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Adenovirus (Adv)-mediated gene transfer requires efficient infection of target cells. The objective of this study was to establish whether alveolar macrophages (AM) and T cells (AT) from sarcoid patients were permissive to infection with Adv vectors and if this property could be used to investigate cytokine gene regulation. Sarcoid and normal bronchoalveolar lavage (BAL) specimens infected with Adv vectors expressing either beta-galactosidase or a green fluorescent protein were analyzed for transgene expression by fluorescence-activated cell sorter (FACS) and direct immunofluorescence, respectively. Expression of surface antigens previously associated with Adv infection, the coxsackie/adenovirus receptor (CAR), alpha v beta 3, and alpha v beta 5 integrins, was also assessed using FACS analysis. Sarcoid AM and AT were found to efficiently express Adv transgenes, unlike AM from normal volunteers, peripheral blood monocytes, and peripheral blood T cells. Cells permissive to Adv infection expressed the CAR and alpha v beta 5 integrin (also alpha v beta 3 integrin for AM). The data indicate that the upregulation of Adv receptors and the ability to infect sarcoid AM and AT are related to the inflammatory environment within the lung. Having demonstrated efficient Adv-mediated transgene delivery to sarcoid AM and AT, a construct encoding porcine I kappa B alpha was then used to investigate the requirement for nuclear factor (NF)-kappa B in the regulation of cytokine gene expression in pulmonary sarcoidosis. Overexpression of I kappa B alpha in sarcoid BAL specimens indicated that tumor necrosis factor-alpha and interleukin (IL)-6 production by AM and interferon (IFN)-gamma production by AT is NF-kappa B dependent, whereas IL-4 production by AT is NF-kappa B independent. This is the first occasion that the requirement for NF-kappa B in IFN-gamma gene expression within primary human T cells has been demonstrated. The results of this study have implications for the future investigation of molecular pathways in inflammatory lung disease.

Original publication

DOI

10.1165/ajrcmb.25.2.4327

Type

Journal article

Journal

American journal of respiratory cell and molecular biology

Publication Date

08/2001

Volume

25

Pages

141 - 149

Addresses

Kennedy Institute of Rheumatology, London, United Kingdom.

Keywords

T-Lymphocytes, Macrophages, Alveolar, Humans, Adenoviridae, Lung Diseases, Sarcoidosis, Tumor Necrosis Factor-alpha, DNA-Binding Proteins, NF-kappa B, Macrophage Colony-Stimulating Factor, Integrins, Receptors, Vitronectin, Receptors, Virus, Interleukin-4, Interleukin-6, Cytokines, Gene Transfer Techniques, Signal Transduction, Gene Expression Regulation, Down-Regulation, Genetic Vectors, I-kappa B Proteins, Interferon-gamma, Coxsackie and Adenovirus Receptor-Like Membrane Protein, In Vitro Techniques, NF-KappaB Inhibitor alpha