Comparison of E-selectin expression at mRNA and protein levels in murine models of inflammation.
BACKGROUND
Drug targeting to activated endothelial cells via E-selectin is currently being explored as a new approach to treat chronic inflammatory disorders. This approach uses E-selectin directed antibodies as carrier molecules to selectively deliver anti-inflammatory drugs into activated endothelial cells, thereby theoretically decreasing drug-associated side-effects. Therapeutic effects of developed drug targeting constructs will have to be tested in animal models of inflammation, in which E-selectin is expressed during the course of the disease. In this study several murine models of inflammation were investigated regarding expression of E-selectin.


METHODS
E-selectin expression was determined both at the mRNA level using RT-PCR and at the protein level by immunohistochemistry using two monoclonal antibodies (10E9.6 and MES-1). The models studied included delayed type hypersensitivity induced skin inflammation, dextran sodium sulphate induced colitis, kidney ischemia/reperfusion injury, atherosclerosis in ApoE knockout mice, and collagen induced arthritis.


RESULTS
In all animal models E-selectin mRNA expression was detected, although to a different extent. In contrast, only the delayed type hypersensitivity model and, to a minor extent, the collagen induced arthritis model showed E-selectin protein expression.


CONCLUSIONS
These results stress the need to determine E-selectin protein expression and not only mRNA expression, when choosing an animal model for testing E-selectin directed drug targeting preparations. In addition, in the arthritis model, E-selectin protein detection was dependent on the particular anti-E-selectin antibody used. This finding may not only have implications for the development and/or choice of homing devices to be used in E-selectin directed drug targeting preparations, but also for inflammation research in general.
Department of Pharmacokinetics & Drug Delivery, University Centre for Pharmacy, Groningen University Institute for Drug Exploration (GUIDE), The Netherlands.
M
M
Everts
S A
SA
Asgeirsdóttir
R J
RJ
Kok
J
J
Twisk
B
B
de Vries
E
E
Lubberts
E J
EJ
Bos
N
N
Werner
D K F
DK
Mejer
G
G
Molema
D000818:Animals; D004195:Disease Models, Animal; D019040:E-Selectin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007150:Immunohistochemistry; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000378:metabolism / Q000473:pathology
D019040:E-Selectin; D012333:RNA, Messenger