Review 3: "SARS-CoV-2 Antibodies Cross-React and Enhance Dengue Infection"

RR:C19 Evidence Scale rating by reviewer:

• Potentially informative. The main claims made are not strongly justified by the methods and data, but may yield some insight. The results and conclusions of the study may resemble those from the hypothetical ideal study, but there is substantial room for doubt. Decision-makers should consider this evidence only with a thorough understanding of its weaknesses, alongside other evidence and theory. Decision-makers should not consider this actionable, unless the weaknesses are clearly understood and there is other theory and evidence to further support it.

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Review:

This manuscript, authored by Jakhar and Sonar et al., presents an investigation into the potential cross-reactivity between SARS-CoV-2 antibodies and their capacity to exacerbate Dengue virus Type 2 infection. The primary emphasis of this study focuses around in-vitro experiments conducted in cell lines exhibiting monocyte morphology, characterized by a high expression of Fcreceptors. These experiments involved usage of human plasma samples from different waves of the SARS-CoV-2 pandemic or animal models sera post-immunization with SARS-CoV-2.

Furthermore, the researchers conducted an in-silico analysis of the antibody structures of the E protein dimer of DENV-2 and SARS-CoV-2 to identify epitope residues interaction. The significance of this research is of great importance due to the widespread impact of the SARSCoV-2 pandemic on large human populations residing in Dengue-endemic regions.

While the in-vitro data exhibits a notably good degree of robustness and applicability, a thorough examination of specific methodological details and addition of important controls still need to be warranted. The presentation of results was generally clear and transparent, though the computational analysis proved challenging to comprehend.

The paper's methodology for assessing the cross-recognition of the DENV-E protein by anti-Spike and Anti-RBD SARS-CoV-2 antibodies in-silico involved the utilization of antibodies with known structures acquired from the Protein Data Bank. The calculations are executed using less commonly employed software tools, which may raise significant questions for the typical readers interested in this field (mostly immunologist). To comprehend the underlying assumptions, hypotheses, and calculations pertaining to molecular dynamics, there is a need for substantial improvement in the clarity of the text, methods, and figure 4. As a reviewer, evaluating the validity of this approach posed a considerable challenge due to its distinctly unconventional nature and the absence of adequate explanations, making it difficult to arrive at a definitive judgment. Consequently, the paper leaves a substantial degree of uncertainty.

Taking in account these considerations, the cumulative findings of this study do not significantly enrich the current body of knowledge in the field. Consequently, the study does not warrant immediate publication and needs the inclusion of essential controls and would profit from the suggested issues. I respectfully advise the authors to attend to the specified points and figures. Furthermore, I identified several typographical errors in the text, and I recommend a language correction of the manuscript. Specifically, some Latin terms were not italicized, departing from conventional typographic practices.

Specific Points:

1. The co-incubation with hole plasma is for me problematic since there are several components included in the human plasma samples that can be between the donors highly divers and that might be having an effect on your infection efficacy. It would be a better practice, to repeat the experiments with at least some of the sera samples, to corroborate your results, but this time with a purified fraction of Antibodies from the hole plasma.

2. There is no any mechanistic inside on which is the binding partner. I would suggest to digest the M5B AB and see if the F(ab’)2, Fab or Fc are mediating the enhancement of infection.

The significance of this research is of great importance due to the widespread impact of the SARS-CoV-2 pandemic on large human populations residing in Dengue-endemic regions.