This would allow for identification of the corresponding thresholds, using high-throughput binding antibody assays. The particles were re-characterized at 6- and 12-month after manufacture for stability assessment. 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). It was subcloned into pUC-ccTEV-A101 using Afe I and Spe I restriction sites58. Two were quantitative: Abbott SARS-CoV-2 IgG II Quant-test (Abbott) (Abbott France, Rungis, France) with 50 arbitrary units (AU)/ml as a threshold for positivity, and Roche Elecsys anti-SARS-CoV-2 S (Roche Diagnostics France, Meylan, France) with 0.8 AU/ml used as a threshold for positivity. The results resembled those observed in the panel that used a commercial recombinant S-trimer instead of transfected supernatant. 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. Detection of antibodies to the SARS-CoV-2 spike glycoprotein in both serum and saliva enhances detection of infection LMICs received these vaccines much later and in shorter supply, as evidenced by the most recent statistic (as of 31 August 2022) that in several African countries less than 30% of the population has received at least one vaccine dose20. 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. Splenocytes from mice immunized with various dosages of ChulaCov19 (Experiment 1) were analyzed as summed frequency of S-specific IFN- positive T cells (Fig. 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. The authors would like to thanks Dr.Navapon Techakriengkrai, Faculty of Veterinary Science, Chulalongkorn University for providing HEK293T-hACE-2 cells. For a reference cutoff of 264 BAU/ml, assays showed moderate to good overall concordance with Genscript: 87% concordance for Abbott, 78% for Beckman, 75% for Roche, and 88% for Siemens. Biomedicines 10, 1464 (2022). The induced NAb was highly specific to the original variant, however, cross-neutralization against the VOCs was also observed. The team assessed the data using an algorithm devised in-house. World Health Organization. Please use one of the following formats to cite this article in your essay, paper or report: Kunkalikar, Bhavana. PubMed Central Citation: Halfon P, Jordana S, Blachier S, Cartlamy P, Kbaier L, Psomas CK, et al. : study conception and design, E.P., K.T., and C.K. Prolonged Protective Immunity Induced by Mild SARS-CoV-2 Infection of K18-hACE2 Mice. Viral RNA was extracted from 140l serum and tissue samples using the QIAamp viral RNA mini kit (QIAGEN, Hilden, Germany). SD; standard deviation. Please note that medical information found In the nasal turbinate, vaccinated mice exhibited luminal accumulation of mucus and/or fibrin, albeit only minimal to mild amounts. 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. a Kinetics of total IgG at 2 weeks after receiving 1 or 2 doses of 0.2, 1, 10, and 30g of ChulaCov19. In summary, this mRNA vaccine development is an effort to set up the technology platform in LMICS. 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. How are the results reported for the anti-nucleocapsid antibody test, and what is the clinical significance? T-cell responses to SARS-CoV-2 can be indirectly tested with antigen tests (such as Elispot) that tests for cytokines produced (i.e. Roche Diagnostics, Inc. - Elecsys Anti-SARS-CoV-2 S. This test detects human SARS-CoV-2 antibodies . About the study. The proprietary lipid and LNP composition are described in patent application WO2020097540A161,62. 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. ChAdOx1 nCoV-19 (AZD1222) or nCoV-19-Beta (AZD2816) protect Syrian hamsters against Beta Delta and Omicron variants. News-Medical. Ramasamy, M. N. et al. "Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons". Per manufactures package insert protective level is 50.0 AU/mL. Animals were immunized IM with 1g or 10g of ChulaCov19 at weeks 0 and 3. 1a). This study complied with the World Medical Association Declaration of Helsinki regarding the ethical conduct of research involving human subjects. 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. These viruses adapted to increase the transmissibility, severity and/or immune evasion8. S0 was used to depict unprocessed S protein. Cao, Y. et al. 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. 399, 924944 (2022). KR, DW, MGA, CK, EP, and SB are co-inventors of the submitted ChulaCov19 mRNA vaccines Patent. It is still being studied how does the immune system react in immunocompromised individuals, and how these observations translate into protection. Laboratoire AlphabioBiogroup, Marseille, France, Affiliation: Folegatti, P. M. et al. A positive result means your body's immune system has generated a response to the COVID-19 vaccine. Medicines and Healthcare products Regulatory Agency (2022). The LNP- encapsulated mRNA were characterized for their size, polydispersity using a Zetasizer (Zetasizer Nano DS, Malvern, UK), encapsulation efficiency, and shipped on dry ice and stored at 80 oC until use. Thank you for visiting nature.com. PLoS One 16, e0249090 (2021). Eichinger, K. M. et al. Samples from 69 patients were included in this study. Having more antibodies means your body can fight infection better than having fewer antibodies. 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. In contrast, CoronaVac immunization showed the lowest T cells responses (42 SFC/106 splenocytes). The S protein facilitates virus attachment and entrance into the host cell. 5a). ChulaCov19 is therefore a promising mRNA vaccine candidate either as a primary or boost vaccination and has entered clinical development. CAS The aim was to assess the threshold of 264 binding antibody units (BAU)/ml using four different SARS-CoV-2 antibody assays (Abbott, Beckman, Roche, and Siemens) and to establish a new optimal threshold of protection for each of the four . Vacharathit, V. et al. BA.2.12.1, BA.4 and BA.5 escape antibodies elicited by Omicron infection. (accessed May 01, 2023). At this time-point, the NAb titers against both Omicron subvariants were still in the same level with week 5 titers (Fig. All isolates were quantitated by tissue culture infectious dose TCID50 using the Reed-Muench method. Google Scholar. Previous specific optimal cutoffs fitted perfectly to patients with incomplete vaccination: a perfect agreement was observed between Genscript sVNT and each antibody binding assays among these patients (results not shown). Publishers note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. ChulaCov19 was further compared to two approved vaccines (CoronaVac and AZD1222), either in a homologous prime/boost setting or heterologous one (i.e. 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. At 2 weeks after the second immunization, mice were challenged intranasally with 2104 pfu (in 50L) of SARS-CoV-2 (wild-type). Oran, D. P. & Topol, E. J. 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. 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. In contrast, undetectable fluorescent signals for S proteins were observed when HEK293T-hACE-2 were incubated with supernatant from untransfected cells (Fig. Vaccines (Basel) 9, (2021). 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. Goat-anti-mouse IgG-FITC, donkey-anti-rabbit IgG-FITC (both were from BioLegend, CA, USA) or goat-anti-human AlexaFluor647 (Southern Biotech, AL, USA), at dilution of 1:5,000 was used as secondary antibodies following anti-RBD, -S1, -S2 or PCS staining. The results should always be assessed in conjunction with patient's medical history, clinical presentation, and other findings. 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. 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 . Within the brain, ACE2 is predominantly expressed in the brain stem and regions whose primary function is to regulate blood pressure and cardiovascular function. Interim statement on the use of additional booster doses of Emergency Use Listed mRNA vaccines against COVID-19). The team also determined whether the S1 subunit influences mature neurons during cell exposure. Interferon gamma) in response to SARS-CoV-2 antigens (M, N, S peptides). Liu, L. et al. [view The RNAscope ISH assay was performed using an RNAscope 2.5 HD Red Detection Kit (ACD, 322372) as followed. CAS As the Omicron subvariant BA.4/5 is currently spreading worldwide, we have also assessed cross-neutralization and found that the NAb GMT measured by psVNT50 against BA.4/5 in homologous ChulaCov19 vaccination or heterologous boosted with ChulaCov19 groups were significantly better than either of the CoronaVac or AZD1222 homologous vaccination (Fig. Lysis solution was added for 1h at RT before measuring OD at 540nm. 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. They concluded that higher levels of all immune markers were correlated with a reduced risk of symptomatic infection. Based on these studies, a threshold of 264 BAU/ml antibody was used as a recommendation for the use of PrEP in SARS-CoV-2 in France, and extrapolated to immunocompromised patients [9]. 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. Role of antigen, CD8, and cytotoxic T lymphocyte (CTL) avidity in high dose antigen induction of apoptosis of effector CTL. The mRNA vaccine technology transfer hub [updated 21 June 2022. 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. For full functionality of this site, please enable JavaScript. Translating a Thin-Film Rehydration Method to Microfluidics for the Preparation of a SARS-CoV-2 DNA Vaccine: When Manufacturing Method Matters. Samples from 69 patients were analyzed. 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. Nature 589, 603607 (2021). This study was performed using sera collected between October 2021 and December 2021 from a real life cohort of 69 individuals attending internal medicine and infectious diseases department of the European Hospital (Marseille). Moreover, the feasibility of large-scale production as well as rapid adaptability to new variants are major advantages of the mRNA production platform. on this website is designed to support, not to replace the relationship 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. The reaction was stopped by adding 50l/well of 0.5M sulfuric acid. Today, hundreds of commercial antibody tests are on the market despite often lacking proper validation and with unsatisfactory sensitivity and/or specificity. 4d). & Liu, J. Immunogenicity and safety of heterologous versus homologous prime-boost schedules with an adenoviral vectored and mRNA COVID-19 vaccine: a systematic review. Data are presented as GMT of micro-VNT50 titer with 95% confident interval. Recommendations based on only one study is not prudent. SARS-CoV-2 infection of human ACE2-transgenic mice causes severe lung inflammation and impaired function. 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. Safety and Immunogenicity of ChulaCov19 BNA159 mRNA Vaccine.). After 2 doses of ChulaCov19 or phosphate-buffered saline (PBS, control group) with a 3-week interval, K18-hACE2 mice were tested for NAb kinetics against live SARS-CoV-2 strain hCoV-19/Hongkong/VM20001061/2020. Article Source data are provided as a Source Data file. In addition, AZD1222 was also showed to be effective in clinical trials39,40. Moreover, ChAdOx1: AZD1222 that used unmodified S has been shown to induce high level of NAb and T cells responses even after a single immunization dose in two mouse strains38. 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. Pallesen, J. et al. Lancet. EBioMedicine 77, 103902 (2022). 2b). N Engl J Med 383, 24392450 (2020). CK, EP and KR were funded by the National Vaccine Institute (NVI), grant No. Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes. This demonstrated the significant protective efficacy of ChulaCov19 in the preclinical phase. Med. Of interest, the heterologous AZD1222-prime/ChulaCov19-boost induced the best specific T cells responses with mean spike-specific IFN- positive T cells of 3725 SFC/106 splenocytes, which approximately 1.7-fold higher than homologous ChulaCov19 (p=0.1934) and also significantly higher than other groups (p<0.05). Mice were bled at 2 weeks after each dose and antibody responses were measured by ELISA and/or neutralization assays. Note; 4 mice in 10g group were analyzed for psVNT50 against BA.4/5 due to the limited volume of serum samples. Wong, T. Y. et al. Agreement between antibody binding assays and Genscript sVNT positive and negative results according to optimal cutoff values determined for each assay. This was in accordance with previous studies showing that survived animals, either using HFH4-hACE2 or K18-hACE2 strains, could recovery their body weight to the basal levels at pre-challenge while weight lost continued for unprotected or non-vaccinated animals and reached euthanized criteria within approximately 1 week24,25,26. And the GMT NAb titer against WT (Wuhan-Hu1) in the CoronaVac-prime/ChulaCov19-boost group was also 7-fold higher than 2-dose of the CoronaVac group (GMT of micro-VNT50 were 23,525 vs 3378, p=0.0317), Fig. Pharmaceutics 14, 1427 (2022). BMC Med 20, 36 (2022). Quantitative data were reported using median and interquartile range (IQR), and qualitative data were reported using frequency and percentage. It is notable that while all mice, except for one, dosed with 10-g and 1-g ChulaCov19 showed no detectable SARS-CoV-2 viral RNA in tested tissues. Ying, B. et al. At week 5 (2 weeks after the second dose), all mice in both vaccinated groups showed increased NAb levels. 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. This elicited immunogenicity is significantly higher than those induced by homologous CoronaVac or AZD1222 vaccination. For group 1 and 2, there were 6 mice/group immunized intramuscularly via quadricep muscles with 2 doses, 3 weeks apart of ChulaCov19 at dose of 1g and 10g, respectively. Since the outbreak of COVID-19, the world has raced to understand and accurately diagnose infection caused by SARS-CoV-2. . Magnitude of asymptomatic COVID-19 cases throughout the course of infection: A systematic review and meta-analysis. Thus, most recently approved vaccines employ full-length S (with or without modification) or whole virus (inactivated) as a target antigen19. 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. There were few limitations in this study. The most effective COVID-19 vaccines are mRNA-based and were first approved in the United Kingdom, the United States, and Europe. After a mean (SD) of 19 (10) and 16 (2) days from the second and third vaccine dose, IgG anti-SARS-CoV-2 antibodies were detected in 6 (60%) and 8 (80%) patients, respectively. Protection against COVID-19 is thought to depend on the presence of specific antibodies against the virus, as well as the function of other components of the immune system such as T cells. 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). All assays showed a high AUC for prediction of positive and negative results of Genscript sVNT (AUC > 0.90 for all) (Fig 2). WIPO (2020). To test the hypothesis that the S1 receptor-binding domain (RBD) may be the reason for burst reduction, the team collected and assessed purified recombinant RBD. Briefly, 100ng/well of RBD recombinant proteins (Abcam, UK) were coated overnight to the 96-well plates. Julie Allemand-Sourrieu, The plasmid was propagated in E. coli (Stbl3, Invitrogen, Carlsbad, CA, USA) and extracted by EndoFree Giga Kit (Qiagen, Hilden, Germany). van Doremalen, N. et al. Bloomberg. No positive detection of viral RNA was present in the 10g group of animals analyzed by ISH. Prompetchara, E., Ketloy, C., Alameh, MG. et al. Solid reference line represents 264 binding antibody units (BAU)/ml cutoff (2.4 Log). Nucleoside-modified mRNA was produced by in vitro transcription (IVT) by substitution of uridine triphosphate (UTP) with N1-methylpseudouridine (m1) triphosphate (TriLink, Biotechnologies, San Diego, CA, USA), detailed elsewhere58. IVT mRNA was analyzed on agarose for determination of its integrity. Nature 586, 578582 (2020). In the homologous prime/boost of these 2 approved vaccines groups, each was given at four weeks interval. Immunogenicity and protective efficacy of SARS-CoV-2 mRNA vaccine encoding secreted non-stabilized spike in female mice, https://doi.org/10.1038/s41467-023-37795-0. Sci Transl Med 14, eabm3302 (2022). A. 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. The 4-week gap was used according to the preclinical study protocol of ChAdOX-vectored vaccines65,66. 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. Meanwhile, psVNT50 against BA.4/5 subvariant showed the lowest GMT in 1, 10, and 30g dosed groups. 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]. 2023. In the control group, positive viral RNA staining was present in individual neurons of the olfactory bulb (4/4), epithelial cells of the nasal sinus (4/5), alveolar epithelial cells and macrophages in the lung (5/5), see Table1. This test should not be used to diagnose or exclude acute SARS-CoV-2 infection. 3b). For western blot analysis, cell culture supernatant was analyzed by 12% polyacrylamide gel then transferred onto nitrocellulose membrane. The images or other third party material in this article are included in the articles Creative Commons license, unless indicated otherwise in a credit line to the material. Interestingly, the 3rd dose of ChulaCov19 administered at 17-week apart significantly boosted the NAb against all variants analyzed. Tracking SARS-CoV-2 variants 2022 [updated 11 August 2022; cited 2022 19 August]. UW MedicineDepartment of Laboratory MedicineVirology- Covid Testing Lab1601 Lind Ave SWRenton, WA 980573356Tel: (206) 685-6656 opt 4. On the basis of these data at present anti-SARS CoV-2 serological assays' results are not interchangeable, and, more importantly, individual immune monitoring should be performed with the same method.
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