THE IMPACT OF AEROSOL CYCLOSPORINE ON PROINFLAMMATORY CYTOKINE GENE EXPRESSION IN LUNG TRANSPLANT RECIPIENTS.
K Spichty, M Pavalakis, J Cai, A Iacono, J Dauber, W Grgurich, R Keenan, G Burckart, K McCurry, B Griffith and A Zeevi University of Pittsburgh, Pittsburgh, PA.
A randomized study was initiated to determine whether aerosolized CsA given as prophylaxis can affect the frequency of acute rejection. We analyzed the effect of ACsA on intragraft cytokine gene expression for Granzyme B (a marker for cytotoxic T cells) IL-15 (a T cell growth factor) RANTES (a chemokine for monocytes and activated T cells) and FasL in 12 lung transplant recipients with ACsA, 16 controls (no ACsA) and in 10 patients with CMV pneumonia. Quantitative RT-PCR for intragraft cytokine expression was performed on bronchoalveolar lavage cells (BAL) from ACsA (48 BALs) and control (45 BALs) patients during various clinical conditions. Patient derived cDNA (WT) was co-amplified with a known amount of target gene competitor (CT) which is shorter and is amplified by the same primers. The results for each gene were standardized for the housekeeping gene GAPDH. A ratio of GB/GAPDH less than 40 x 10-3 was significantly (p < 0.0001) associated with quiescence in the ACsA group. Only 5% (2/38 BAL samples) of patients in the ACsA group exhibited GB/GAPDH greater than 40 x 10-3 during clinical quiescence. In contrast, 17
% of BAL samples in the control group exhibited GB/GAP ratio greater than 40 x 10-3.
These results indicate that ACsA may better control the intragraft activation events than systemic immunosuppression alone. CMV infection may also induce the up-regulation of GB. In 7 patients with refractory rejection and CMV disease, the GB/GAP was significantly higher (mean 400 ± 407 x 10-3 vs. 12 ± 16 x 10-3) than in patients with CMV disease only. None of the lung transplant recipients with CMV disease exhibited FasL but it was readily detectible in lung transplant recipients with allograft rejection. IL-15 and RANTES mRNA were up-regulated during rejection in both the ACsA and control groups. In summary, the presence of ACsA during clinical quiescence and post rejection treatment seems to better control the intragraft cytokine activation.