PONE-D-21-38887Maternal transfer of IgG sub-types and its association to Immunity against Plasmodium falciparum in a Ugandan birth cohort (Busia-Eastern Uganda)PLOS ONE

Dear Dr. Okek,

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Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: This study evaluated effects of Intermittent Prophylactic Treatment in Pregnancy and placental malaria status on malaria specific IgG levels in the fetus and the association between IgG levels and occurrence of malaria in the first year of the infants. This is an interesting and important research topic. I have some questions about the data analysis.

The authors mentioned that “Increasing antibody titer against P.falciparum may be more of a

marker of exposure to previous malaria infections”. What does “previous malaria” mean here? Mothers got malaria during pregnancy? Doesn’t placenta malaria capture that information?

For the survival analysis what if the placenta malaria status was used as the grouping variable instead of IgG levels?

Does “ANOVA in Box plot” means the data are plotted in box plot and comparisons were done using ANOVA? ANOVA compares means instead of medians. Because there are only two groups in the comparison, ANOVA is the same as t-test. Also many of the log-IgG level seem to have a skewed distribution, a non-parametric test may be needed. Due to the relatively large number of comparisons (4 IgG subtypes and 15 antigens) P-values need to be adjusted for multiple testing.

IgG levels may be affected by many factors, e.g., SP vs DP, placenta malaria status, clinical characteristics of the mothers. A multivariable regression model will be helpful to identify factors that may influence IgG levels.

In the result section about survival curves, hazard ratio (HR) were given at the end of the paragraph but which IgG was this? Does the star in Figure 3 means p<0.05 between the two groups? If so, stars were seen in most of the subplots but only a few HRs were mentioned in the text. In addition, it will be interesting to fit a Cox regression model to assess the association while adjusting other covariates in the model. The low concentration group had a lower risk comparing to the higher concentration group. Shouldn’t the HR be <1 rather than >1. Please double check the text/numbers.

Are data on the mother’s social economic status (SES) available? If so, it would be interesting to include the SES variables in the regression models that predicting IgG levels and the Cox model to see if they play a role.

Reviewer #2: The manuscript is very well written, and the results are very interesting. However, the authors could make some corrections.

Manuscript Title:

Authors can revise the title and mention the effect of treatment with SP and DP, instead of talking only about P. falciparum infection

Abstract

- Choose between antibody and anti-body

- Results: Specify the difference (high or low) in IgG4 between children from mothers on SP vs DP, same thing for EBA

- Conclusion: " ncreasing antibody titer against P.falciparum " in whom? Please specify

Introduction

- Antibody dependent cellular cytotoxicity (ADCC) - not antigen dependent cellular cytotoxicity

- Put space before the references or not. Please harmonize

- High expression of IgG4 has been associated with repeated exposure to specific…of IgG4: Please put reference

Materials and methods

- Participants and data collection: Give enough details

- Describe preparation of buffer A, then B

- MagPix multiplex bead array assay: If available, please provide the IgG positivity threshold and specify how this threshold was determined

- The sample collection times for carrying out the survival tests were not detailed.

Results

- Figure 1: Review the figure, with good resolution.

- Why does the representation of the graphs differ? Please harmonize

- Specify the number of participants per group (n = ….)

- Association of placental malaria and malaria specific IgG antibodies:

Is there not an effect of both treatment (IPTp) / placental infection with P.f on the level of IgG?

Did all women receive IPTp treatment?

The authors can compare the level of IgG in children from mothers on SP (P.f neg vs P.f pos), and in children from

mothers on PD (P.f neg vs P.f pos).

- Association of malaria specific IgG sub-types and incidence of malaria in the study population

Authors can change “probability of survival” to “probability of fewer occurrence of malaria”

Discussion :

The authors can provide explanations for the differences observed only with a few antigens. Do these antigens have any particularities? The results obtained must be well discussed.

Reviewer #3: Erick and colleagues presents a report investigating levels of IgG in pregnant women cord blood receiving IPTp (or not) and in their infants after birth. They further attempt to investigate if IgG levels in cord blood has a protective effect against malaria. Similar studies have been performed previously, but this study includes a broad panel of 15 different Pf IgG targets and does well to look at IgG subclasses.

Major comments

There are numerous grammatical errors throughout the text, figures, and tables. The authors need to have their manuscript reviewed by someone with English as their first language in order to provide a clearer presentation of their findings.

Minor comments

Abstract and throughout: make sure to specify that you are assaying only for IgG antibodies. For example, the phrase in the Abstract: "Cord concentrations of the erythrocyte binding antigens (EBA140, EBA175 and EBA181) were significantly different.." makes it sound like you are detection the actual Pf EBAs, and not the Ig against EBAs.

Throughout: the assay signal for IgG is never translated to a concentration (ug/mL for example), so the authors should not refer to "concentrations" of antibodies in their study. Could replace with a word like "levels"

Introduction:

- "Mandatory" for Ugandan women to be enrolled in IPTp?

- Can delete "and consumes most of this essential gas"

- Please modify "most importantly the role of IgG from plasma.." as Ig transfer is one of many protective factors and shouldn't be assigned as the most important

- Can delete "(total and sub-classes)" since this is redundant

- "These anti-malarial immunoglobulins have varied..."

- "Antibodies against merozoite surface proteins are the most expressed in cord blood". You can just say cord blood has high levels of anti-MSP antibodies.

- DP and SP need to be written out at first use.

Materials and Methods:

- Need to explain specifically how the blood samples were collected, stored short-term (what type of containers, coagulant, fractionated?), and temperature stored at long-term until the IgG assay. It appears plasma was the sample stored long-term.

- Provide company information for all lab chemicals and assay reagents

- Need to provide references for the antigens use in the panel, and more detail on coupling conditions.

- Need to provide software that was used for analysis, and how the Kaplan-Meir curves were created in the software

Results:

- Figure 1: why are all 4 panels not boxplots?

- Figure 1: Missing AMA1 for panel A

- Figure 1: is panel B for IgG2 or IgG3?

- Figure 3: need all x and y axes to be the same among panels and plots. Some x axes appear to be days, and some months. Y axis isn't "Probability of Survival" but "Probability of no malaria infection"

- Figure 3 legend: panel A is IgG1 correct?

Discussion:

- "so either of them can be used safely to prevent acquisition of malaria..." the safety of these drugs has nothing to do with IgG antibody levels in women taking them. Please revise.

- "We found no overall association between placental..."

- Please remove the sentence referencing hemozoin as it is incorrect and not relevant here.

- "cofounders such as IgGA"?

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6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PONE-D-21-38887R1Effects of anti-malarial prophylaxes on maternal transfer of Immunogloublin-G (IgG) and  association to Immunity against Plasmodium falciparum infections among Children in a Ugandan birth CohortPLOS ONE

Dear Dr. Okek,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised below during the review process.

Please submit your revised manuscript by Jun 04 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Ray Borrow, Ph.D., FRCPath

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Some of my previous questions/comments were not addressed in this revision which I will restate below. Due to the many factors that can influence malaria risk I still think a multivariate Cox regression is useful to understand the association.

A total of 16 Ig-G subtypes were used in the comparison and p-values need to be adjusted for multiple testing. I am not familiar with this particular research area, but would it be meaningful to combine all Ig-G subtypes into one variable in the analysis? This way multiple testing is not a concern and the main message will be easier to see. Right now there are too many comparisons which makes the result hard to interpret.

The conclusion from the survival model is that “antibody titer against P.falciparum may be more of a marker of exposure to previous malaria infections among infants from malaria endemic areas as opposed to their protective roles” which is interesting and that is why I recommended using placental malaria status as the grouping variable. If your conclusion is true I would expect to see higher malaria rates among those positive for placental malaria. The Cox model may also help to tease apart the effects of placental malaria and Ig-G levels.

Reviewer #2: The authors have taken into account the various comments and have done an excellent job in improving the manuscript.

Reviewer #3: The authors provide an improved version of the manuscript which is much easier to read and comprehend. A few minor issues need to be addressed:

- In Methods, the authors need to add company information for all of their chemicals, reagents, and other labware

- Concentrations of reagents such as secondary antibody and PE-conjugate used for the assays need to be included in Methods

- For Figures 1 and 2, include labels for the y axis on each panel. Perhaps on the x axis, can state "Placental Malaria", "No Placental Malaria", rather than "No", "Yes"

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Odilon Paterne NOUATIN

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PONE-D-21-38887R2Effects of anti-malarial prophylaxes on maternal transfer of Immunogloublin-G (IgG) and  association to Immunity against Plasmodium falciparum infections among Children in a Ugandan birth CohortPLOS ONE

Dear Dr. Okek,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised below during the review process.

Please submit your revised manuscript by Jul 29 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Ray Borrow, Ph.D., FRCPath

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

Reviewer #3: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Cox model: why were only subjects with >=3 episodes of fevers considered as a failure instead of occurrence of malaria as in Figure 4? How were subjects with 1 and 2 episodes handled in the model? Family socioeconomic status and placental malaria status need to be adjusted for in the cox model. There are too many IgG variables in the model. Are different IgG levels correlated? If not, maybe better to fit each IgG in the model in turn and adjust the p-values for multiple testing. Table 3 needs to show variable description instead of variables names used in the coding. What is the overall P value? Is it likelihood ratio test of all the variables in the model? The likelihood ratio test should test for all the IgG related variables only.

It is not clear what is Figure 3A showing. Do the three tick marks on the x-axis in each subplot denote different outcome? What is the y-axis? Birth weight cannot be close to 0. Also it is not clear how the 2-way ANOVA was set up? If birth weight of a child, time to first malaria incidence and number of malaria episodes are the outcomes and mother’s economic status is a factor what is the other factor?

Result section on Kaplan-Meir survival curve, why was it called “reciprocal hazard ratio”? Higher level was associated with higher risk. Shouldn’t the value presented just be the hazard ratio for higher levels of IgG?

Reviewer #2: (No Response)

Reviewer #3: The authors still need to arrange some elements for easier interpretation. Table 3 should have defined rows and columns (like Tables 1 and 2) and the first 5 rows ("Time variable...." to "Overall P...") need to be deleted. In Figure 3, the two plots need clear labeling on the x and y-axes. For A, I assume the y-axis is birthweight, but what are the three categories on the x-axis? For B, I assume the x-axis is time to first malaria episode?

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

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PONE-D-21-38887R3Effects of anti-malarial prophylaxes on maternal transfer of Immunogloublin-G (IgG) and  association to Immunity against Plasmodium falciparum infections among Children in a Ugandan birth CohortPLOS ONE

Dear Dr. Okek,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the last  points raised during the review process.

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Academic Editor

PLOS ONE

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Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

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Reviewer #1: (No Response)

Reviewer #2: Yes

Reviewer #3: Yes

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: “Placental malaria prevalence”- is this a binary (presence/absence) variable or a continuous variable?

Cox regression: “total IgG”- is the sum of IgG across IgG1, IgG2, IgG3 and IgG4? There are many IgG variables in the model and there may be collinearity. My suggestion is to include only IgG specific to one antigen at a time as a predictor in the cox regression while adjusting for other non-IgG covariates. The p-values of the IgG variables from all the different cox models should be adjusted for multiple testing.

Why wasn’t mother’s economic status included as a covariate in the model?

Cox regressions adjusting for potential confounders are more informative than Kaplan-Meir survival analysis. The section “Probability of survival to “probability of fewer occurrence of malaria”” seems to be superfluous.

The section “Effects of Economic status on birth outcomes and risk of malaria incidence”- the only association shown is with birth weight, which is not relevant to the main topic of this paper. If there is no association with malaria incidence or IgG levels, the entire section can go. Please go through the paper carefully and remove any language related to this analysis.

Reviewer #2: The authors have done an excellent job. I read with great interest the responses to the various comments from reviewers.

A minor error found in the “laboratory procedures”: 50μl of a secondary antibody specific for an IgG subtype (IgG1, IgG2, IgG2, IgG3 and IgG4)...IgG2 is repeated twice.

Manuscript can be accepted

Reviewer #3: Authors should attempt to further clean up their figures, and will need to submit publication-quality for the final version.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

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While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Effects of anti-malarial prophylaxes on maternal transfer of Immunogloublin-G (IgG) and  association to Immunity against Plasmodium falciparum infections among Children in a Ugandan birth Cohort

PONE-D-21-38887R4

Dear Dr. Okek,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Ray Borrow, Ph.D., FRCPath

Academic Editor

PLOS ONE

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