The Effect of A 2A Adenosine Receptor Agonist on Composite Tissue Allotransplant Survival: An In Vivo Preliminary Study

Abstract

In vitro experimental studies showed that A2a receptor agonists reduce allostimulatory functions of the dendritic cells through modulation of surface expression of the costimulatory molecules and down-regulation of cytokines. Similar suppressive effects on dendritic cells have also been observed with Cyclosporine A (CsA). In this study we sought to explore if combined use of these two drugs in vivo could prolong the survival of composite tissue allografts across MHC barriers. A total of 24 hindlimb transplantations were performed across a major MHC barrier from Brown Norway rats (BN; RT1n) to Lewis rats (Lew; RT1l) In the control group, either isografts (group 1, n = 2) or allografts (group 2, n = 3) were performed and no treatment was given. In the experimental group three kinds of treatment protocols were used: (1) A2a adenosine receptor agonist alone (group 3, n = 6); (2) CsA alone (group 4, n = 7); and (3) A2a adenosine receptor agonist + CsA combined treatment (group 5, n = 6). Mean survival times of each group as well as cytokine levels including IL-2, IL-4, IL-10, INF-gamma, and TNF-alpha were analyzed by ELISA. Isografts survived indefinitely. The mean survival times for allograft groups (group 2 to 5) were 9.8 +/- 1.3, 10.5 +/- 1.0, 29.8 +/- 1.7, and 22 +/- 1.4 days, respectively. Statistically, there was no difference between the allograft control group and the A2a adenosine receptor agonist treated group (P = 0.35). However, survival of the allografts in the CsA-treated group was significantly higher than the A2a adenosine receptor agonist treated (P < 00001) and combined CsA + A2a adenosine receptor agonist treated group (P < 0.0001). In vivo A2a adenosine receptor agonist treatment alone increased the levels of IL-2, INF-gamma, and TNF-alpha, which are important cytokines for induction of allotransplant rejection. However A2a adenosine receptor agonist in combination with CsA significantly reduced the levels of suppressor cytokines IL-4 and IL-10. As opposed to the previous in vitro studies, the results from this in vivo study showed that A2a adenosine receptor agonist treatment does not prolong composite tissue allograft survival. Compared to CsA treatment alone, the allotransplant survival was even shorter with the combined treatment. Treatment with A2a adenosine receptor agonist possibly promotes the differentiation of alloreactive CD4 cells predominantly into T-helper 1 phenotype. As a result, the levels of stimulatory cytokines contributing to allograft rejection are increased and suppressor cytokines are reduced, leading to accelerated allograft rejection.