Quantitative Analysis of Cardiac 3-L-nitrotyrosine During Acute Allograft Rejection in an Experimental Heart Transplantation

Background: Recent studies have shown that nitric oxide interacts with superoxide to form peroxynitrite, a potent oxidant that modifies cellular proteins producing 3-L-nitrotyrosine (N-Tyr). This study was designed to evaluate N-Tyr quantitatively with high-performance liquid chromatography (HPLC) during cardiac allograft rejection.

Methods: Rat transplanted hearts (allogeneic or syngeneic grafts) were examined with HPLC analysis, immunohistochemistry for N-Tyr, and histological studies on 0, 1, 3, and 7 days after transplantation.

Results: No histological rejection was found in syngeneic grafts, or day 0 or 1 allografts. HPLC demonstrated that N-Tyr in allografts increased on day 1 and continued to increase through day 7, while N-Tyr was not detected in any syngeneic grafts. Immunostaining of the allografts did not show N-Tyr on day 1.

Conclusion: These results demonstrate that N-Tyr shows a time-dependent accumulation in cardiac allografts during acute rejection. N-Tyr detection using HPLC may be an useful maker for early diagnosis of acute rejection before pathological rejection occurs.