E-selectin early overexpression induced by allogeneic activation in isolated mouse lung.
BACKGROUND
The interaction between host lymphocytes and graft endothelial cells plays an important role in graft rejection.


METHODS
Using our model of isolated ventilated lung from female mouse perfused with fresh blood from either isogeneic or allogeneic male mouse for 3 hours without noticeable ischemia, we have investigated the kinetics of the early events after endothelial cell triggering by E-selectin engagement.


RESULTS
Isogeneic perfusion induced nonspecific endothelial cell activation, which was characterized by up-regulation of E-selectin, intercellular adhesion molecule (ICAM)-1, and of the pro-inflammatory cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, and lymphotoxin-alpha (mRNAs by real-time polymerase chain reaction). Allogeneic perfusion was characterized after 3 hours by an additional loose adhesion of lymphocytes mediated by the E-selectin and related to the allogeneic activation of endothelial cells. These in turn expressed the I-A molecule (immunostaining). ICAM-1 and lymphocyte function-associated antigen (LFA)-3 mRNA levels were significantly increased in lung extracts after 2 hours, then vascular cell adhesion molecule (VCAM)-1 and TNF-alpha mRNAs after 3 hours without evidence of TNF-alpha production (enzyme-linked immunoadsorbent assay). The major participation of the E-selectin in early allogeneic activation by way of the protein kinase (PK)C pathway was confirmed by using a neutralizing anti-CD62E monoclonal antibody or the inhibitory PKC 19-31 fragment.


CONCLUSIONS
Altogether, our results demonstrate that E-selectin expression (1) is not a consequence of TNF-alpha triggering, (2) up-regulates its own expression and expression of I-A, VCAM-1, TNF-alpha, and lymphotoxin-alpha mRNAs, and (3) down-regulates expression of LFA-3 and ICAM-1 mRNAs. In conclusion, in our physiologic model, the E-selectin highly participates in the loose adhesion of allogeneic lymphocytes and in the early activation of endothelial cell and therefore in structural and functional lung alterations.
Joucher
Franck
F
CNRS UMR 8078, I.P.S.C., Université Paris XI, Centre Chirurgical Marie-Lannelongue, Le Plessis-Robinson, France.
Mazmanian
Guy-Michel
GM
German-Fattal
Michele
M
D000818:Animals; D017209:Apoptosis; D018968:CD58 Antigens / Q000235:genetics; D002448:Cell Adhesion; D004268:DNA-Binding Proteins / Q000235:genetics; D019040:E-Selectin / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology*; D005260:Female; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics; D008168:Lung / Q000378:metabolism*; D008214:Lymphocytes / Q000502:physiology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009336:Necrosis; D009687:Nuclear Proteins / Q000235:genetics; D011493:Protein Kinase C / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics
D018968:CD58 Antigens; D004268:DNA-Binding Proteins; D019040:E-Selectin; D007376:Interleukin-2; D009687:Nuclear Proteins; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D011493:Protein Kinase C