PONE-D-20-06818

Cerebrospinal fluid oligoclonal bands in Neuroborreliosis are specific for Borrelia burgdorferi

PLOS ONE

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'Borrelia antigens were provided free of charge by Euroimmun.

URL: https://www.euroimmun.com/startseite.html

The funders had no role in study design, data collection and analysis, decision to

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Additional Editor Comments (if provided):

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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

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

Reviewer #1: Yes

Reviewer #2: 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

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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

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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: The authors established a test to detect, whether CSF OCBs in patients with neuroborreliosis (NB) are specific for borrelia burgdorferi senso lato. By applying an immunoblotting test, the authors suggest that OCBs in LN are indeed specific against borrelia burgdorferi. The major limitation of this study is the limited number of patients with LN (n=10) and controls (n=11), however, the authors state in the discussion, that this is an exploratory study.

Limitations

What antigens for borrelia burgdorferi are available and what borrelia antigen has been used for pre-coated NCM (page 4, line 85)? The author could shortly refer to antigens (also in comparison to ELISA tests) and their specificity – also in regard to cross reactivities (maybe see Chandra A et al., 2011 Epitope mapping…).

The authors could shortly summarize common borrelia specific tests and their sensitivity / specificity (maybe as a table). For example CSF CXCL13 has been suggested as a clinical test to detect NB, although CXCL13 is not specific for NB (Rupprecht et al., 2018, Diagnostic value of CSF CXCL13…; Lepennetier et al, 2019, Cytokine and immune cell…).

What is the advantage of the new test?

Could the test be introduced in a clinical setting to improve diagnostics?

Why do the authors apply a cut-off of > 2 bands for the pre-coated NCM test? Although OCBs are usually considered positive when >2 bands are specifically detected in CSF, why should this approach by applied for the immunoblotting test, especially in the context of the positive control? Please reconsider this issue.

It would be interesting, if the control patient (page 9, line 181) with positive OCBs in the immunoblotting test has a positive antibody titer – at least in serum.

Table 1: CSF standard finding could be more detailed (e.g. Ig index, intrathecal Ig production, glucose. Lactate …)

Picture quality of Figure 1A right is difficult which may be due to submission format.

It would have been interesting to correlate CXCL13 values (as a measure of B cell activity) with OCBs and test results.

Reviewer #2: In this retrospective, cross-sectional case-control study CSF-serum sample pairs of 21 patients (10 with neuroborreliosis and 11 OCB positive controls) were included. The authors aimed to determine whether CSF OCBs in patients with neuroborreliosis are specific for borrelia burgdorferi. Isoelectric focusing followed by immunoblotting using (A) an uncoated as well as (B) a borrelia antigen pre-coated nitrocellulose membrane was performed. The results indicate borrelia-specific bands in 9 of 10 neuroborreliosis patients. The manuscript is well written and concise.

It might profit in view of the reviewer from additionally addressing a few minor points:

(1) Methods section: please indicate that CSF-serum sample pairs were analyzed.

(2) Figure 1 could be extended by another example C (OCB type 2, e.g., MS CSF-serum

pair).

(3) Please discuss the possible applications of this test method. Are there any

advantages with regard to the current standard (sample volume, costs, ..)?

(4) In table 2: Are the data normally distributed? If not, please indicate the median

and range or IQR. Please indicate the OCB positivity rate in each group. Visual analysis

NB uncoated 100 vs pre-coated 90%, Controls 100 vs 0%; photometric analysis NB 100 vs 90%

(?), Controls 90,9% vs

0% (?).

(5) Not sure whether the sample size is sufficient to make statements on diagnostic

accuracy, although the results are conclusive. This could be determined in a larger study?

More from academic interest than practical relevance: Is there a correlation between

anti-borrelia specific antibody indices and the number of borrelia specific OCBs?

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Reviewer #1: No

Reviewer #2: No

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Cerebrospinal fluid oligoclonal bands in Neuroborreliosis are specific for Borrelia burgdorferi

PONE-D-20-06818R1

Dear Dr. Deisenhammer,

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,

Edgar Meinl, M.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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: All comments have been addressed

Reviewer #2: 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: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: 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

**********

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

**********

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: All comments have been addressed sufficiently, and there are no further questions. The manuscript provides an interesting view on OCBs in Lyme neuroborreliosis.

Reviewer #2: I thank the authors for responding to my prior suggestions.

All comments have been addressed.

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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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Reviewer #1: No

Reviewer #2: No