Development of a Novel Chimeric ND-GP cVLPs Vaccine for the Prevention of Goose-Derived Newcastle Disease and Gosling Plague

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript details the creation of a significant bivalent chimera (ND-GP cVLPs) that presents the NDV HN protein and the GPV VP3 protein. This innovative approach has led to the development of a potent vaccine for the Newcastle disease virus. The research findings demonstrate that when immunized with 30-70 μg of ND-GP cVLPs, geese produced highly effective hemagglutination inhibitory and neutralizing antibodies.

The manuscript provides crucial information, and the experimental steps were meticulously conducted and described. The introduction, while comprehensive, could be condensed for brevity. The MM and results are presented, and the figures are necessary and sufficient. The manuscript deserves publication, and we strongly recommend an English language upgrade to significantly enhance its presentation and quality. Abstract, but this should be done throughout the text,

Traditional- change to “conventional.”

In vivo-add “coma” In vivo,

Furthermore- add “coma.”

Due to- change to “from”

The use of-change to “using”

The development- change to “developing.”

Comments on the Quality of English Language

The English is understandable but could be improved to increase the manuscript's impact.

 

Author Response

Reviewer #1:

Comment 1: The manuscript provides crucial information, and the experimental steps were meticulously conducted and described. The introduction, while comprehensive, could be condensed for brevity. The MM and results are presented, and the figures are necessary and sufficient. The manuscript deserves publication, and we strongly recommend an English language upgrade to significantly enhance its presentation and quality. Abstract, but this should be done throughout the text。Traditional- change to “conventional.”, In vivo-add “coma” In vivo, Furthermore- add “coma.”, Due to- change to “from”, The use of-change to “using”,The development- change to “developing.”

Comments on the Quality of English Language

The English is understandable but could be improved to increase the manuscript's impact.

Response: Thank you very much for your suggestions. We have reviewed the manuscript and corrected the language errors. We have also condensed the introduction descriptions. The revised portions have been marked in red in the revised manuscript.

Reviewer 2 Report

Comments and Suggestions for Authors

In the Newcastle Disease (ND) and Gosling Plague (GP) outcome studies, the authors of the current publication employed GP VP3 protein and ND HN protein to construct ecofriendly and efficient chimeric ND-GP bivalent virus-like particles. Furthermore, they investigated the effects of immunization and challenge of goslings, as well as the levels of NDV HI antibodies and GPV-neutralizing antibodies. The methodology is soundly planned and executed. The methodology is adequately described and the results are clearly presented, thus addressing any concerns I may have. The work has significant epidemiological implications.  
Specific comments:
2.2, 2.3 – Please provide a detailed account of the methodology employed to ascertain the specificity of the GPV VP3 mouse polyclonal antibody.
2.7 - Please indicate whether the data were tested for normal distribution and, if so, which test was used.
2.8 - Please indicate the number of ethical committee approvals obtained to perform the experiment.
Figure 5 – No methodological description of how the histopathological preparations were made is provided in the text of the paper. It is unclear from which part of the intestine the samples came and from which part of the brain.

Author Response

Reviewer #3:

Comment 1: 2.2, 2.3 – Please provide a detailed account of the methodology employed to ascertain the specificity of the GPV VP3 mouse polyclonal antibody.

Response: The VP3 antibody was prepared by targeting the dominant B-cell epitope region of the VP3 protein. The immunogen information is as follows: NEVDSSRNAQFKKAVKGAYGTMGRNWLPGPKFLDQRVRAYTGGTDNYANWNIWSNGNKVNLKDRQYLLQPGPVSATYTEGEASSLPAQNILGIAKDPYRSGSTTAGISDIMVTEEQEVAPTNGVGWKPYGRTVTNEQNTTTAPTSSDLDVLGALPGMVWQNRDIYLQGPIGAKIPK. Firstly, we performed a BLAST search in the NCBI database, comparing the immunogen with the proteomes of insect cells, chicken, and goose. The results did not reveal any similar proteins, thus confirming the specificity of the antibody for the immunogen (Figure 1A-C). Furthermore, we have conducted a high-purity purification of the immune source protein (Figure 1D). Additionally, the ELISA titer of the collected antibodies is guaranteed to be higher than 1:128,000. The Western blot results also confirm that this antibody does not show non-specific binding bands (as shown in Figures 1C and 2C of the manuscript).

Figure 1. Specificity of GPV VP3 antibody. Alignment results of the immunogen with reference proteins from insects (A), chickens (B), and geese (C). (D) SDS-PAGE identification results of the purified immunogen.

 

Comment 2: 2.7 - Please indicate whether the data were tested for normal distribution and, if so, which test was used.

Response: Since the sample size is not large, we used the Shapiro-Wilk test to assess the normal distribution of the data, which has been supplemented and marked in red in the revised manuscript.

 

Comment 3: 2.8 - Please indicate the number of ethical committee approvals obtained to perform the experiment.

Response: We apologize for any incomplete information in our description. The animal ethics were approved by the Animal Welfare and Research Ethics Committee of Jilin University, with Approval Code: KT202302225. The revised manuscript has been updated and marked in red.

 

Comment 4: Figure 5 – No methodological description of how the histopathological preparations were made is provided in the text of the paper. It is unclear from which part of the intestine the samples came and from which part of the brain.

Response: We have added the relevant methods for pathological sectioning in "2.6 Detection of the protective effect against challenge" and provided detailed descriptions of the brain and intestine extraction sites. The small intestine samples were obtained from the duodenum, and the brain samples were taken randomly from the cerebrum.

Author Response File: Author Response.pdf