Reviewer's report
Title:Lamina propria macrophage phenotypes in relation to Escherichia coli in
Crohn's disease
Version:2Date:18 February 2015
Reviewer: Nobuhiko Kamada
Reviewer's report:
In this manuscript, Elliott et al investigated the phenotypes of E. coli-laden and E.
coli-unladen macrophages isolated from the intestinal lamina propria of IBD
patients and healthy controls. E. coli-laden macrophages are commonly found in
patients with Crohn’s disease (CD) compared to patients with ulcerative colitis
(UC). No E. coli-laden macrophages were found in healthy controls. Next, the
authors assessed the phenotypes of these macrophages isolated by laser
capture microdissection. Expression levels of cytokines (TNF, IL-23, IL-6, and
IL-8) and inflammatory mediators (iNOS, COX-2) were higher in CD patients than
in control subjects. In CD patients, the E. coli-laden macrophages were found to
express lower levels of pro-inflammatory cytokines (TNF, IL-23, IL-6, IL-8)
compared to the E. coli-unladen macrophages. In contrast, the expression of an
anti-inflammatory cytokine, IL-10, was greater in the E. coli-laden macrophages
than in the E. coli-unladen macrophages. However, in the tissue not affected by
inflammation, the E. coli-laden macrophages express higher levels of
pro-inflammatory cytokines compared to the E. coli-unladen macrophages.
Although the phenotypic characterization of E. coli-laden macrophages in CD
patients is of interest, there are still significant concerns that need to be
addressed to strengthen the main claim of this manuscript. Specific comments
are below:
Major:
1. The authors identified the E. coli-laden macrophages by immunostaining E.
coli. However, the specificity of this method is unclear. To strengthen their claim,
the authors should consider using another method to confirm the presence of E.
coli in the macrophages. For example, the authors could confirm the presence of
E. coli in isolated macrophages by PCR (16S rRNA specific for E. coli).
2. Why Fig. 4B and G did not have a UC patient group?
Discretionary:
1. Similar to comment #1, it would be informative to test whether these
macrophages contain bacteria other than E. coli. Is E. coli the only bacterium
present in the macrophages? Are there any other bacteria present in the E.
coli-unladen macrophage (using universal bacterial 16S rRNA primers)?
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Reviewer's report
Title:Lamina propria macrophage phenotypes in relation to Escherichia coli in
Crohn's disease
Version:2Date:18 February 2015
Reviewer: Nobuhiko Kamada
Reviewer's report:
In this manuscript, Elliott et al investigated the phenotypes of E. coli-laden and E.
coli-unladen macrophages isolated from the intestinal lamina propria of IBD
patients and healthy controls. E. coli-laden macrophages are commonly found in
patients with Crohn’s disease (CD) compared to patients with ulcerative colitis
(UC). No E. coli-laden macrophages were found in healthy controls. Next, the
authors assessed the phenotypes of these macrophages isolated by laser
capture microdissection. Expression levels of cytokines (TNF, IL-23, IL-6, and
IL-8) and inflammatory mediators (iNOS, COX-2) were higher in CD patients than
in control subjects. In CD patients, the E. coli-laden macrophages were found to
express lower levels of pro-inflammatory cytokines (TNF, IL-23, IL-6, IL-8)
compared to the E. coli-unladen macrophages. In contrast, the expression of an
anti-inflammatory cytokine, IL-10, was greater in the E. coli-laden macrophages
than in the E. coli-unladen macrophages. However, in the tissue not affected by
inflammation, the E. coli-laden macrophages express higher levels of
pro-inflammatory cytokines compared to the E. coli-unladen macrophages.
Although the phenotypic characterization of E. coli-laden macrophages in CD
patients is of interest, there are still significant concerns that need to be
addressed to strengthen the main claim of this manuscript. Specific comments
are below:
Major:
1. The authors identified the E. coli-laden macrophages by immunostaining E.
coli. However, the specificity of this method is unclear. To strengthen their claim,
the authors should consider using another method to confirm the presence of E.
coli in the macrophages. For example, the authors could confirm the presence of
E. coli in isolated macrophages by PCR (16S rRNA specific for E. coli).
2. Why Fig. 4B and G did not have a UC patient group?
Discretionary:
1. Similar to comment #1, it would be informative to test whether these
macrophages contain bacteria other than E. coli. Is E. coli the only bacterium
present in the macrophages? Are there any other bacteria present in the E.
coli-unladen macrophage (using universal bacterial 16S rRNA primers)?
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Please review this paper.
user
Reviewer's report
Title:Lamina propria macrophage phenotypes in relation to Escherichia coli in
Crohn's disease
Version:2Date:18 February 2015
Reviewer: Nobuhiko Kamada
Reviewer's report:
In this manuscript, Elliott et al investigated the phenotypes of E. coli-laden and E.
coli-unladen macrophages isolated from the intestinal lamina propria of IBD
patients and healthy controls. E. coli-laden macrophages are commonly found in
patients with Crohn’s disease (CD) compared to patients with ulcerative colitis
(UC). No E. coli-laden macrophages were found in healthy controls. Next, the
authors assessed the phenotypes of these macrophages isolated by laser
capture microdissection. Expression levels of cytokines (TNF, IL-23, IL-6, and
IL-8) and inflammatory mediators (iNOS, COX-2) were higher in CD patients than
in control subjects. In CD patients, the E. coli-laden macrophages were found to
express lower levels of pro-inflammatory cytokines (TNF, IL-23, IL-6, IL-8)
compared to the E. coli-unladen macrophages. In contrast, the expression of an
anti-inflammatory cytokine, IL-10, was greater in the E. coli-laden macrophages
than in the E. coli-unladen macrophages. However, in the tissue not affected by
inflammation, the E. coli-laden macrophages express higher levels of
pro-inflammatory cytokines compared to the E. coli-unladen macrophages.
Although the phenotypic characterization of E. coli-laden macrophages in CD
patients is of interest, there are still significant concerns that need to be
addressed to strengthen the main claim of this manuscript. Specific comments
are below:
Major:
1. The authors identified the E. coli-laden macrophages by immunostaining E.
coli. However, the specificity of this method is unclear. To strengthen their claim,
the authors should consider using another method to confirm the presence of E.
coli in the macrophages. For example, the authors could confirm the presence of
E. coli in isolated macrophages by PCR (16S rRNA specific for E. coli).
2. Why Fig. 4B and G did not have a UC patient group?
Discretionary:
1. Similar to comment #1, it would be informative to test whether these
macrophages contain bacteria other than E. coli. Is E. coli the only bacterium
present in the macrophages? Are there any other bacteria present in the E.
coli-unladen macrophage (using universal bacterial 16S rRNA primers)?
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
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