronchiolar proliferation of mononuclear cells, akin to an expansion of cellularity among bronchiolar lymphoid tissue but without notable follicle formation. Klemis, V. et al. Covid-19 Tracker: More Than 12.6 Billion Shots Given [updated 31 August 2022; cited 2022 31 August]. Source data are provided as a source data file. Front Cell Infect Microbiol 11, 781429 (2021). 6c) may be due to RT-qPCR, a highly sensitive method detecting the free viral RNA from disintegrated virus. Source data are provided as a Source Data file. These results reflect that ChulaCov19 was highly immunogenic and induced a Th1-skewed response in mice. 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. Comparable to the S1 data, the team identified a significant reduction in surge activities. This discovery may shed light on crucial aspects of SARS-CoV-2 infection, patient care methods, and future vaccine and antiviral development. "Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons". Viruses were propagated in Vero E6 cells to generate sufficient titers 100TCID50 for the micro-VNT50 assay. 8aU::fT23 : draft manuscript preparation. On Day 5, significant weight reduction (p<0.05) was observed in control group when compared with the vaccinated groups. It is still being studied how does the immune system react in immunocompromised individuals, and how these observations translate into protection. p<0.05 and p<0.01 are indicated by * and **, respectively. Stability: Sample stable off the clot, red blood cells, or separator gel for 7 days at 2-8C. Viral RNA was extracted from 140l serum and tissue samples using the QIAamp viral RNA mini kit (QIAGEN, Hilden, Germany). PubMed Central The slides were dehydrated in 60C dry oven until completely dry and then dipped in Xylene before mounting with a mounting medium. The S1 subunit interacts with the angiotensin-converting enzyme 2 (ACE2) receptor present in the intestinal and lung cells. These tests should not be used to diagnosis or exclude acute SARS-CoV-2 infection. Therefore, the data indicated that the S1 subunit affected neurons only when the cells were exposed during the earliest stages of development. A. PubMed Central p<0.05 and p<0.01 are indicated by * and **, respectively. Therefore SARS-CoV-2 serology may be standardized. Jiang, R. D. et al. In addition, AZD1222 was also showed to be effective in clinical trials39,40. 9, 1225 (2020). Figures were created with BioRender.com. Philippe Halfon, Immunization with inactivated Middle East Respiratory Syndrome coronavirus vaccine leads to lung immunopathology on challenge with live virus. 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. The average body weight by group from week 5 to week 5+6 days was demonstrated in Fig. After 1h incubation at 37C, plates were washed vigorously with washing buffer (PBS+0.5% Tween 20, PBST). 399, 924944 (2022). 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]. 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. {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 negative control (group 3), 5 mice were immunized with PBS instead of ChulaCov19 using the same schedule. 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. b hACE-2 binding assay (merged): culture supernatant collected from ChulaCov19 transfected cells incubated with HEK293T- hACE-2 cells. Stphane Blachier, c S protein expression in cell culture supernatant analyzed by western blot using anti-RBD, -S1, -S2 or PCS as primary antibody. Frdrique Retornaz, Zhang, N. N. et al. ADS This was consistent with the prior study in K18-hACE2 that intranasal inoculation with the similar range of virus caused death within 1 week22. World Health Organization. 4c). 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. Vaccines (Basel) 9, 850 (2021). Laboratoire AlphabioBiogroup, Marseille, France, 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]. Statistical analysis was performed to compare the GMT of micro-VNT50 between 1 and 10 g dosed mice at each time point. After the first dose, NAb were detected in mice that received 1, 10, and 30g ChulaCov19 with corresponding GMTs of micro-VNT50 titer of 80, 368, and 735, respectively. Using western blot, the S protein could be detected in cell culture supernatant when using anti-RBD, -S1, -S2 and PCS as primary antibodies. It was also evaluated for the protective efficacy in transgenic mice expressing human angiotensin-converting enzyme-2 (ACE2), Fig. 5b). \1;nJ/mjJ=DqXlU,u>z}x)tU#K>/#}idN"%W$YoSA14Ys5+VlE5-3a+`h"xD%5n#L$\g%[&0Gy-x;a>$'+6#am#WK>nxW|^E~YS t4G2G9V$Mf=E5y? showed time-dependent changes in the comparability of different antibody tests with samples collected at different time points [26]. All studies were conducted under protocols approved by the Committees on Care of Laboratory Animal Faculty of Medicine, Chulalongkorn University (IACUC approval no. CAS The study suggested that S1 is responsible for decreasing burst activities of neuronal populations when cells are exposed early in the course of development. For patients who do not regularly seek care within UW Medicine, our phlebotomists at the University of Washington Medical Center-Northwest Campus (UWMC-NW) and UWMC-NW Outpatient Medical Center (OPMC) located on Meridian Ave. N. are able to perform blood draws for testing with a valid provider order. CAS Tissues were collected at week 5+6 days for assessment of viral RNA. The function of secreted S protein also determined whether it could bind to hACE-2. Google Scholar. 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 same dosage of approved vaccines were used with a dose of 5g ChulaCov19 (1/10 of the human dose used in Phase 2 Trial). S-specific IFN- positive T cells were determined in duplicate assays from 5 mice in each group. 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. SARS-CoV-2, SARS-CoV, and MERS-CoV viral load dynamics, duration of viral shedding, and infectiousness: a systematic review and meta-analysis. RBD-VLP Vaccines Adjuvanted with Alum or SWE Protect K18-hACE2 Mice against SARS-CoV-2 VOC Challenge. Voysey, M. et al. https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. Indeed, antibody therapy for pre-exposure prophylaxis (PrEP), may be efficient in preventing hospitalization in immunocompromised patients, regardless of the variant involved. 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. ~N6R}q71m*:H^J6n1i2bcPh)\R ^Zei%|kz1>85k7^T\Lt)h\=-an}mr^yU+HNGlcE'&ps4toE4/f_=3A>o'9p:4ZUI1s+QtSIwu,\\Uv+QG Pm2+T`2T\y/3^?8UlVshDEY/qo_F%3^>D?lq2g R61sbWP}O3,p?K|3Tx=RdR'_/KzVFS j\h1s1vdzV|:p Christina K. Psomas, x1 AQ1DIt-~+qc U/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J R/ )@J _3 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. 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.