Cord Blood Levels of EPA, a Marker of Fish Intake, Correlate with Infants’ T- and B-Lymphocyte Phenotypes and Risk for Allergic Disease

Round 1

Reviewer 1 Report

LCPUFA > long chain polyunsaturated fatty acids (LCPUFA) in abstract

An important aspect of the paper is the PCA showing the same clusters for allergy at 3 years and allergy at 8 years. The paper says they were not necessarily the same subjects. Is there something that differentiates the age groups. Perhaps food allergy vs aeroallergy.

No breakdown of aeroallergy and food allergy or information on the proportion of aeroallergy and food allergy -small sample but at least give some idea.

“Other manifestations of allergy” in Table 1 is a bit loose. Are these clinical manifestations?

Need some explanation and methodology for OPLS models and the difference with the 3 year association of allergy with BAFF between the OPLS and PCA  analyses

Author Response

Please see PDF file

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript is well written and the conclusions drawn are supported by the data. The results are very interesting and potentially important for human health and disease intervention. My specific comments are listed below.
- Line 24: abstract do not explain why farming families are compared to control one. Please, consider adding more information to abstract
- Line 29: do authors expect/consider any other components besides LCPUFA responsible for this observation?
- Line 50: please add ‘AA’ in bracket -> (AA, 20:4 n-6) to maintain consistency in abbreviations (see line 44)
- Line 57: ‘naïve B cells’, please correct the spelling
- Line 62: please add an explanation of Tregs -> Tregs (Regulatory T cells)
- Line 78: ‘Sixty-three children (97%)’ please specified how many from each group
- Line 79: same as 78. is the median of age important here? Why it wasn’t important during a 3-year follow-up?
- Line 86: please specified types of foods,
- Line 87: Please specified types of inhalant allergens
- Table 1: double category naming, please correct 18m as 1,5year for standardization
- Line 108: from how many patients' dietary questionnaires were obtained? How many from each group?
- Line 119: in the subject section authors wrote about 65 families, here authors mention 54 samples, please explain the difference
- Line 136,137: an unnecessary explanation of abbreviations
- Line 143: how many samples? Same number as in section 2.4? How many from each group?
- Line 145: supplementary table 1 was not provided to the reviewer
- Line 155-159: please add a brief description of the method, how many samples were analyzed, how many from each group
- Line 180-181: ‘In brief, 64, 63 and 48 181 infants participated in the follow-ups at 1.5, 3 and 8 years, respectively, and 15, 11 and 10 infants were 182 diagnosed as allergic’ how many from each group (farming vs control)
- Line 182: please specify how many had ARC, asthma, eczema, and food allergy (within groups)
- Line 194: what were the levels of cord blood LCPUFA? What were the maternal fish intakes? please consider presenting the results from the dietary questionnaire and GC-MS
- Line 204: DPA - please add abbreviation meaning
- Line 207: please explain the sentence: ‘It is noteworthy that while the proportion of DPA was related to that of DHA, and the total n-3 LCPUFAs, i.e., they clustered together, this was not true for the 209 proportion of EPA’ please consider rephrasing the sentence
- Figure 2: figure 2 will be easier to read and compare when both panels will be on the same page
- Line 252: ‘fig. 3’ fig3a or fig3b or both, please specified as inline 255
- Line 262: ‘Thus, five children were allergic at both time-points, while four were only allergic at 3 years’ so 9 children at 3yr, there were 11 children in section 3.1, please explain
- Line 273: ‘Thus, 55 infants were included in the 274 OPLS model for allergy at 3 years of age’ Why 55? According to section 3.1, there were 63 children and 6 had a transient allergy. Please explain
- Line 284: “We hypothesized that components of fish in the maternal diet’ – what components? Do you consider any other component besides PUFA? Please consider rephrasing this statement
- Line 420: ‘Another strength and novel aspect of this study is the application of biomarkers of fish intake’ explain the novelty if EPA is a recognized biomarker

Author Response

Please see PDF file

Author Response File: Author Response.pdf

Reviewer 3 Report

This is an interesting and important paper regarding the influence of fish consumption during pregnancy on the sensitization outcome in the children, followed until 8 years of age. The paper is excellently written, clearly structured and contains all relevant references. Also, limitations of the study, e.g. small sample size have clearly been stated.

I have only few important comments:

1. a scheme of the study groups / population / incl. results would help, e.g. starting point is XXX farming children vs. YYYY non-farming children; among them XXX allergic vs. non-allergic; n=allergic at 1.5 years/3 years/8 years; e.g. for clarification of lines 262 ("five children were allergic at both time-points" vs. Fig.3-legend "transient allergy" vs. "55 infants were included")
2. very important: was the farming/non-farming situation acknowledged in the stat. analysis? e.g. in Fig.2 this should be included as an additional factor
3. please give a short description of the differences in study groups instead of having this important info in reference only (line 190).

Minor: 

• I could not retrieve Suppl Table 1
• what is new here compared to previous studies needs to be clearly stated in Intro (lines 46 vs. lines 64)
• line 77: "weeks" small letters
• Table 1: instead of "18m" use "1.5 yr"; Williams criteria could use reference; at the end : "FEV1 ≥ 12%" seems to be typo included
• "airway rhinoconjunctivitis" is an unknown term to me
• line 101: "food allergy symptoms" (or list some of them)
• line 131: "of 4°C" (insert space)
• line 191: was smoking (passive) included in comparison?
• Fig. 4/in heat-map: used Greek letters for a4b7 labelling
• line 370: "possible mechanisms" have not been shown, just influence factors (like fish consumption and /or certain fatty acids; but not mechanisms)

Author Response

Please see PDF file

Author Response File: Author Response.pdf