scholarly journals Towards Internationally standardised humoral Immune Correlates of Protection from SARS CoV 2 infection and COVID-19 disease

2021 ◽  
Author(s):  
Javier Castillo-Olivares ◽  
David Wells ◽  
Matteo Ferrari ◽  
Andrew Chan ◽  
Peter Smith ◽  
...  
Keyword(s):  
Large Scale ◽  
Antibody Responses ◽  
Clinical Severity ◽  
World Health ◽  
Binding Assays ◽  
Neutralising Antibodies ◽  
Correlates Of Protection ◽  
Immune Correlates ◽  
Antibody Assays ◽  
Binding Antibody

Precision monitoring of antibody responses during the COVID-19 pandemic is increasingly important during large scale vaccine rollout and rise in prevalence of Severe Acute Respiratory Syndrome- related Coronavirus-2 (SARS-CoV-2) variants of concern (VOC). Equally important is defining Correlates of Protection (CoP) for SARS-CoV-2 infection and COVID-19 disease. Data from epidemiological studies and vaccine trials identified virus neutralising antibodies (VNab) and SARS- CoV-2 antigen-specific (notably RBD, and S) binding antibodies as candidate CoP. In this study, we used the World Health Organisation (WHO) international standard to benchmark neutralising antibody responses and a large panel of binding antibody assays to compare convalescent sera obtained from: a) COVID-19 patients; b) SARS-CoV-2 seropositive healthcare workers (HCW) and c) seronegative HCW. The ultimate aim of this study, was to identify biomarkers of humoral immunity that could be used as candidate CoP in internationally accepted unitage. Whenever suitable, the antibody levels of the samples studied were expressed in International Units (INU) for virus neutralisation assays or International Binding Antibody Units (BAU) for ELISA tests. In this work we used commercial and non-commercial antibody binding assays; a lateral flow test for detection of SARS-CoV-2-specific IgG / IgM; a high throughput multiplexed particle flow cytometry assay for SARS-CoV-2 Spike (S), Nucleocapsid (N) and Receptor Binding Domain (RBD) proteins); a multiplex antigen semi-automated immuno-blotting assay measuring IgM, IgA and IgG; a pseudotyped microneutralisation test (pMN) and electroporation-dependent neutralisation assay (EDNA). Our results indicate that overall, severe COVID-19 patients showed statistically significantly higher levels of SARS-CoV-2-specific neutralising antibodies (average 1029 IU/ml) than those observed in seropositive HCW with mild or asymptomatic infections (379 IU/ml) and that clinical severity scoring, based on WHO guidelines was tightly correlated with neutralisation and RBD / S binding assays. In addition, there was a positive correlation between severity, N-antibody assays and intracellular virus neutralisation.

scholarly journals Analysis of Serological Biomarkers of SARS-CoV-2 Infection in Convalescent Samples From Severe, Moderate and Mild COVID-19 Cases

2021 ◽  
Vol 12 ◽  
Author(s):  
Javier Castillo-Olivares ◽  
David A. Wells ◽  
Matteo Ferrari ◽  
Andrew C. Y. Chan ◽  
Peter Smith ◽  
...  
Keyword(s):  
Large Scale ◽  
World Health Organisation ◽  
Epidemiological Studies ◽  
Antibody Responses ◽  
Clinical Severity ◽  
World Health ◽  
Binding Assays ◽  
Neutralising Antibodies ◽  
Antibody Assays ◽  
Binding Antibody

Precision monitoring of antibody responses during the COVID-19 pandemic is increasingly important during large scale vaccine rollout and rise in prevalence of Severe Acute Respiratory Syndrome-related Coronavirus-2 (SARS-CoV-2) variants of concern (VOC). Equally important is defining Correlates of Protection (CoP) for SARS-CoV-2 infection and COVID-19 disease. Data from epidemiological studies and vaccine trials identified virus neutralising antibodies (Nab) and SARS-CoV-2 antigen-specific (notably RBD and S) binding antibodies as candidate CoP. In this study, we used the World Health Organisation (WHO) international standard to benchmark neutralising antibody responses and a large panel of binding antibody assays to compare convalescent sera obtained from: a) COVID-19 patients; b) SARS-CoV-2 seropositive healthcare workers (HCW) and c) seronegative HCW. The ultimate aim of this study is to identify biomarkers of humoral immunity that could be used to differentiate severe from mild or asymptomatic SARS-CoV-2 infections. Some of these biomarkers could be used to define CoP in further serological studies using samples from vaccination breakthrough and/or re-infection cases. Whenever suitable, the antibody levels of the samples studied were expressed in International Units (IU) for virus neutralisation assays or in Binding Antibody Units (BAU) for ELISA tests. In this work we used commercial and non-commercial antibody binding assays; a lateral flow test for detection of SARS-CoV-2-specific IgG/IgM; a high throughput multiplexed particle flow cytometry assay for SARS-CoV-2 Spike (S), Nucleocapsid (N) and Receptor Binding Domain (RBD) proteins); a multiplex antigen semi-automated immuno-blotting assay measuring IgM, IgA and IgG; a pseudotyped microneutralisation test (pMN) and an electroporation-dependent neutralisation assay (EDNA). Our results indicate that overall, severe COVID-19 patients showed statistically significantly higher levels of SARS-CoV-2-specific neutralising antibodies (average 1029 IU/ml) than those observed in seropositive HCW with mild or asymptomatic infections (379 IU/ml) and that clinical severity scoring, based on WHO guidelines was tightly correlated with neutralisation and RBD/S antibodies. In addition, there was a positive correlation between severity, N-antibody assays and intracellular virus neutralisation.

scholarly journals SARS-CoV-2 antibody binding and neutralization in dried blood spot eluates and paired plasma

2021 ◽  
Author(s):  
Hannah L. Itell ◽  
Haidyn Weight ◽  
Carolyn S. Fish ◽  
Jennifer K. Logue ◽  
Nicholas Franko ◽  
...  
Keyword(s):  
Dried Blood Spots ◽  
Antibody Binding ◽  
Antibody Responses ◽  
Cold Chain ◽  
Correlates Of Protection ◽  
Antibody Assays ◽  
Plasma Antibody ◽  
Highly Correlated ◽  
And Storage ◽  
Blood Spot

Widescale assessment of SARS-CoV-2-specific antibodies is critical to understanding population seroprevalence, correlates of protection, and the longevity of vaccine-elicited responses. Most SARS-CoV-2 studies characterize antibody responses in plasma/sera. While reliable and broadly used, these samples pose several logistical restrictions such as requiring venipuncture for collection and cold chain for transportation and storage. Dried blood spots (DBS) overcome these barriers as they can be self-collected by fingerstick and mailed and stored at ambient temperature. Here, we evaluate the suitability of DBS for SARS-CoV-2 antibody assays by comparing several antibody responses between paired plasma and DBS from SARS-CoV-2 convalescent and vaccinated individuals. We found that DBS not only reflected plasma antibody binding by ELISA and epitope profiles using phage-display, but also yielded SARS-CoV-2 neutralization titers that highly correlated with paired plasma. Neutralization measurement was further streamlined by adapting assays to a high-throughput 384-well format. This study supports the adoption of DBS for numerous SARS-CoV-2 binding and neutralization assays.

scholarly journals Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples

2017 ◽  
Author(s):  
Thomas Liechti ◽  
Claus Kadelka ◽  
Hanna Ebner ◽  
Nikolas Friedrich ◽  
Roger D. Kouyos ◽  
...  
Keyword(s):  
High Throughput ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Assay System ◽  
Clinical Samples ◽  
Correlates Of Protection ◽  
Technical Advances ◽  
Broadly Neutralizing Antibody ◽  
Binding Antibody ◽  
Hiv 1

AbstractThe monitoring and assessment of a broadly neutralizing antibody (bnAb) based HIV-1 vaccine require detailed measurements of HIV-1 binding antibody responses to support the detection of correlates of protection. Here we describe the development of a flexible, high-throughput microsphere based multiplex assay system that allows monitoring complex binding antibody signatures. Studying a panel of 13 HIV-1 antigens in a parallel assessment of different IgG subclasses (IgG1, IgG2 and IgG3) we demonstrate the potential of our strategy. The technical advances we describe include means to improve antigen reactivity using directed neutravidin-biotin immobilization of antigens and biotin saturation to reduce background. A particular emphasis of our study was to provide tools for the assessment of reproducibility and stability of the assay system and strategies to control for variations allowing the application in high-throughput assays, where reliability of single measurements needs to be guaranteed.

scholarly journals Immunological surrogate endpoints of COVID-2019 vaccines: the evidence we have versus the evidence we need

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Pengfei Jin ◽  
Jingxin Li ◽  
Hongxing Pan ◽  
Yanfei Wu ◽  
Fengcai Zhu
Keyword(s):  
Large Scale ◽  
Current Knowledge ◽  
Surrogate Endpoints ◽  
Phase 3 ◽  
Vaccine Candidates ◽  
Correlates Of Protection ◽  
Immune Correlates ◽  
Efficacy Trials ◽  
Review Current

AbstractIn response to the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic, over 200 vaccine candidates against coronavirus disease 2019 (COVID-2019) are under development and currently moving forward at an unparalleled speed. The availability of surrogate endpoints would help to avoid large-scale filed efficacy trials and facilitate the approval of vaccine candidates, which is crucial to control COVID-19 pandemic. Several phase 3 efficacy trials of COVID-19 vaccine candidates are under way, which provide opportunities for the determination of COVID-19 correlates of protection. In this paper, we review current knowledge for existence of COVID-19 correlates of protection, methods for assessment of immune correlates of protection and issues related to COVID-19 correlates of protection.

scholarly journals Human antibody responses following vaccinia immunization using protein microarrays and correlation with cell-mediated and antibody dependent cellular cytotoxicity responses

2021 ◽  
Author(s):  
Sharon E Frey ◽  
Jack T Stapleton ◽  
Zuhair K Ballas ◽  
Wendy L Rasmussen ◽  
Thomas M Kaufman ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Protein Microarray ◽  
Protein Microarrays ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Antibody ◽  
Cellular Cytotoxicity ◽  
Antibody Dependent Cellular Cytotoxicity ◽  
Binding Assays ◽  
Correlates Of Protection

Abstract Background There are limited data regarding immunological correlates of protection for the modified vaccinia Ankara (MVA) smallpox vaccine. Methods Five hundred twenty-three vaccinia-naïve subjects were randomized to receive two vaccine doses, either lyophilized MVA subcutaneously (SC), liquid MVA SC or liquid MVA intradermally (ID) 28 days apart. For a subset of subjects, antibody-dependent cellular cytotoxicity (ADCC), IFN- γ release enzyme-linked immunospot (ELISPOT), and protein microarray antibody-binding assays were conducted. Protein microarray responses were assessed for correlations with PRNT, enzyme-linked immunosorbent assay (ELISA), ADCC and ELISPOT results. Results MVA elicited significant microarray antibody responses to 15 of 224 antigens, mostly virion membrane proteins, at day 28 or 42, particularly, WR113/D8L and WR101H3L. In the Liquid-SC group, responses to nine antigens, including WR113/D8L and WR101/H3L, correlated with PRNT results. Three correlated for the Liquid-ID group. No significant correlations were observed with ELISPOT responses. In the Liquid-ID group, WR052/F13L, a membrane glycoprotein, correlated with ADCC responses. Conclusions MVA elicited antibodies to fifteen vaccinia strain antigens representing virion membrane. Antibody responses to two proteins strongly increased and significantly correlated with increases in PRNT. Responses to these proteins are potential correlates of protection and may serve as immunogens for future vaccine development.

scholarly journals Evaluation of a Pseudotyped Virus Neutralisation Test for the Measurement of Equine Influenza Virus-Neutralising Antibody Responses Induced by Vaccination and Infection

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 466
Author(s):  
Rebecca Kinsley ◽  
Stéphane Pronost ◽  
Manuelle De Bock ◽  
Nigel Temperton ◽  
Janet M. Daly ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Haemagglutination Inhibition ◽  
Antibody Responses ◽  
Pseudotyped Virus ◽  
Equine Influenza Virus ◽  
Serum Samples ◽  
Neutralising Antibodies ◽  
Protective Antibody ◽  
Equine Influenza ◽  
Antibody Assays

Equine influenza is a major respiratory disease of horses that is largely controlled by vaccination in some equine populations. Virus-neutralising antibodies, the mainstay of the protective immune response, are problematic in assaying for equine influenza virus, as most strains do not replicate efficiently in cell culture. Surrogate measures of protective antibody responses include the haemagglutination inhibition (HI) test and single radial haemolysis (SRH) assay. For this study, a pseudotyped virus, bearing an envelope containing the haemagglutinin (HA) from the Florida clade 2 equine influenza virus strain A/equine/Richmond/1/07 (H3N8), was generated to measure HA-specific neutralising antibodies in serum samples (n = 134) from vaccinated or experimentally-infected ponies using a pseudotyped virus neutralization test (PVNT). Overall, the results of PVNT were in good agreement with results from the SRH assay (100% sensitivity, 68.53% specificity) and HI test (99.2% sensitivity, 49.03% specificity). The PVNT was apparently more sensitive than either the SRH assay or the HI test, which could be advantageous for studying the antibody kinetics, particularly when antibody levels are low. Nevertheless, further studies are required to determine whether a protective antibody level can be defined for the SRH assay and to ascertain the inter-laboratory reproducibility. In conclusion, the PVNT efficiently measures neutralising antibodies after immunization and/or experimental infection in the natural host, and may complement existing antibody assays.

scholarly journals Neutralization Fingerprinting Technology for Characterizing Polyclonal Antibody Responses to Dengue Vaccines

2021 ◽  
Author(s):  
Nagarajan Raju ◽  
Xiaoyan Zhan ◽  
Subash Das ◽  
Lovkesh Karwal ◽  
Hansi J. Dean ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Vaccine Development ◽  
Dengue Infection ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Modeling Framework ◽  
Correlates Of Protection ◽  
Immune Correlates ◽  
Tetravalent Vaccine ◽  
Antibody Specificities

AbstractDengue is a major public health threat. There are four serotypes of dengue virus (DENV), therefore efforts are focused on development of safe and effective tetravalent DENV vaccines. While neutralizing antibodies contribute to protective immunity, there are still important gaps in understanding of immune responses elicited by dengue infection and vaccination, including defining immune correlates of protection. To that end, here we present a computational modeling framework for evaluating the specificities of neutralizing antibodies elicited to tetravalent DENV vaccines, based on the concept of antibody-virus neutralization fingerprints. We developed and applied this framework to samples from clinical studies of TAK-003, a tetravalent vaccine candidate currently in phase 3 trials, to characterize the effect of prior dengue infection (baseline) on the specificities of vaccine-elicited antibody responses. Our results suggested a similarity of neutralizing antibody specificities in baseline-seronegative individuals. In contrast, amplification of pre-existing neutralizing antibody specificities was predicted for baseline-seropositive individuals, thus quantifying the role of immunologic imprinting in driving antibody responses to DENV vaccines. The analysis framework proposed here can apply to studies of sequential dengue infections and other tetravalent DENV vaccines and can contribute to understanding dengue immune correlates of protection to help guide further vaccine development and optimization.

scholarly journals SARS-CoV-2 antibody magnitude and detectability are driven by disease severity, timing, and assay

2021 ◽  
Author(s):  
Michael J. Peluso ◽  
Saki Takahashi ◽  
Jill Hakim ◽  
J. Daniel Kelly ◽  
Leonel Torres ◽  
...  
Keyword(s):  
Disease Severity ◽  
Antibody Responses ◽  
Binding Assays ◽  
Oxygen Requirement ◽  
Test Characteristics ◽  
Clinical Presentations ◽  
Antibody Assays ◽  
Previous Infection ◽  
Antibody Titers ◽  
Infection Timing

ABSTRACTSerosurveillance studies are critical for estimating SARS-CoV-2 transmission and immunity, but interpretation of results is currently limited by poorly defined variability in the performance of antibody assays to detect seroreactivity over time in individuals with different clinical presentations. We measured longitudinal antibody responses to SARS-CoV-2 in plasma samples from a diverse cohort of 128 individuals over 160 days using 14 binding and neutralization assays. For all assays, we found a consistent and strong effect of disease severity on antibody magnitude, with fever, cough, hospitalization, and oxygen requirement explaining much of this variation. We found that binding assays measuring responses to spike protein had consistently higher correlation with neutralization than those measuring responses to nucleocapsid, regardless of assay format and sample timing. However, assays varied substantially with respect to sensitivity during early convalescence and in time to seroreversion. Variations in sensitivity and durability were particularly dramatic for individuals with mild infection, who had consistently lower antibody titers and represent the majority of the infected population, with sensitivities often differing substantially from reported test characteristics (e.g., amongst commercial assays, sensitivity at 6 months ranged from 33% for ARCHITECT IgG to 98% for VITROS Total Ig). Thus, the ability to detect previous infection by SARS-CoV-2 is highly dependent on the severity of the initial infection, timing relative to infection, and the assay used. These findings have important implications for the design and interpretation of SARS-CoV-2 serosurveillance studies.

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