Reviewer's report
Title:Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter
rodentium-induced colitis.
Version: 1Date:7 February 2013
Reviewer: Chihiro Sasakawa
Reviewer's report:
Assi et al. describe that integrin-linked kinase (ILK)-knockout (ko) mice shows
reduced inflammation and attenuated hyperplastic cryptal response to
Citrobacter rodentium infection. In the authors’ recent studies (2008 Gut and
2011 BMC Immunology), they have shown that ILK-ko mice also exhibited less
wasting disease or attenuated carcinogenesis in response to dextran sodium
sulfate (DSS) treatment. Their findings are intriguing, since ILK potentiates
colonic inflammation through perhaps indirect effect on chemokine expression,
epithelial cell proliferation, and extracellular matrix protein expression induced by
both DSS treatment and C. rodentium infection. However, it is not clear that the
changes are caused by bacterial infection or ILK-loss. Though the present study
finds potential significance, additional evidence is required to draw any
conclusion.
Major comments;
1) The authors showed the reduction of cell proliferation (Ki-67 positive cell
number), crypt heights and the level of cyclinD1 at both 6 and 15 days post
infection in the ILK-ko mice (figure 8, 9). However, they have already reported
that the reduction of proliferating cell number, crypt heights, and cyclinD1
expression level in non-infected ILK-ko mice compared to WT mice (Gut 2008,
figure 2C, D, G, E, H). It is not clear the results that are shown here are due to
the ILK-loss per se or bacterial infection. They should compare the infected colon
with non-infected control in both WT and ILK-ko mice.
2) In the GUT paper (2008), the ILK-KO mice colon architecture is different from
that of wild type. They did not show non-infected control in both WT and ILK-ko
mice throughout the paper. Therefore it is difficult to judge the results. They
should show non-infected control in both WT and ILK-KO mice as following
figures; figure 2E, 3C, 9, 10.
3) The authors showed that bacteria cannot migrate downwards along the lateral
aspect of crypts in ILK-ko mice colon (figure 6D) in compared with WT mice
colon (figure6C). However the difference is not evident in these figures, because
ILK-ko mice colon crypts are cut latitudinally in figure 6D, while WT colon crypts
are cut longitudinally.
4) The authors said the bacterial colonization is not change between WT and
ILK-ko mice.
However, LPS and TIR positive signal shows big difference between these two
groups. Please give some comments.
5) The authors said that the colon stem cell compartment is stained positive for
cyclinD1 in figure 9. CyclinD1 is not a proper stem cell marker of colon. They
need to stain with the proper stem cell marker protein such as Lgr5.
Minor comments:
1) Figure 5A is not clear. Therefore it is difficult to judge the results mentioned for
this experiment.
2) C. rodentium and CyclinD are not properly labeled.
3) Figures are not properly labeled.
4) Many critical literatures through the text are missing.
5) Many references need to put published year.
Reviewer's report
Title:Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter
rodentium-induced colitis.
Version: 1Date:7 February 2013
Reviewer: Chihiro Sasakawa
Reviewer's report:
Assi et al. describe that integrin-linked kinase (ILK)-knockout (ko) mice shows
reduced inflammation and attenuated hyperplastic cryptal response to
Citrobacter rodentium infection. In the authors’ recent studies (2008 Gut and
2011 BMC Immunology), they have shown that ILK-ko mice also exhibited less
wasting disease or attenuated carcinogenesis in response to dextran sodium
sulfate (DSS) treatment. Their findings are intriguing, since ILK potentiates
colonic inflammation through perhaps indirect effect on chemokine expression,
epithelial cell proliferation, and extracellular matrix protein expression induced by
both DSS treatment and C. rodentium infection. However, it is not clear that the
changes are caused by bacterial infection or ILK-loss. Though the present study
finds potential significance, additional evidence is required to draw any
conclusion.
Major comments;
1) The authors showed the reduction of cell proliferation (Ki-67 positive cell
number), crypt heights and the level of cyclinD1 at both 6 and 15 days post
infection in the ILK-ko mice (figure 8, 9). However, they have already reported
that the reduction of proliferating cell number, crypt heights, and cyclinD1
expression level in non-infected ILK-ko mice compared to WT mice (Gut 2008,
figure 2C, D, G, E, H). It is not clear the results that are shown here are due to
the ILK-loss per se or bacterial infection. They should compare the infected colon
with non-infected control in both WT and ILK-ko mice.
2) In the GUT paper (2008), the ILK-KO mice colon architecture is different from
that of wild type. They did not show non-infected control in both WT and ILK-ko
mice throughout the paper. Therefore it is difficult to judge the results. They
should show non-infected control in both WT and ILK-KO mice as following
figures; figure 2E, 3C, 9, 10.
3) The authors showed that bacteria cannot migrate downwards along the lateral
aspect of crypts in ILK-ko mice colon (figure 6D) in compared with WT mice
colon (figure6C). However the difference is not evident in these figures, because
ILK-ko mice colon crypts are cut latitudinally in figure 6D, while WT colon crypts
are cut longitudinally.
4) The authors said the bacterial colonization is not change between WT and
ILK-ko mice.
However, LPS and TIR positive signal shows big difference between these two
groups. Please give some comments.
5) The authors said that the colon stem cell compartment is stained positive for
cyclinD1 in figure 9. CyclinD1 is not a proper stem cell marker of colon. They
need to stain with the proper stem cell marker protein such as Lgr5.
Minor comments:
1) Figure 5A is not clear. Therefore it is difficult to judge the results mentioned for
this experiment.
2) C. rodentium and CyclinD are not properly labeled.
3) Figures are not properly labeled.
4) Many critical literatures through the text are missing.
5) Many references need to put published year.
Please review this paper.
user
Reviewer's report
Title:Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter
rodentium-induced colitis.
Version: 1Date:7 February 2013
Reviewer: Chihiro Sasakawa
Reviewer's report:
Assi et al. describe that integrin-linked kinase (ILK)-knockout (ko) mice shows
reduced inflammation and attenuated hyperplastic cryptal response to
Citrobacter rodentium infection. In the authors’ recent studies (2008 Gut and
2011 BMC Immunology), they have shown that ILK-ko mice also exhibited less
wasting disease or attenuated carcinogenesis in response to dextran sodium
sulfate (DSS) treatment. Their findings are intriguing, since ILK potentiates
colonic inflammation through perhaps indirect effect on chemokine expression,
epithelial cell proliferation, and extracellular matrix protein expression induced by
both DSS treatment and C. rodentium infection. However, it is not clear that the
changes are caused by bacterial infection or ILK-loss. Though the present study
finds potential significance, additional evidence is required to draw any
conclusion.
Major comments;
1) The authors showed the reduction of cell proliferation (Ki-67 positive cell
number), crypt heights and the level of cyclinD1 at both 6 and 15 days post
infection in the ILK-ko mice (figure 8, 9). However, they have already reported
that the reduction of proliferating cell number, crypt heights, and cyclinD1
expression level in non-infected ILK-ko mice compared to WT mice (Gut 2008,
figure 2C, D, G, E, H). It is not clear the results that are shown here are due to
the ILK-loss per se or bacterial infection. They should compare the infected colon
with non-infected control in both WT and ILK-ko mice.
2) In the GUT paper (2008), the ILK-KO mice colon architecture is different from
that of wild type. They did not show non-infected control in both WT and ILK-ko
mice throughout the paper. Therefore it is difficult to judge the results. They
should show non-infected control in both WT and ILK-KO mice as following
figures; figure 2E, 3C, 9, 10.
3) The authors showed that bacteria cannot migrate downwards along the lateral
aspect of crypts in ILK-ko mice colon (figure 6D) in compared with WT mice
colon (figure6C). However the difference is not evident in these figures, because
ILK-ko mice colon crypts are cut latitudinally in figure 6D, while WT colon crypts
are cut longitudinally.
4) The authors said the bacterial colonization is not change between WT and
ILK-ko mice.
However, LPS and TIR positive signal shows big difference between these two
groups. Please give some comments.
5) The authors said that the colon stem cell compartment is stained positive for
cyclinD1 in figure 9. CyclinD1 is not a proper stem cell marker of colon. They
need to stain with the proper stem cell marker protein such as Lgr5.
Minor comments:
1) Figure 5A is not clear. Therefore it is difficult to judge the results mentioned for
this experiment.
2) C. rodentium and CyclinD are not properly labeled.
3) Figures are not properly labeled.
4) Many critical literatures through the text are missing.
5) Many references need to put published year.
assistant