Interactions between leukocytes, platelets and the endothelium in vein graft failure

Autologous venous grafts are preferred conduits in bypass surgery due to their length, easy harvesting, and feasibility. However, development of intimal hyperplasia decreases long-term patency rate in venous grafts prompting close clinical surveillance and further intervention. Mechanical forces, inflammation, and shear stress responses in venous grafts after surgical transfer may be involved in the development of intimal hyperplasia. This thesis focuses on inflammatory reactions in venous grafts. We hypothesized that interactions between leukocytes, platelets and endothelial cells induce intimal hyperplasia resulting in venous graft failure.

We demonstrate that VGs obtained by end-to-end anastomosis of the inferior vena cava (IVC) from donor mice grafted to the abdominal aorta in recipients suffer extensive endothelial injury, platelet deposition and leukocyte invasion early after grafting. Re-endothelialization of interposed venous grafts was completed after 4 weeks. Regenerated endothelium displayed normal resistance to recruitment of leukocytes. Thus, we found that there is a time window during the first weeks following grafting in which venous grafts are susceptible to vessel injury and inflammation.

Leukocyte recruitment on inflammatory endothelium initiates with margination, capture followed by rolling, firm adhesion and transmigration. We hypothesized that absence or functionblockage of leukocyte adhesion molecules P-selectin and E-selectin that are important for rolling may influence development of intimal hyperplasia in venous grafts by reduction of the recruitment of leukocytes. Indeed, intimal hyperplasia was significantly reduced in E- and P-selectin double deficient mice compared to WT both at 28 days and 63 days after VG transfer. This was paralleled by a reduction in the recruitment of leukocytes to the graft wall. Similar findings were made in WT mice treated with a combination of function-blocking antibodies against P- and E-selectin. The effect of P-selectin alone was addressed using P-selectin deficient mice. We found that intimal hyperplasia was significantly attenuated in mice deficient in P-selectin compared to WT mice 28 days after surgery concomitant with decreased leukocyte invasion. Interestingly, single-dose blockage of P-selectin or its leukocytic ligand PSGL-1 at the time of surgery could block the function of these molecules for up to 10 days and reduced later formation of intimal hyperplasia. Hence, early inhibition of these molecules has potential therapeutic effects on long-term vein graft failure. Platelets strongly influence haemostasis, inflammation and tissue regeneration after vascular injury, which are all represented in the period of re-endothelialization after venous graft transfer. We tested whether absence or function-blockage of platelets could influence intimal hyperplasia in venous grafts. We first used antiserum depletion of platelets for 12 days following surgical grafting. Indeed, IH and leukocyte invasion were reduced in platelet-depleted mice. Moreover, inhibition of integrin αIIbβ3, the main aggregation receptor on platelets, also reduced IH and leukocyte recruitment in the graft wall. The reduction of intimal hyperplasia in mice treated with the antibody against integrin αIIbβ3 was paralleled by radical reductions of the number of adherent platelets and leukocytes on the luminal surface of grafts one hour following grafting.

In summary, our work emphasizes the role of interactions between leukocytes, platelets and the vessel wall in venous grafts during their adaptation to the arterial circulation. These mechanisms constitute attractive targets for the development of further preventive pharmacological strategies against vein graft failure.

List of scientific papers

I. Tseng CN, Karlöf E, Chang YT, Lengquist M, Rotzius P, Berggren PO, Hedin U, Eriksson EE. Contribution of endothelial injury and inflammation in early phase to vein graft failure: The causal factors impact on the development of intimal hyperplasia in murine models. PLoS One. 2014;9:e98904.
https://doi.org/10.1371/journal.pone.0098904

II. Tseng CN, Chang YT, Lengquist M, Kronqvist M, Hedin U, Eriksson EE. Single-dose inhibition of P-selectin/P-selectin glycoprotein ligand-1 reduces intimal hyperplasia in mouse vein grafts. [Submitted]

III. Tseng CN, Chang YT, Lengquist M, Kronqvist M, Hedin U, Eriksson EE. Platelet adhesion on endothelium early after vein grafting mediates leukocyte recruitment and intimal hyperplasia in a murine model. Thromb Haemost. 2014;113(3).
https://doi.org/10.1160/TH14-07-0608