Title: Macrophage polarization toward M1 phenotype in T cell transfer colitis model
Version: 1 Date: 28 Oct 2023
Reviewer's report:
The manuscript from Ebihara et al. aimed to investigate the macrophage function in a T
cell transfer colitis murine model, and found that the macrophages polarized toward the M1
phenotype in the colonic LP. The polarized macrophages also activated the colitogenic T cells,
which may contribute to disease progression and could be a potential therapeutic target. This
manuscript is well-written, and elucidated the unappreciated role of macrophages in a T cell
transfer colitis model. This study has the potential to lead to several subsequent projects in
order to study more mechanisms and therapeutic measures. However, significant revisions are
suggested to improve the quality of the manuscript.
1. The presentation of flow cytometry data needs to be improved significantly. In figure 3,
instead of using exact numbers to calculate macrophages, inflammatory monocytes, neutrophils,
and T cells. The author should consider using their ratio to CD45+ cells (such as number of
macrophages/number of CD45+ cells), that would be a more objective way to present.
2. The gating strategies among figure 3 (F-I) and figure 3 (J-M) are different, the author should
standardized the gating, or explain why using different strategies. In order to quantify the
intracellular cytokines, it is advised to use Mean, or Median, Fluorescence Intensity (MFI).
3. Since CD11c is also a marker for DCs, and DCs are considered as innate immune cells,
the author may want to discuss how they differentiate DCs and M1 macrophages from CD11c
positive cells.
4. Regarding the histological score of colitis, it is better to list the individual score for epithelial
hyperplasia, goblet cell and crypt loss and inflammatory infiltration. This will help understand
which of the three criteria is the main contributer to the increased histological score. Arrows are
suggested to mark the different histological changes in panels of F, G and H from figure 1.
5. In spite the widely recognized role of T cells in IBD, more discussions on the role of
macrophages IBD onset and progression need be added.
6. Some discussion on the available therapeutic strategies targeting macrophage polarization
will be appreciated.
7. Figure 4 is pack with panel, it is advised to separate into a new figure.
Title: Macrophage polarization toward M1 phenotype in T cell transfer colitis model
Version: 1 Date: 28 Oct 2023
Reviewer's report:
The manuscript from Ebihara et al. aimed to investigate the macrophage function in a T
cell transfer colitis murine model, and found that the macrophages polarized toward the M1
phenotype in the colonic LP. The polarized macrophages also activated the colitogenic T cells,
which may contribute to disease progression and could be a potential therapeutic target. This
manuscript is well-written, and elucidated the unappreciated role of macrophages in a T cell
transfer colitis model. This study has the potential to lead to several subsequent projects in
order to study more mechanisms and therapeutic measures. However, significant revisions are
suggested to improve the quality of the manuscript.
1. The presentation of flow cytometry data needs to be improved significantly. In figure 3,
instead of using exact numbers to calculate macrophages, inflammatory monocytes, neutrophils,
and T cells. The author should consider using their ratio to CD45+ cells (such as number of
macrophages/number of CD45+ cells), that would be a more objective way to present.
2. The gating strategies among figure 3 (F-I) and figure 3 (J-M) are different, the author should
standardized the gating, or explain why using different strategies. In order to quantify the
intracellular cytokines, it is advised to use Mean, or Median, Fluorescence Intensity (MFI).
3. Since CD11c is also a marker for DCs, and DCs are considered as innate immune cells,
the author may want to discuss how they differentiate DCs and M1 macrophages from CD11c
positive cells.
4. Regarding the histological score of colitis, it is better to list the individual score for epithelial
hyperplasia, goblet cell and crypt loss and inflammatory infiltration. This will help understand
which of the three criteria is the main contributer to the increased histological score. Arrows are
suggested to mark the different histological changes in panels of F, G and H from figure 1.
5. In spite the widely recognized role of T cells in IBD, more discussions on the role of
macrophages IBD onset and progression need be added.
6. Some discussion on the available therapeutic strategies targeting macrophage polarization
will be appreciated.
7. Figure 4 is pack with panel, it is advised to separate into a new figure.
Please review this paper.
user
Title: Macrophage polarization toward M1 phenotype in T cell transfer colitis model
Version: 1 Date: 28 Oct 2023
Reviewer's report:
The manuscript from Ebihara et al. aimed to investigate the macrophage function in a T
cell transfer colitis murine model, and found that the macrophages polarized toward the M1
phenotype in the colonic LP. The polarized macrophages also activated the colitogenic T cells,
which may contribute to disease progression and could be a potential therapeutic target. This
manuscript is well-written, and elucidated the unappreciated role of macrophages in a T cell
transfer colitis model. This study has the potential to lead to several subsequent projects in
order to study more mechanisms and therapeutic measures. However, significant revisions are
suggested to improve the quality of the manuscript.
1. The presentation of flow cytometry data needs to be improved significantly. In figure 3,
instead of using exact numbers to calculate macrophages, inflammatory monocytes, neutrophils,
and T cells. The author should consider using their ratio to CD45+ cells (such as number of
macrophages/number of CD45+ cells), that would be a more objective way to present.
2. The gating strategies among figure 3 (F-I) and figure 3 (J-M) are different, the author should
standardized the gating, or explain why using different strategies. In order to quantify the
intracellular cytokines, it is advised to use Mean, or Median, Fluorescence Intensity (MFI).
3. Since CD11c is also a marker for DCs, and DCs are considered as innate immune cells,
the author may want to discuss how they differentiate DCs and M1 macrophages from CD11c
positive cells.
4. Regarding the histological score of colitis, it is better to list the individual score for epithelial
hyperplasia, goblet cell and crypt loss and inflammatory infiltration. This will help understand
which of the three criteria is the main contributer to the increased histological score. Arrows are
suggested to mark the different histological changes in panels of F, G and H from figure 1.
5. In spite the widely recognized role of T cells in IBD, more discussions on the role of
macrophages IBD onset and progression need be added.
6. Some discussion on the available therapeutic strategies targeting macrophage polarization
will be appreciated.
7. Figure 4 is pack with panel, it is advised to separate into a new figure.
assistant