Modelling SARS-CoV-2 infection in a human alveolus microphysiological system
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The coronavirus 2019 pandemic has highlighted the importance of physiologically relevant in vitro models to assist preclinical research. Here, we describe the adaptation of a human alveolus microphysiological system (MPS) model consisting of primary human alveolar epithelial and lung microvascular endothelial cells to study infection with SARS-CoV-2 at Biosafety Level 3 facility. This infection model recapitulates breathing-like stretch and culture of epithelial cells at the air–liquid interface and resulted in clinically relevant cytopathic effects including cell rounding of alveolar type 2 cells and disruption of the tight junction protein occludin. Viral replication was confirmed by immunocytochemical nucleocapsid staining in the epithelium and increased shedding of SARS-CoV-2 virus within 2 days post-infection, associated with changes in innate host immune responses. Together, these data demonstrate that, under the experimental conditions used in this work, this human alveolus MPS chip can successfully model SARS-CoV-2 infection of human alveolar lung cells.
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Thank you for the submission of your revised manuscript and addressing the concerns raised by the reviewers.
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The reviewers have raised some concerns, pleaase make sure you address them carefully. In particular those that relate to clarity, data visualisation as well as inclusion of statistical analysis data (reviewer 2 lines 195-197).
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Comments to Author
This manuscript describes the adaptation of a human alveolus microphysiological system (MPS) model consisting of primary human alveolar epithelial and lung microvascular endothelial cells to study infection with SARS-CoV-2. In general, the data is well presented and reflects the innovation and advantages of using this model. However, while reviewing this work one major critique was identified. Furthermore, several minor critiques should also be addressed to provide clarity to the text. These critiques are outlined below. Mayor critique: - Given that the main contribution of this manuscript is the description of a microphysiological system to study infection with SARS-CoV-2, it is essential to include a figure where this model is extensively described. This will allow the reader get familiarized with …
Comments to Author
This manuscript describes the adaptation of a human alveolus microphysiological system (MPS) model consisting of primary human alveolar epithelial and lung microvascular endothelial cells to study infection with SARS-CoV-2. In general, the data is well presented and reflects the innovation and advantages of using this model. However, while reviewing this work one major critique was identified. Furthermore, several minor critiques should also be addressed to provide clarity to the text. These critiques are outlined below. Mayor critique: - Given that the main contribution of this manuscript is the description of a microphysiological system to study infection with SARS-CoV-2, it is essential to include a figure where this model is extensively described. This will allow the reader get familiarized with the model and to easily understand the data shown in the results section. Minor critiques: - Lines 156-159: It is not clear how the authors manipulated stretch conditions in this model. Since this characteristic constitutes a major innovation in this model, it is recommended to use a figure (listed above as a major critique) to explain how stress is introduced and manipulated in this model. - Figure S2: Use arrows/arrowheads/asterisks to indicate rounding of the infected cells, cell aggregation, clumping, and increased cytoplasmic vacuolization in epithelial cells. - Lines 168-169: It is recommended to change this sentence for: by staining alveolus-on-chips with antibodies specific for HT2-280, marker of AT2, and tight junction protein OCLN". - Line 176: Add references. - Figure 2: Indicate the size of the scale bar. Play with the contrast of the image or add a higher magnification image to appreciate better the increased detection of SARS-CoV-2 nucleocapsid through time. - Lines 195-197: Perform the appropriate statistical analysis and include the results in this section and in Figure 2. - Lines 228-230: Add a reference to Figure 3b. - Line 280: It is recommended to change this sentence for: "a chemoattractant for monocytes, TRAIL which induces apoptosis, and to". - Lines 283-286: It is recommended to change this sentence for: In contrast, IL-8 and to some extent IL-6, which are associated with disease severity of COVID-19 [33] and have shown to be increased in previous models [3, 4], showed a reduction as a response to SARS-CoV-2 infection in our model. This effect may be due to the absence of immune cells in our model.
Please rate the manuscript for methodological rigour
Good
Please rate the quality of the presentation and structure of the manuscript
Good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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Comments to Author
This is a well written paper which describes validation of an MPS model for lung tissue infected with SARS-CoV-2. While generally reasonable in conclusions and goals there remains some issues to address. The authors point out the discrepancies between the cytokine responses observed in this study and comparable studies (lines 283-286) and suggest that this is connected to lack of immune cells in the model described here. While this is plausible this is also a critical observation for this study and a reasonable discussion should expand on this point. This is particularly important given the study seeks to demonstrate that the model recapitulates key clinical features, of which comparable cytokine responses must be considered a critical component. There are a number of issues with presentation of …
Comments to Author
This is a well written paper which describes validation of an MPS model for lung tissue infected with SARS-CoV-2. While generally reasonable in conclusions and goals there remains some issues to address. The authors point out the discrepancies between the cytokine responses observed in this study and comparable studies (lines 283-286) and suggest that this is connected to lack of immune cells in the model described here. While this is plausible this is also a critical observation for this study and a reasonable discussion should expand on this point. This is particularly important given the study seeks to demonstrate that the model recapitulates key clinical features, of which comparable cytokine responses must be considered a critical component. There are a number of issues with presentation of figures and formatting that should be addressed prior to publication. Figure 1: seems to be a low quality image, a higher resolution image will be needed for publication. The current image is difficult to interpret. Figure 2: part A should be edited to be formatted uniformly. Figure 3: Panels A and B should be made uniform i.e. the time points on the X axis are on both rows for panel B but only displayed on the bottom row of panel A.
Please rate the manuscript for methodological rigour
Satisfactory
Please rate the quality of the presentation and structure of the manuscript
Satisfactory
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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