METHODS TO CHARACTERIZE ANTI-VIRAL IMMUNE RESPONSES REVEAL POTENT ADJUVANTS, NOVEL ANTIBODY GENES, AND SEROLOGICAL TECHNIQUES TO ADDRESS VIRAL OUTBREAKS
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2022
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Abstract
The COVID-19 pandemic has claimed millions of lives, and its causative agent, SARS-CoV-2, has infected hundreds of millions globally. The highly contagious nature of this respiratory virus has spurred massive global efforts to develop vaccines, therapeutics, and tests at record speeds. Recombinant subunit vaccine candidates take longer to produce than mRNA, which has delayed their availability. Additionally, identifying a suitable adjuvant is especially beneficial for this vaccine technology. Passive immunotherapeutics developed for the previous SARS outbreak (caused by SARS-CoV) were initially used while new monoclonal antibodies (mAb) were produced. These SARS-CoV-2 targeted mAb were sourced from convalescent plasma and mice while vaccines were being developed. Diagnostic tests to document viral infection were quickly developed and widely distributed early in the pandemic. Similarly, IgG and IgM testing for serosurveillance commenced almost immediately. However, resource-limited regions of the world were restricted in their access to these assays, resulting in minimal reporting of the dynamics of the pandemic in these regions.
To address these key aspects of responding to the COVID-19 pandemic, we conducted a study in BALB/c mice to identify a suitable adjuvant on readily available SARS-CoV-2 spike S1 model antigens in parallel with the development of a prefusion stabilized full-length spike vaccine candidate. We identified CoVaccine HTTM as producing robust Th1 and Th2 responses compared to Alhydrogel or S1 alone. We utilized samples from a Zika virus vaccine study in cynomolgus macaques for antibody discovery. We demonstrated that single-cell RNA sequencing (scRNA-seq) is an effective method for analyzing vaccine responses while collecting antibody V(D)J) genes for recombinant expression and development. We identified several neutralizing antibodies with this technique and revealed light chain V-genes not currently available in public databases. Lastly, we developed and deployed a multiplexed immunoassay (MIA) for serological analysis of exposure to SARS-CoV-2, NL63, Dengue, and Chikungunya viruses in Liberia, West Africa. This assay was also able to quantitate antibody concentrations to SARS-CoV-2. Furthermore, we adapted this assay for a multiplexed inhibition test (MINT) which could determine if serum antibodies could inhibit SARS-CoV-2 spike protein from binding its cognate receptor, human ACE-2, in a multiplexed fashion.
This work demonstrates methods to expedite responses to viral outbreaks through synergistic approaches in vaccine, immunotherapeutic, and assay development. Adjuvant testing in tandem with protein engineering allowed for a recombinant protein-based vaccine candidate to rapidly be tested in mice and macaques. Our methods for vaccine analysis stemming from MIA and scRNA-seq enabled single-cell transcriptome and paired heavy chain/ light chain antibody sequence resolution in a single experiment. These efforts may help expedite mAb development in future outbreaks and highlights the potential that the use of cynomolgus macaques may provide for rapid therapeutic antibody discovery and development. Lastly, our MIA and MINT assays developed to support vaccine development could perform effective serological studies in a resource-limited world region. This work, in total, has global health implications for addressing viral outbreaks.
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Immunology, Cellular biology, Antibodies, COVID-19, SARS-CoV-2, Serology, Vaccines
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87 pages
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