Attenuated Chimeric GI/GIII Vaccine Candidate against Japanese Encephalitis Virus
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors"Major corrections
The current discussion section seems like a repetition of the results section. The discussion could be well rewritten and discussing the importance of vaccine in outbreak control, citing and referring to more recent studies on vaccine development for other flavvivuses, and highlighting insights that could lead to perspectives.
Limitations could be discussed as near-future perspectives.
No p-value have been seen in the text while the authors included a statistical analysis section.
Lines 314- 316:The last section did not support the obtained data between the SA14-GI env group and the SA14-14-2 group. Statistical data could added to support this cconclusion.
#Minor corrections
Line 11: "In Asian regions" could be replaced by "In the Asian continent"
Line 47: "C proteins" could be replaced by " the Capsid (C) protein"
line 48: "Japanese encephalitis virus" could be abbreviated to "JEV" and in the following sections of the manuscript
Line 50: "genes" could be replaced by "protein" and anywhere else in the manuscript
Line 53: "currently, " could be replaced by "actually, "
Lines 55-57: the sentence could be rephrased to avoid repetition of "currently available". Authors could use "These vaccines"
Line 65: "moe" could be replaced by "more"
Lines 65-66: The sentence needs to be rephrase for more clarity
Lines 68-70: This sentence could be moved to the beginning of the section (line 59).
Line 112: "After 3 d, could be replaced by " 3 days"
Line 141: "ul" could be replaced by "uL" and anywhere else in the manuscript
Abbreviations could be in parenthesis at their first apparition in the text.
Lines 277-281: the sentence seems already in the introduction section. could the authors remove it?
Lines 313-314: the sentence could be rephrased for more clarity?
Comments on the Quality of English Language
Extensive editing of English language required
Author Response
Thank you for your comments. Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
Comments and Suggestions for AuthorsSome minor changes to the manuscript have been suggested
Comments for author File: 
Comments.pdf
minor changes are needed as outlined in the attached document
Author Response
Thank you for your comments. Please see the attachment.
Author Response File: Author Response.docx
Reviewer 3 Report
Comments and Suggestions for AuthorsThe Japanese encephalitis virus (JEV) is a non-segmented RNA virus, belonged to Flaviviridae family. It is transmitted through mosquitoes and has resulted in epidemic encephalitis. Till now, there are few JEV vaccines which mainly target the genotype III (GIII) strains but there is no currently available vaccine that targets currently dominant genotype I (GI) strains. Lee and colleagues made use of the reverse genetic system to generate a chimeric JEV as the vaccine candidate against JEV GI. This study is novel and appreciated, but several concerns must be further addressed and clarified. Some results are problematic.
Major concerns:
1. More precise description and wordings should be used.
a. Line44: “Three structural proteins (C, prM, and E) are the main infectious viral particles”, idea is not clear.
b. Line 48: “two envelope proteins (prM and E) derived from the host”, the description is incorrect.
c. Line 55: “live-recombinant vaccines”, idea is not clear.
d. Line 97: which genotype is SA-14 belonged to?
e. Line 106: “Syn_GI”, the description is not clear.
f. Line 107: “specific primers” have to be described.
g. Line 133: when was the blood serum harvested after two-time immunization in mice?
h. Line 158-160: cells were stained with crystal violet. What is the readout of the results? How was it quantified? Is a plate reader used?
i. Line 188: SA14-GI env plasmid was stated but it should be the plasmid encoding the full-length viral genome with GI E gene. The description is confusing. Same problem was found in Fig 1A. It is highly recommended to re-name the construct.
j. Line 210: Fig 3 indicates the viability and body weight loss of mice which were infected with different viruses. There was no direct evidence to demonstrate the neurovirulence of different viruses.
k. Line 238-239: “we inoculated mice immun- 238 ized twice with 1 × 105 PFU of each viral strain i.c. with the GI K05GS strain”, the idea is not clear.
l. Line 307: which 24 amino acid residues are involved?
2. Line 64: reference is required for the mutation rate.
3. Line 73-79: the advantage of vaccine should be put in the discussion but not the introduction.
4. In Fig 5, mice immunized with KO5GS had the strongest protection than those immunized with chimeric SA14-GI virus. Why was the live attenuated K05GS not used as the vaccine candidate? If this is the case, the rationale to develop the chimeric virus is questionable. Further discussion is required.
5. In Fig 3, mice inoculated i.c. with K05GS died in 7 days. However, in Fig 5, mice immunized with K05GS was used to examine the protective capacity. The result concerning the mice immunized with K05GS is problematic. Further discussion is required.
6. Line 309-311: what is the purpose of mentioning the chimeric JEV with 16 amino acid residues? Further discussion is required.
Comments on the Quality of English Language
Careful proof-reading is required, and there are some typo and grammatical errors.
Author Response
Thank you for your comments. Please see the attachment.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsNA
Comments on the Quality of English LanguageNA
Author Response
Thank you for reviewing this manuscript.
Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript is highly improved and my questions are addressed. I don't have any further comments.
Comments on the Quality of English LanguageMinor typo errors were found. Proof-reading is required before the publication.
Author Response
Thank you for reviewing this manuscript.
