Reviewer’s report
Title: 
Peripheral CD4+CD25hiCD127low regulatory T cells are increased in
patients with gastrointestinal cancer
Version: 
0 
Date: 
14 Jul 2022
Reviewer's report:
Qiu et al. have submitted a study of levels of CD4+CD25hiCD127low Tregs in
patients with gastrointestinal cancer. They enrolled 45 gastric cancer patients, 50
colorectal cancer patients and 50 healthy controls over 4 years. They found the
levels of CD4+CD25hiCD127low Tregs and CD4+CD25hi cells increased
significantly in patients with gastrointestinal cancer, as well as an elevated levels
of IL-10 and TGF-
β
1 both in their serum and seperated CD4+CD25hiCD127low
Tregs. It is an important issue to illustrate the immunological features of
gastrointestinal patients. However, some revision and clarifications are needed. 
Major comments
1.What was the demographic information about Healthy control, any
comorbidities? The authors have stated that "the clinical characteristics of all the
participants were detailed in Table 1 (Line127-128)", but there is a lack of the
information about Health control group in Table 1. 
2.Whether the tests of Flow cytometry and ELISA were conducted using
individual samples rather than pooled samples? If so, please clarify in the
Methods. Meanwhile, the column chart of Figure 2 and 3 were encouraged to be
changed to scatterplots which can show the value of each samples clearly. Figure
2A, the Y-axis should be labled as "% of CD4+ cells/lymphocytes" to make it
clear.
3.How does the differentiation (Poor, Moderate, High) and stages (early, late) of
GC and CRC "confirmed", it should be clarified in Methods or in the appropriate
section.
4.The serum levels of TGF-
β
 detected by ELISA (even in healthy donors) was
reported in the magnitude of tens or hundreds of ng/ml, while the authors only
detected no more than 30pg/ml TGF-
β
 in serum from each group, highly
recommended authors to double-check the original data to make sure the results
showed in Table 3 and Table 4 correctly.
5. It has been reported that high levels of CD4+ CD25hiCD127low cells were
related with early stage of several other cancers, does it also the case in
gastrointestinal cancer? Does high levels of CD4+CD25hiCD127low cells, IL-10
or TGF-
β
correlated with the differentiation or stages of GI, I believe these were
worth being examined and included in this study to make it more readable and
meaningful. 
Minor comments
1. Line106 "Comprehensively" and Line 108 "comprehensive" used twice in one
sentence, please use another replaceable word.
2. Line 110 "would providing" was incorrectly used. 
3. Line 111 "against patients with GI cancer" will be more appropriate to remove
the words "patients with". 
4. Line 448 "amongst in" might be "amongst".
Publons Reviewer Recognition
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find the details of processing in Publons’ privacy policy
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No
Declaration of competing interests
Please complete a declaration of competing
interests, considering the following questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the
editorial office before completing the review.
If you wish, you can include your
name in your published report. Please note you must decide whether to include
your name at the start of the process and confirm this decision whenever
prompted. If you change your decision later, we will not be able to publish your
name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
No
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
Yes
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
No
Reviewer’s report
Title: 
Peripheral CD4+CD25hiCD127low regulatory T cells are increased in
patients with gastrointestinal cancer
Version: 
0 
Date: 
14 Jul 2022
Reviewer's report:
Qiu et al. have submitted a study of levels of CD4+CD25hiCD127low Tregs in
patients with gastrointestinal cancer. They enrolled 45 gastric cancer patients, 50
colorectal cancer patients and 50 healthy controls over 4 years. They found the
levels of CD4+CD25hiCD127low Tregs and CD4+CD25hi cells increased
significantly in patients with gastrointestinal cancer, as well as an elevated levels
of IL-10 and TGF-
β
1 both in their serum and seperated CD4+CD25hiCD127low
Tregs. It is an important issue to illustrate the immunological features of
gastrointestinal patients. However, some revision and clarifications are needed. 
Major comments
1.What was the demographic information about Healthy control, any
comorbidities? The authors have stated that "the clinical characteristics of all the
participants were detailed in Table 1 (Line127-128)", but there is a lack of the
information about Health control group in Table 1. 
2.Whether the tests of Flow cytometry and ELISA were conducted using
individual samples rather than pooled samples? If so, please clarify in the
Methods. Meanwhile, the column chart of Figure 2 and 3 were encouraged to be
changed to scatterplots which can show the value of each samples clearly. Figure
2A, the Y-axis should be labled as "% of CD4+ cells/lymphocytes" to make it
clear.
3.How does the differentiation (Poor, Moderate, High) and stages (early, late) of
GC and CRC "confirmed", it should be clarified in Methods or in the appropriate
section.
4.The serum levels of TGF-
β
 detected by ELISA (even in healthy donors) was
reported in the magnitude of tens or hundreds of ng/ml, while the authors only
detected no more than 30pg/ml TGF-
β
 in serum from each group, highly
recommended authors to double-check the original data to make sure the results
showed in Table 3 and Table 4 correctly.
5. It has been reported that high levels of CD4+ CD25hiCD127low cells were
related with early stage of several other cancers, does it also the case in
gastrointestinal cancer? Does high levels of CD4+CD25hiCD127low cells, IL-10
or TGF-
β
correlated with the differentiation or stages of GI, I believe these were
worth being examined and included in this study to make it more readable and
meaningful. 
Minor comments
1. Line106 "Comprehensively" and Line 108 "comprehensive" used twice in one
sentence, please use another replaceable word.
2. Line 110 "would providing" was incorrectly used. 
3. Line 111 "against patients with GI cancer" will be more appropriate to remove
the words "patients with". 
4. Line 448 "amongst in" might be "amongst".
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons after the final decision on the manuscript has been made. If you have
already registered at Publons, they will notify you of the receipt of this review
and update your profile 
as per your settings and their policy. If you are not
registered with Publons, you will receive an email from them asking you to
register in order for them to be able to recognize your review on your new
profile page. Publons may use the Review Data to generate derivative metadata
for the benefit of Publons and you as a reviewer, carefully considering the
sensitivity of such information. For example, Publons may verify your record as
a reviewer by updating your profile published on its webservice if you have
registered for such service or help editors to identify candidate reviewers. Please
find the details of processing in Publons’ privacy policy
https://publons.com/about/terms
No
Declaration of competing interests
Please complete a declaration of competing
interests, considering the following questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the
editorial office before completing the review.
If you wish, you can include your
name in your published report. Please note you must decide whether to include
your name at the start of the process and confirm this decision whenever
prompted. If you change your decision later, we will not be able to publish your
name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
No
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
Yes
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
No
Please review this paper.
user
Reviewer’s report
Title: 
Peripheral CD4+CD25hiCD127low regulatory T cells are increased in
patients with gastrointestinal cancer
Version: 
0 
Date: 
14 Jul 2022
Reviewer's report:
Qiu et al. have submitted a study of levels of CD4+CD25hiCD127low Tregs in
patients with gastrointestinal cancer. They enrolled 45 gastric cancer patients, 50
colorectal cancer patients and 50 healthy controls over 4 years. They found the
levels of CD4+CD25hiCD127low Tregs and CD4+CD25hi cells increased
significantly in patients with gastrointestinal cancer, as well as an elevated levels
of IL-10 and TGF-
β
1 both in their serum and seperated CD4+CD25hiCD127low
Tregs. It is an important issue to illustrate the immunological features of
gastrointestinal patients. However, some revision and clarifications are needed. 
Major comments
1.What was the demographic information about Healthy control, any
comorbidities? The authors have stated that "the clinical characteristics of all the
participants were detailed in Table 1 (Line127-128)", but there is a lack of the
information about Health control group in Table 1. 
2.Whether the tests of Flow cytometry and ELISA were conducted using
individual samples rather than pooled samples? If so, please clarify in the
Methods. Meanwhile, the column chart of Figure 2 and 3 were encouraged to be
changed to scatterplots which can show the value of each samples clearly. Figure
2A, the Y-axis should be labled as "% of CD4+ cells/lymphocytes" to make it
clear.
3.How does the differentiation (Poor, Moderate, High) and stages (early, late) of
GC and CRC "confirmed", it should be clarified in Methods or in the appropriate
section.
4.The serum levels of TGF-
β
 detected by ELISA (even in healthy donors) was
reported in the magnitude of tens or hundreds of ng/ml, while the authors only
detected no more than 30pg/ml TGF-
β
 in serum from each group, highly
recommended authors to double-check the original data to make sure the results
showed in Table 3 and Table 4 correctly.
5. It has been reported that high levels of CD4+ CD25hiCD127low cells were
related with early stage of several other cancers, does it also the case in
gastrointestinal cancer? Does high levels of CD4+CD25hiCD127low cells, IL-10
or TGF-
β
correlated with the differentiation or stages of GI, I believe these were
worth being examined and included in this study to make it more readable and
meaningful. 
Minor comments
1. Line106 "Comprehensively" and Line 108 "comprehensive" used twice in one
sentence, please use another replaceable word.
2. Line 110 "would providing" was incorrectly used. 
3. Line 111 "against patients with GI cancer" will be more appropriate to remove
the words "patients with". 
4. Line 448 "amongst in" might be "amongst".
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons after the final decision on the manuscript has been made. If you have
already registered at Publons, they will notify you of the receipt of this review
and update your profile 
as per your settings and their policy. If you are not
registered with Publons, you will receive an email from them asking you to
register in order for them to be able to recognize your review on your new
profile page. Publons may use the Review Data to generate derivative metadata
for the benefit of Publons and you as a reviewer, carefully considering the
sensitivity of such information. For example, Publons may verify your record as
a reviewer by updating your profile published on its webservice if you have
registered for such service or help editors to identify candidate reviewers. Please
find the details of processing in Publons’ privacy policy
https://publons.com/about/terms
No
Declaration of competing interests
Please complete a declaration of competing
interests, considering the following questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the
editorial office before completing the review.
If you wish, you can include your
name in your published report. Please note you must decide whether to include
your name at the start of the process and confirm this decision whenever
prompted. If you change your decision later, we will not be able to publish your
name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
No
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
Yes
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
No
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