Bacterial pneumonia is a frequent cause of morbidity and mortality in persons abusing alcohol. One proposed mechanism of the susceptibility of the host to pneumonia is a diminished release of cytokines, specifically Tumor Necrosis Factor in the lung of alcoholics in response to a bacterial challenge. Our previous work has demonstrated that TNF is suppressed in vivo in acutely alcoholic animals and that increasing TNF in vivo by pre- treatment of alcoholic animals with interferon-gamma augments pulmonary host defense. The goals of this project are two-fold: 1) to investigate the molecular mechanisms of ethanol's effect on TNF gene expression and 2) to utilize gene therapy to modulate TNF responses in the alcoholic host to augment the immune response to intrapulmonary pathogens. It is our hypothesis that acute ethanol downregulates TNF through a posttrancriptional mechanism with a resultant impairment of pulmonary host defense. Moreover, we hypothesize that overcoming this post- transcriptional block by gene therapy will improve pulmonary host defense to intrapulmonary pathogens. This proposal will test this hypothesis using a rat model and has three specific aims: 1) to characterize the effect of ethanol on TNF gene regulation and protein processing in vivo, 2) To assess the ability of recombinant adenoviral vectors encoding TNF or Interferon-gamma to upregulate TNF in vitro and in vivo, and 3) to utilize gene therapy to upregulate intrapulmonary TNF in vivo to augment pulmonary host defense in alcohol suppressed animals challenged with intrapulmonary pathogens. To accomplish these aims we will: a) investigate the in vivo effect of ethanol on TNF mRNA levels and protein processing of this cytokine, b) to generate recombinant adenoviruses encoding human TNF and murine interferon-gamma, c) study the effect of ethanol on the expression of these two vectors in vitro and in vivo, d) investigate whether these vectors can augment recruitment of polymorphonuclear lymphocytes into the lung of alcoholic animals challenged with endotoxin and e) whether these vectors can augment bacterial killing in alcoholic animals. The identification of the precise block of TNF production will yield new insights into ethanol's effect on gene expression and the use of cytokine gene therapy will further elucidate the role of these cytokines in the alcoholic host as well as lead to potential new treatment strategies.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29AA010384-05
Application #
2894080
Study Section
Biochemistry, Physiology and Medicine Subcommittee (ALCB)
Program Officer
Isaki, Leslie
Project Start
1995-08-01
Project End
2001-05-31
Budget Start
1999-08-01
Budget End
2001-05-31
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Louisiana State University Hsc New Orleans
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
782627814
City
New Orleans
State
LA
Country
United States
Zip Code
70112
Wang, J; Cheung, A T; Kolls, J K et al. (2001) Effects of adenovirus-mediated liver-selective overexpression of protein tyrosine phosphatase-1b on insulin sensitivity in vivo. Diabetes Obes Metab 3:367-80
Forlow, S B; Schurr, J R; Kolls, J K et al. (2001) Increased granulopoiesis through interleukin-17 and granulocyte colony-stimulating factor in leukocyte adhesion molecule-deficient mice. Blood 98:3309-14
Cheung, A T; Wang, J; Ree, D et al. (2000) Tumor necrosis factor-alpha induces hepatic insulin resistance in obese Zucker (fa/fa) rats via interaction of leukocyte antigen-related tyrosine phosphatase with focal adhesion kinase. Diabetes 49:810-9
Zhang, Z; Kolls, J K; Oliver, P et al. (2000) Activation of tumor necrosis factor-alpha-converting enzyme-mediated ectodomain shedding by nitric oxide. J Biol Chem 275:15839-44
Cheung, A T; Ree, D; Kolls, J K et al. (1998) An in vivo model for elucidation of the mechanism of tumor necrosis factor-alpha (TNF-alpha)-induced insulin resistance: evidence for differential regulation of insulin signaling by TNF-alpha. Endocrinology 139:4928-35
Kolls, J K; Lei, D; Stoltz, D et al. (1998) Adenoviral-mediated interferon-gamma gene therapy augments pulmonary host defense of ethanol-treated rats. Alcohol Clin Exp Res 22:157-62
Schwarzenberger, P; Lei, D; Freeman, S M et al. (1998) Antitumor activity with the HSV-tk-gene-modified cell line PA-1-STK in malignant mesothelioma. Am J Respir Cell Mol Biol 19:333-7
Cheng, D Y; Kolls, J K; Lei, D et al. (1997) In vivo and in vitro gene transfer and expression in rat intestinal epithelial cells by E1-deleted adenoviral vector. Hum Gene Ther 8:755-64
Kolls, J K; Lei, D; Vazquez, C et al. (1997) Exacerbation of murine Pneumocystis carinii infection by adenoviral-mediated gene transfer of a TNF inhibitor. Am J Respir Cell Mol Biol 16:112-8
Lei, D; Lancaster Jr, J R; Joshi, M S et al. (1997) Activation of alveolar macrophages and lung host defenses using transfer of the interferon-gamma gene. Am J Physiol 272:L852-9

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