Title: Correlation and Diagnostic Significance of CD4 T Cell Subsets and NLRP3
Inflammasome in Ulcerative Colitis: The Role of the NLRP3/T-bet/GATA3 Axis
Version: 4 Date: 09 Dec 2024
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
-While the manuscript specifies the number of UC (42) and control (30) subjects, there is no
mention of how these sample sizes were determined. For increased rigor, a power calculation
should be provided to justify the sample size, especially given the complexity of the immune
markers and the potential for inter-individual variation.
-The tissue processing steps, including the drying of colon specimens at 60°C, might need further
justification. High-temperature drying may affect protein preservation or conformation, which
could influence the results of IHC. It would be useful to mention whether this step was validated
or optimized for tissue preservation in previous studies or experiments.
-The methods section does not address potential confounding factors, such as medication use
(e.g., corticosteroids, immunosuppressants), comorbid conditions, or diet, that might influence
the expression of the immune markers studied. A brief mention of how such factors were
controlled for or accounted for would strengthen the study's design.
-The control group consists of patients with colon polyps, which is a reasonable choice for
comparing to UC patients. However, it would be beneficial to include a "healthy control" group
to establish a baseline for biomarker levels in individuals without any colonic pathology. This
would help to further isolate the effects of UC from those associated with any potential pre-
cancerous or benign polyp-related immune alterations.
-While the authors present a compelling correlation between immune markers and UC severity,
the mechanistic understanding of how the NLRP3 inflammasome regulates T-bet and GATA3,
and vice versa, could be expanded. The study mentions that NLRP3-P correlates with T-bet
and GATA3, but the precise molecular mechanisms behind this interaction remain unclear. A
more detailed exploration of how NLRP3 activation directly influences Th1/Th2 differentiation
or how it might affect the transcriptional activity of T-bet and GATA3 would add depth to the
discussion.
-Including a more detailed discussion on how IL-6 might influence the balance between Th1
and Th2 (e.g., by suppressing IFN-# and promoting GATA3) would enhance understanding of
these regulatory circuits in UC.
Title: Correlation and Diagnostic Significance of CD4 T Cell Subsets and NLRP3
Inflammasome in Ulcerative Colitis: The Role of the NLRP3/T-bet/GATA3 Axis
Version: 4 Date: 09 Dec 2024
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
-While the manuscript specifies the number of UC (42) and control (30) subjects, there is no
mention of how these sample sizes were determined. For increased rigor, a power calculation
should be provided to justify the sample size, especially given the complexity of the immune
markers and the potential for inter-individual variation.
-The tissue processing steps, including the drying of colon specimens at 60°C, might need further
justification. High-temperature drying may affect protein preservation or conformation, which
could influence the results of IHC. It would be useful to mention whether this step was validated
or optimized for tissue preservation in previous studies or experiments.
-The methods section does not address potential confounding factors, such as medication use
(e.g., corticosteroids, immunosuppressants), comorbid conditions, or diet, that might influence
the expression of the immune markers studied. A brief mention of how such factors were
controlled for or accounted for would strengthen the study's design.
-The control group consists of patients with colon polyps, which is a reasonable choice for
comparing to UC patients. However, it would be beneficial to include a "healthy control" group
to establish a baseline for biomarker levels in individuals without any colonic pathology. This
would help to further isolate the effects of UC from those associated with any potential pre-
cancerous or benign polyp-related immune alterations.
-While the authors present a compelling correlation between immune markers and UC severity,
the mechanistic understanding of how the NLRP3 inflammasome regulates T-bet and GATA3,
and vice versa, could be expanded. The study mentions that NLRP3-P correlates with T-bet
and GATA3, but the precise molecular mechanisms behind this interaction remain unclear. A
more detailed exploration of how NLRP3 activation directly influences Th1/Th2 differentiation
or how it might affect the transcriptional activity of T-bet and GATA3 would add depth to the
discussion.
-Including a more detailed discussion on how IL-6 might influence the balance between Th1
and Th2 (e.g., by suppressing IFN-# and promoting GATA3) would enhance understanding of
these regulatory circuits in UC.
Please review this paper.
user
Title: Correlation and Diagnostic Significance of CD4 T Cell Subsets and NLRP3
Inflammasome in Ulcerative Colitis: The Role of the NLRP3/T-bet/GATA3 Axis
Version: 4 Date: 09 Dec 2024
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
-While the manuscript specifies the number of UC (42) and control (30) subjects, there is no
mention of how these sample sizes were determined. For increased rigor, a power calculation
should be provided to justify the sample size, especially given the complexity of the immune
markers and the potential for inter-individual variation.
-The tissue processing steps, including the drying of colon specimens at 60°C, might need further
justification. High-temperature drying may affect protein preservation or conformation, which
could influence the results of IHC. It would be useful to mention whether this step was validated
or optimized for tissue preservation in previous studies or experiments.
-The methods section does not address potential confounding factors, such as medication use
(e.g., corticosteroids, immunosuppressants), comorbid conditions, or diet, that might influence
the expression of the immune markers studied. A brief mention of how such factors were
controlled for or accounted for would strengthen the study's design.
-The control group consists of patients with colon polyps, which is a reasonable choice for
comparing to UC patients. However, it would be beneficial to include a "healthy control" group
to establish a baseline for biomarker levels in individuals without any colonic pathology. This
would help to further isolate the effects of UC from those associated with any potential pre-
cancerous or benign polyp-related immune alterations.
-While the authors present a compelling correlation between immune markers and UC severity,
the mechanistic understanding of how the NLRP3 inflammasome regulates T-bet and GATA3,
and vice versa, could be expanded. The study mentions that NLRP3-P correlates with T-bet
and GATA3, but the precise molecular mechanisms behind this interaction remain unclear. A
more detailed exploration of how NLRP3 activation directly influences Th1/Th2 differentiation
or how it might affect the transcriptional activity of T-bet and GATA3 would add depth to the
discussion.
-Including a more detailed discussion on how IL-6 might influence the balance between Th1
and Th2 (e.g., by suppressing IFN-# and promoting GATA3) would enhance understanding of
these regulatory circuits in UC.
assistant