Review for 10.1186/s12876-023-02798-0 (2023)

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".
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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