EXPRESSION OF CD40 AND GRANZYME B IN BRONCHOALVEOLAR LAVAGE CELLS FROM HUMAN LUNG TRANSPLANT RECIPIENTS

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TITLE: EXPRESSION OF CD40 AND GRANZYME B IN BRONCHOALVEOLAR LAVAGE CELLS FROM HUMAN LUNG TRANSPLANT RECIPIENTS

Ruili Shi,¹ Nancy Steward,¹ Andres Jaramillo,¹ Aviva Aloush,² Junbao Yang,¹ Elbert Trulock,² G. Alexander Patterson,³ T. Mohanakumar.¹

¹Surgery, Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA; ²Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA; ³Division of Cardiothoracic Surgery, Washington University School of Medicine, St. Louis, MO, USA

The goal of this study was to investigate the expression of CD40 and granzyme B in mononuclear cells from bronchoalveolar lavage (BAL) fluid during lung allograft rejection. The expression of CD40 on alveolar macrophages was detected at mRNA level by RT PCR and protein level by FACS analysis. The expression of granzyme B mRNA on total BAL mononuclear cells was performed using competitive–quantitative RT PCR. The data of FACS analysis and RT PCR were analyzed by Fisher’s Exact test and t–test, respectively. Forty–three BAL samples (28 without rejection and 15 with acute rejection) were analyzed for the expression of CD40 mRNA level. The relative expression levels of CD40 mRNA for the non–rejection group was 1.0 ± 0.16 and 1.17 ± 0.33 for the acute rejection group (P = 0.033, t–test). Eighteen patients were analyzed for CD40 expression by FACS; 11 without rejection and 7 with acute rejection. No patients in non–rejection group had detectable CD40–positive cells (P = 0.011, Fisher’s Exact test). In contrast, 4 patients in the acute rejection group had detectable levels of CD40–positive cells. The mean levels of CD40–positive cell in the rejection group was 14.90% ± 15.12. The mRNA levels of granzyme B were analyzed in 9 patients with non–rejection and 9 patients with acute rejection. The relative concentrations of mRNA were 0.14 pg/ul ± 0.12 for non–rejection and 1.6 pg/ul ± 2.08 for rejection groups (P = 0.046, t–test). These results demonstrate that increased expression of CD40 and granzyme B in BAL cells correlate with acute rejection following lung transplantation.