The goal of this project is to define the cellular and molecular mechanisms that contribute to lung injury after marrow transplantation. Our overall hypothesis is that cellular and molecular events involved in the pathogenesis of lung injury are determined by elements of the transplant procedure that activate inflammatory and immune responses and promote pneumonitis. Even in the absence of overt pneumonitis, these mechanisms may prime the lung for an exaggerated response to subsequent lung injury. Our experimental approach primarily will involve investigation of a murine model of marrow transplantation with graft versus host disease (GVHD). The model provides an opportunity to examine the cellular and molecular evolution of lung injury while controlling components of the transplant procedure. Studies in patients with idiopathic (non-infectious) pneumonia will be undertaken to investigate the role of similar mechanisms in the clinical setting. We have elected to focus on the specific hypothesis that GVHD activates proinflammatory cytokine production that results in priming of alveolar macrophages. Subsequent challenge with lipopolysaccaride (LPS) results in exaggerated release of tumor necrosis factor (TNF)-alpha and (either directly or indirectly) increased production of mitogenic and fibrogenic cytokines (i.e transforming growth factor (TGF)-alpha and TGF-beta. These latter events may be potentiated by total body irradiation. Our plans to investigate this hypothesis are outlined in the following specific aims.
Aim A. Determine the components of marrow transplant procedure that promote lung injury in a murine model of marrow transplantation. The incidence and severity of pneumonitis will be compared among mice in which irradiation dose and donor T-cell dose are varied.
Aim B. Analyze the cellular and molecular evolution of pneumonitis in the murine model. The following parameters will be examined in specimens obtained at intervals after marrow transplantation: l. Interferon levels in lavage and serum; 2. LPS and lipopolysaccharide binding protein (LBP) in serum and lavage and CD14 receptor expression in lavage and alveolar macrophages; 3. TNF-alpha immunoreactivity in lavage and serum and steady state mRNA levels in lung tissue; and spontaneous and LPS-triggered release of TNF by cultured alveolar macrophages obtained at intervals after marrow transplantation; 4. TGF-alpha, TGF-beta, procollagen steady state mRNA levels and immunolocalization in lung tissue.
Aim C. Determine whether susceptibility to LPS-induced lung injury is modified by marrow transplantation. Animals will be transplanted with a radiation regimen and T-cell dose lower than that associated with pneumonitis (determined in Aim A). They will then be challenged with LPS and parameters of lung injury and cytokine expression will be analyzed.
Aim D. Analysis of cytokine levels in patients with idiopathic pneumonia. Understanding the components of the transplant procedure that contribute to pneumonitis as well as cellular and molecular mechanisms evoked by these events could constitute a scientific foundation for fixture modification of transplant procedures, interventions aimed at minimizing the risk for lung injury, and direct therapeutic measures for idiopathic pneumonia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL055200-01
Application #
2233745
Study Section
Special Emphasis Panel (ZHL1-CSR-M (M2))
Project Start
1995-09-15
Project End
1999-08-31
Budget Start
1995-09-15
Budget End
1996-08-31
Support Year
1
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109
Clark, Joan G; Mandac-Dy, Janis B; Dixon, Anne E et al. (2004) Trafficking of Th1 cells to lung: a role for selectins and a P-selectin glycoprotein-1-independent ligand. Am J Respir Cell Mol Biol 30:220-7
Lorenz, E; Schwartz, D A; Martin, P J et al. (2001) Association of TLR4 mutations and the risk for acute GVHD after HLA-matched-sibling hematopoietic stem cell transplantation. Biol Blood Marrow Transplant 7:384-7
Dixon, A E; Mandac, J B; Madtes, D K et al. (2000) Chemokine expression in Th1 cell-induced lung injury: prominence of IFN-gamma-inducible chemokines. Am J Physiol Lung Cell Mol Physiol 279:L592-9
Dixon, A E; Mandac, J B; Martin, P J et al. (2000) Adherence of adoptively transferred alloreactive Th1 cells in lung: partial dependence on LFA-1 and ICAM-1. Am J Physiol Lung Cell Mol Physiol 279:L583-91
Clark, J G; Mandac, J B; Dixon, A E et al. (2000) Neutralization of tumor necrosis factor-alpha action delays but does not prevent lung injury induced by alloreactive T helper 1 cells. Transplantation 70:39-43
Clark, J G; Madtes, D K; Martin, T R et al. (1999) Idiopathic pneumonia after bone marrow transplantation: cytokine activation and lipopolysaccharide amplification in the bronchoalveolar compartment. Crit Care Med 27:1800-6
Clark, J G; Madtes, D K; Hackman, R C et al. (1998) Lung injury induced by alloreactive Th1 cells is characterized by host-derived mononuclear cell inflammation and activation of alveolar macrophages. J Immunol 161:1913-20