Reviewer's report
Title:Small intestinal mucosa expression of putative chaperone fls485
Version: 1Date:8 July 2009
Reviewer: Mark Musch
Reviewer's report:
The manuscript of Reinartz et al investigates the expression of a chaperone
protein fls485 in intestinal epithelia. This gene product has received little
attention, however, it possesses many domains that suggest it could be an
important regulator of cell growth and or differentiation. An important facet of the
studies is the generation of antibodies that allow assessment of protein
expression. The authors determine that this gene product is expressed in
intestinal epithelial cells in both crypt and villus enterocytes, but the expression is
higher in the villus cells, consistent with a potential role in differentiation. Fls485
expression was also found in some cells of the lamina propria, supporting this its
expression is not confined to epithelial cells. The investigators also demonstrate
expression in a number of cultured intestinal epithelial cell lines. Fls485 is
determined to be a cytosolic protein and the expression is decreased in celiac
disease. The studies are potentially interesting and fls485 has the probability of
being an important regulatory protein in the epithelial cells, however, at present
the impact of the studies is limited since a functional role has not been
established.
Major Compulsory Revisions
(1)The investigators have the opportunity to determine in fls485 may have a role
in differentiation using an established cultured in vitro cell line, Caco2. This cell
line differentiates post-confluence in culture. Expression of villin,
cysteine-sensitive alkaline phosphatase, and sucrase increase after Caco2 cells
come to contact inhibition of growth. While it may be presumptive to state that
this is precisely representative of maturation of villus small intestinal enterocytes,
it is a model that is well accepted. It is stated that this cell line expresses fls485
and the investigators have the expertise to determine if fls485 expression
increases as these cells mature. This investigation should also include studies of
the maturation to determine if one or more of the differentiated characteristics are
regulated by fls485. The investigators could extend these studies using RNA
silencing technologies to determine whether fls485 is necessary for maturation or
simply expression parallels maturation of other markers.
(2) Another point that could be presented more clearly is the reason for
investigations of celiac disease and not other intestinal pathologies. Fls485
expression might be anticipated to be up or downregulated by a number of
conditions such as bacterial infections by Salmonella or viral infection by
rotavirus. These two conditions may be difficult to obtain, however, through the
connection to the Pathology department, the investigators may be able to obtain
sections of inflammatory bowel diseases, both pediatric and adult. It is
appreciated that immunohistochemistry is a poorly quantitative technology
however, to obtain fresh tissues for Western blotting and better quantitation
would be excellent, but perhaps beyond the scope of the present studies.
Another condition that might regulate fls485 expression is colon cancer where
the importance of other chaperone proteins is being established as biomarkers
as well as in the pathogenesis of the disease. Sections from a Pathology bank
could well provide important information that allow further investigation of the
roles of this protein.
Minor Essential Revisions
(1) For the studies of intestine, it would be good to also include a longitudinal
study of fls485 expression. Sections of stomach, duodenum, jejunum, ileum,
cecum, proximal and distal colon are readily obtained from animal models and
perhaps the investigators have access to a pathology bank of preserved tissues.
For these sections, it would be interesting and potentially important to establish
which epithelial cells in each segment express fls485. By itself this survey is not
sufficient for publication, but it would add to the initial investigations of this gene
product.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: Yes, and I have assessed the statistics in my report.
Declaration of competing interests:
I declare that I have no competing interests
Reviewer's report
Title:Small intestinal mucosa expression of putative chaperone fls485
Version: 1Date:8 July 2009
Reviewer: Mark Musch
Reviewer's report:
The manuscript of Reinartz et al investigates the expression of a chaperone
protein fls485 in intestinal epithelia. This gene product has received little
attention, however, it possesses many domains that suggest it could be an
important regulator of cell growth and or differentiation. An important facet of the
studies is the generation of antibodies that allow assessment of protein
expression. The authors determine that this gene product is expressed in
intestinal epithelial cells in both crypt and villus enterocytes, but the expression is
higher in the villus cells, consistent with a potential role in differentiation. Fls485
expression was also found in some cells of the lamina propria, supporting this its
expression is not confined to epithelial cells. The investigators also demonstrate
expression in a number of cultured intestinal epithelial cell lines. Fls485 is
determined to be a cytosolic protein and the expression is decreased in celiac
disease. The studies are potentially interesting and fls485 has the probability of
being an important regulatory protein in the epithelial cells, however, at present
the impact of the studies is limited since a functional role has not been
established.
Major Compulsory Revisions
(1)The investigators have the opportunity to determine in fls485 may have a role
in differentiation using an established cultured in vitro cell line, Caco2. This cell
line differentiates post-confluence in culture. Expression of villin,
cysteine-sensitive alkaline phosphatase, and sucrase increase after Caco2 cells
come to contact inhibition of growth. While it may be presumptive to state that
this is precisely representative of maturation of villus small intestinal enterocytes,
it is a model that is well accepted. It is stated that this cell line expresses fls485
and the investigators have the expertise to determine if fls485 expression
increases as these cells mature. This investigation should also include studies of
the maturation to determine if one or more of the differentiated characteristics are
regulated by fls485. The investigators could extend these studies using RNA
silencing technologies to determine whether fls485 is necessary for maturation or
simply expression parallels maturation of other markers.
(2) Another point that could be presented more clearly is the reason for
investigations of celiac disease and not other intestinal pathologies. Fls485
expression might be anticipated to be up or downregulated by a number of
conditions such as bacterial infections by Salmonella or viral infection by
rotavirus. These two conditions may be difficult to obtain, however, through the
connection to the Pathology department, the investigators may be able to obtain
sections of inflammatory bowel diseases, both pediatric and adult. It is
appreciated that immunohistochemistry is a poorly quantitative technology
however, to obtain fresh tissues for Western blotting and better quantitation
would be excellent, but perhaps beyond the scope of the present studies.
Another condition that might regulate fls485 expression is colon cancer where
the importance of other chaperone proteins is being established as biomarkers
as well as in the pathogenesis of the disease. Sections from a Pathology bank
could well provide important information that allow further investigation of the
roles of this protein.
Minor Essential Revisions
(1) For the studies of intestine, it would be good to also include a longitudinal
study of fls485 expression. Sections of stomach, duodenum, jejunum, ileum,
cecum, proximal and distal colon are readily obtained from animal models and
perhaps the investigators have access to a pathology bank of preserved tissues.
For these sections, it would be interesting and potentially important to establish
which epithelial cells in each segment express fls485. By itself this survey is not
sufficient for publication, but it would add to the initial investigations of this gene
product.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: Yes, and I have assessed the statistics in my report.
Declaration of competing interests:
I declare that I have no competing interests
Please review this paper.
user
Reviewer's report
Title:Small intestinal mucosa expression of putative chaperone fls485
Version: 1Date:8 July 2009
Reviewer: Mark Musch
Reviewer's report:
The manuscript of Reinartz et al investigates the expression of a chaperone
protein fls485 in intestinal epithelia. This gene product has received little
attention, however, it possesses many domains that suggest it could be an
important regulator of cell growth and or differentiation. An important facet of the
studies is the generation of antibodies that allow assessment of protein
expression. The authors determine that this gene product is expressed in
intestinal epithelial cells in both crypt and villus enterocytes, but the expression is
higher in the villus cells, consistent with a potential role in differentiation. Fls485
expression was also found in some cells of the lamina propria, supporting this its
expression is not confined to epithelial cells. The investigators also demonstrate
expression in a number of cultured intestinal epithelial cell lines. Fls485 is
determined to be a cytosolic protein and the expression is decreased in celiac
disease. The studies are potentially interesting and fls485 has the probability of
being an important regulatory protein in the epithelial cells, however, at present
the impact of the studies is limited since a functional role has not been
established.
Major Compulsory Revisions
(1)The investigators have the opportunity to determine in fls485 may have a role
in differentiation using an established cultured in vitro cell line, Caco2. This cell
line differentiates post-confluence in culture. Expression of villin,
cysteine-sensitive alkaline phosphatase, and sucrase increase after Caco2 cells
come to contact inhibition of growth. While it may be presumptive to state that
this is precisely representative of maturation of villus small intestinal enterocytes,
it is a model that is well accepted. It is stated that this cell line expresses fls485
and the investigators have the expertise to determine if fls485 expression
increases as these cells mature. This investigation should also include studies of
the maturation to determine if one or more of the differentiated characteristics are
regulated by fls485. The investigators could extend these studies using RNA
silencing technologies to determine whether fls485 is necessary for maturation or
simply expression parallels maturation of other markers.
(2) Another point that could be presented more clearly is the reason for
investigations of celiac disease and not other intestinal pathologies. Fls485
expression might be anticipated to be up or downregulated by a number of
conditions such as bacterial infections by Salmonella or viral infection by
rotavirus. These two conditions may be difficult to obtain, however, through the
connection to the Pathology department, the investigators may be able to obtain
sections of inflammatory bowel diseases, both pediatric and adult. It is
appreciated that immunohistochemistry is a poorly quantitative technology
however, to obtain fresh tissues for Western blotting and better quantitation
would be excellent, but perhaps beyond the scope of the present studies.
Another condition that might regulate fls485 expression is colon cancer where
the importance of other chaperone proteins is being established as biomarkers
as well as in the pathogenesis of the disease. Sections from a Pathology bank
could well provide important information that allow further investigation of the
roles of this protein.
Minor Essential Revisions
(1) For the studies of intestine, it would be good to also include a longitudinal
study of fls485 expression. Sections of stomach, duodenum, jejunum, ileum,
cecum, proximal and distal colon are readily obtained from animal models and
perhaps the investigators have access to a pathology bank of preserved tissues.
For these sections, it would be interesting and potentially important to establish
which epithelial cells in each segment express fls485. By itself this survey is not
sufficient for publication, but it would add to the initial investigations of this gene
product.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: Yes, and I have assessed the statistics in my report.
Declaration of competing interests:
I declare that I have no competing interests
assistant