Reviewer's report 
 Transfection of IL-10 expression vectors into endothelial cultures attenuates alpha 4 beta Title: 
7-dependent lymphocyte adhesion mediated by MAdCAM-1 
Authors: 
Makoto Sasaki ( msasak@lsuhsc.edu )
Paul Jordan ( pjorda1@lsuhsc.edu )
Jeff Houghton ( jhough@lsuhsc.edu )
Xianmin Meng ( mengderm@yahoo.com )
Makoto Itoh ( mitoh@med.nagoya-cu.ac.jp )
Takashi Joh ( tjoh@med.nagoya-cu.ac.jp )
J. S. Alexander ( jalexa@lsuhsc.edu )
 1  24 Dec 2002 Version: Date: 
 Dr Soichiro Miura Reviewer: 
 A paper of considerable general medical or scientific interest Level of interest: 
 Accept after discretionary revisions Advice on publication: 
This is an interesting report in which authors suggested that gene therapy of the gut microvasculature 
with IL-10 vectors significantly attenuated the TNF-alpha-induced lymphocyte adhesion to endothelial 
cells as well as TNF-alpha-induced expression of MAdCAM-1. However, I have several concerns as 
follows; 
1. In this manuscript, the authors describe SVEC4-10 cell line as lymphatic endothelial cells. I suppose 
that this is a venular endothelial cell line, not a lymphatic-endothelium derived cell line? Since biological 
nature of lymphatic endothelial cells are largely different from that of venular endothelial cells, the 
vascular origin of this cell line should be carefully presented. 
2. In this paper, there is no evidence that TNF-alpha-stimulated lymphocyte adhesion was due to the enhanced MAdCAM-1 expression. Because TNF-alpha may also upregulate other adhesion molecules (i.e. ICAM-1, VCAM-1 etc.), and because the TK-1 adhesion is only 50% MAdCAM-1 dependent, the 
authors should clarify the effect of IL-10 transfection on other adhesion molecules, and their roles in 
enhanced lymphocyte adhesion. 3.It is intriguing that IL-10 transfection did not affect the basal levels of MAdCAM-1 expression as well as lymphocyte adhesion to the endothelial cells. This means that there is a significant difference in 
regulation of MAdCAM-1 expression between basal and stimulated conditions. Please explain more in 
detail about the possible reason for this in the Discussion. 
Minor Comments: 
1. How IL-10 transfection affect the morphology, proliferation or permeability of this cell line? These 
informations are important to the reader to consider clinical application. 2. Abstract line 13, TNF-alpha induced -1 expression. 
Competing interests: 
None declared. 
Reviewer's report 
 Transfection of IL-10 expression vectors into endothelial cultures attenuates alpha 4 beta Title: 
7-dependent lymphocyte adhesion mediated by MAdCAM-1 
Authors: 
Makoto Sasaki ( msasak@lsuhsc.edu )
Paul Jordan ( pjorda1@lsuhsc.edu )
Jeff Houghton ( jhough@lsuhsc.edu )
Xianmin Meng ( mengderm@yahoo.com )
Makoto Itoh ( mitoh@med.nagoya-cu.ac.jp )
Takashi Joh ( tjoh@med.nagoya-cu.ac.jp )
J. S. Alexander ( jalexa@lsuhsc.edu )
 1  24 Dec 2002 Version: Date: 
 Dr Soichiro Miura Reviewer: 
 A paper of considerable general medical or scientific interest Level of interest: 
 Accept after discretionary revisions Advice on publication: 
This is an interesting report in which authors suggested that gene therapy of the gut microvasculature 
with IL-10 vectors significantly attenuated the TNF-alpha-induced lymphocyte adhesion to endothelial 
cells as well as TNF-alpha-induced expression of MAdCAM-1. However, I have several concerns as 
follows; 
1. In this manuscript, the authors describe SVEC4-10 cell line as lymphatic endothelial cells. I suppose 
that this is a venular endothelial cell line, not a lymphatic-endothelium derived cell line? Since biological 
nature of lymphatic endothelial cells are largely different from that of venular endothelial cells, the 
vascular origin of this cell line should be carefully presented. 
2. In this paper, there is no evidence that TNF-alpha-stimulated lymphocyte adhesion was due to the enhanced MAdCAM-1 expression. Because TNF-alpha may also upregulate other adhesion molecules (i.e. ICAM-1, VCAM-1 etc.), and because the TK-1 adhesion is only 50% MAdCAM-1 dependent, the 
authors should clarify the effect of IL-10 transfection on other adhesion molecules, and their roles in 
enhanced lymphocyte adhesion. 3.It is intriguing that IL-10 transfection did not affect the basal levels of MAdCAM-1 expression as well as lymphocyte adhesion to the endothelial cells. This means that there is a significant difference in 
regulation of MAdCAM-1 expression between basal and stimulated conditions. Please explain more in 
detail about the possible reason for this in the Discussion. 
Minor Comments: 
1. How IL-10 transfection affect the morphology, proliferation or permeability of this cell line? These 
informations are important to the reader to consider clinical application. 2. Abstract line 13, TNF-alpha induced -1 expression. 
Competing interests: 
None declared. 
Please review this paper.
user
Reviewer's report 
 Transfection of IL-10 expression vectors into endothelial cultures attenuates alpha 4 beta Title: 
7-dependent lymphocyte adhesion mediated by MAdCAM-1 
Authors: 
Makoto Sasaki ( msasak@lsuhsc.edu )
Paul Jordan ( pjorda1@lsuhsc.edu )
Jeff Houghton ( jhough@lsuhsc.edu )
Xianmin Meng ( mengderm@yahoo.com )
Makoto Itoh ( mitoh@med.nagoya-cu.ac.jp )
Takashi Joh ( tjoh@med.nagoya-cu.ac.jp )
J. S. Alexander ( jalexa@lsuhsc.edu )
 1  24 Dec 2002 Version: Date: 
 Dr Soichiro Miura Reviewer: 
 A paper of considerable general medical or scientific interest Level of interest: 
 Accept after discretionary revisions Advice on publication: 
This is an interesting report in which authors suggested that gene therapy of the gut microvasculature 
with IL-10 vectors significantly attenuated the TNF-alpha-induced lymphocyte adhesion to endothelial 
cells as well as TNF-alpha-induced expression of MAdCAM-1. However, I have several concerns as 
follows; 
1. In this manuscript, the authors describe SVEC4-10 cell line as lymphatic endothelial cells. I suppose 
that this is a venular endothelial cell line, not a lymphatic-endothelium derived cell line? Since biological 
nature of lymphatic endothelial cells are largely different from that of venular endothelial cells, the 
vascular origin of this cell line should be carefully presented. 
2. In this paper, there is no evidence that TNF-alpha-stimulated lymphocyte adhesion was due to the enhanced MAdCAM-1 expression. Because TNF-alpha may also upregulate other adhesion molecules (i.e. ICAM-1, VCAM-1 etc.), and because the TK-1 adhesion is only 50% MAdCAM-1 dependent, the 
authors should clarify the effect of IL-10 transfection on other adhesion molecules, and their roles in 
enhanced lymphocyte adhesion. 3.It is intriguing that IL-10 transfection did not affect the basal levels of MAdCAM-1 expression as well as lymphocyte adhesion to the endothelial cells. This means that there is a significant difference in 
regulation of MAdCAM-1 expression between basal and stimulated conditions. Please explain more in 
detail about the possible reason for this in the Discussion. 
Minor Comments: 
1. How IL-10 transfection affect the morphology, proliferation or permeability of this cell line? These 
informations are important to the reader to consider clinical application. 2. Abstract line 13, TNF-alpha induced -1 expression. 
Competing interests: 
None declared. 
assistant