title: Increased levels of T cell granzyme b in bronchiolitis obliterans syndrome are not suppressed adequately by current immunosuppressive regimens.
abstract: Bronchiolitis obliterans syndrome (BOS) is characterized by persistent alloreactive, infective and non-specific epithelial injury, loss of epithelial integrity and dysregulated repair. We have reported increased apoptosis of epithelial cells collected from the large airway in lung transplant recipients. As part of the alloreactive response, T cells induce apoptosis of target epithelial cells by secreting granzyme b. We hypothesized that granzyme b would be increased in lung transplant patients with acute rejection and BOS and that commonly used immunosuppressive agents would fail to suppress this serine protease adequately. We investigated intracellular T cell granzyme b in blood, bronchoalveolar lavage (BAL) and large airway brushing (23 controls, 29 stable transplant, 23 BOS, 28 acute rejection, 31 infection) using flow cytometry and assessed the effect of clinically relevant concentrations of cyclosporin A, tacrolimus, methylprednisolone and a protease inhibitor, gabexate mesilate, on in vitro granzyme b production. Granzyme b was increased significantly in all compartments of all transplant groups compared to controls. Surprisingly, granzyme b was even higher in patients with BOS than in patients with acute rejection. In longitudinal analysis in three patients, blood granzyme b increased prior to or at the onset of BOS. In vitro, methylprednisolone and gabexate mesilate had no effect and cyclosporin A and tacrolimus only a moderate effect on production of granzyme b by CD8(+) T cells. Increased T cell granzyme b production may contribute to BOS pathogenesis and is not curtailed by current immunosuppressants. Longitudinal investigation of granzyme b in blood may provide an adjunctive non-invasive method for predicting BOS/OB.
authors.affiliation: Lung Research Laboratory, Hanson Institute, Adelaide, SA, Australia. sandy.hodge@imvs.sa.gov.au
authors.forename: S
authors.initials: S
authors.lastname: Hodge
authors.forename: G
authors.initials: G
authors.lastname: Hodge
authors.forename: J
authors.initials: J
authors.lastname: Ahern
authors.forename: C-L
authors.initials: CL
authors.lastname: Liew
authors.forename: P
authors.initials: P
authors.lastname: Hopkins
authors.forename: D C
authors.initials: DC
authors.lastname: Chambers
authors.forename: P N
authors.initials: PN
authors.lastname: Reynolds
authors.forename: M
authors.initials: M
authors.lastname: Holmes
mesh_terms: D000328:Adult; D000368:Aged; D001989:Bronchiolitis Obliterans / Q000188:drug therapy / Q000201:enzymology* / Q000209:etiology / Q000276:immunology; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D001999:Bronchoscopy / Q000379:methods; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006084:Graft Rejection / Q000201:enzymology / Q000276:immunology; D053804:Granzymes / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology / Q000627:therapeutic use*; D008137:Longitudinal Studies; D016040:Lung Transplantation / Q000009:adverse effects / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000201:enzymology*
chemical_list: D007166:Immunosuppressive Agents; D053804:Granzymes