Reviewer's report
Title:Murine CD4+CD25- cells activated in vitro with PMA/ionomycin and
anti-CD3 acquire regulatory function and ameliorate experimental colitis in vivo
Version: 1Date:5 October 2012
Reviewer: Franco Scaldaferri
Reviewer's report:
Potentially interesting paper, assessing the T regulatory capacity of subset of T
cells manipulated in vitro in the experimental murine CD45RB transfer model of
colitis.
Well written, clear however it displays several major limitations.
Authors state that murine cd4+cd25- cells activated in vitro with pma/ionomycin
and anti-cd3 acquire regulatory function.
However, even if they are associated to a “clinical amelioration” of colitis in mice,
the causality could not be related so strictly to those “supposed” transferred
regulatory T cells.
Those cells, in fact, have not been fully characterized in vitro, in simplified
system. Do they display regulatory properties in vitro? are they able to suppress
proliferation of activated T cells in a dose dependent manner? Do they inhibit T
cells cytokines production upon activation?
Do they stay alive in mice following the transfer?
Furthermore, authors state that it is the treatment with pma/ionomycin, by
modulating ion channel function, that enhances the expression of cd25 on murine
cd4+cd25- cells. A control group of mice receiving cd4+cd25- cells activated only
with anti-CD3 and then cultured in IL2 supplemented medium should also be
included in the paper.
Finally introduction should contain more information about stimulation of T cells
with (PMA)/ionomycin and anti-CD3.
Aim of the study should be stated instead of anticipating, already in the
introduction, results of the study and analysis.
In the discussion session more room should be dedicated to compare
pma/ionomycin and anti-cd3-activated murine cd4+cd25- cells to other in vitro
generated regulatory T cells. Limits of the paper should also be better presented.
Level of interest: An article of importance in its field
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:Murine CD4+CD25- cells activated in vitro with PMA/ionomycin and
anti-CD3 acquire regulatory function and ameliorate experimental colitis in vivo
Version: 1Date:5 October 2012
Reviewer: Franco Scaldaferri
Reviewer's report:
Potentially interesting paper, assessing the T regulatory capacity of subset of T
cells manipulated in vitro in the experimental murine CD45RB transfer model of
colitis.
Well written, clear however it displays several major limitations.
Authors state that murine cd4+cd25- cells activated in vitro with pma/ionomycin
and anti-cd3 acquire regulatory function.
However, even if they are associated to a “clinical amelioration” of colitis in mice,
the causality could not be related so strictly to those “supposed” transferred
regulatory T cells.
Those cells, in fact, have not been fully characterized in vitro, in simplified
system. Do they display regulatory properties in vitro? are they able to suppress
proliferation of activated T cells in a dose dependent manner? Do they inhibit T
cells cytokines production upon activation?
Do they stay alive in mice following the transfer?
Furthermore, authors state that it is the treatment with pma/ionomycin, by
modulating ion channel function, that enhances the expression of cd25 on murine
cd4+cd25- cells. A control group of mice receiving cd4+cd25- cells activated only
with anti-CD3 and then cultured in IL2 supplemented medium should also be
included in the paper.
Finally introduction should contain more information about stimulation of T cells
with (PMA)/ionomycin and anti-CD3.
Aim of the study should be stated instead of anticipating, already in the
introduction, results of the study and analysis.
In the discussion session more room should be dedicated to compare
pma/ionomycin and anti-cd3-activated murine cd4+cd25- cells to other in vitro
generated regulatory T cells. Limits of the paper should also be better presented.
Level of interest: An article of importance in its field
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:Murine CD4+CD25- cells activated in vitro with PMA/ionomycin and
anti-CD3 acquire regulatory function and ameliorate experimental colitis in vivo
Version: 1Date:5 October 2012
Reviewer: Franco Scaldaferri
Reviewer's report:
Potentially interesting paper, assessing the T regulatory capacity of subset of T
cells manipulated in vitro in the experimental murine CD45RB transfer model of
colitis.
Well written, clear however it displays several major limitations.
Authors state that murine cd4+cd25- cells activated in vitro with pma/ionomycin
and anti-cd3 acquire regulatory function.
However, even if they are associated to a “clinical amelioration” of colitis in mice,
the causality could not be related so strictly to those “supposed” transferred
regulatory T cells.
Those cells, in fact, have not been fully characterized in vitro, in simplified
system. Do they display regulatory properties in vitro? are they able to suppress
proliferation of activated T cells in a dose dependent manner? Do they inhibit T
cells cytokines production upon activation?
Do they stay alive in mice following the transfer?
Furthermore, authors state that it is the treatment with pma/ionomycin, by
modulating ion channel function, that enhances the expression of cd25 on murine
cd4+cd25- cells. A control group of mice receiving cd4+cd25- cells activated only
with anti-CD3 and then cultured in IL2 supplemented medium should also be
included in the paper.
Finally introduction should contain more information about stimulation of T cells
with (PMA)/ionomycin and anti-CD3.
Aim of the study should be stated instead of anticipating, already in the
introduction, results of the study and analysis.
In the discussion session more room should be dedicated to compare
pma/ionomycin and anti-cd3-activated murine cd4+cd25- cells to other in vitro
generated regulatory T cells. Limits of the paper should also be better presented.
Level of interest: An article of importance in its field
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'
assistant