Construction of single-chain interleukin-12 DNA plasmid to treat airway hyperresponsiveness in an animal model of asthma
Journal
Human Gene Therapy
Journal Volume
12
Journal Issue
17
Pages
2065-2079
Date Issued
2001
Author(s)
Abstract
Allergic asthma is strongly associated with the airway inflammation caused by the dysregulated production of cytokines secreted by the allergen-specific type-2 T helper (Th2) cells. Interleukin (IL)-12 is a heterodimeric cytokine, which strongly promotes the differentiation of naive CD4+ T cells to the type-1 T helper (Th1) phenotype and suppresses the expression of Th2 cytokines. Therefore, immunotherapy with IL-12 has been suggested as a possible therapy for asthma. In previous studies, we developed a murine model of airway inflammation based on the purified, house dust-mite allergen Der p 1 (Dermatophagodies pteronyssinus) as a clinically relevant allergen. We hypothesized that the expression of IL-12 in the airway may represent an effective therapy for allergic airway diseases. In this study, we investigate whether the local transfer of the IL-12 gene to respiratory tissues modifies allergic inflammation and airway hyper-responsiveness (AHR) in our disease model. To enhance the in vivo delivery of the IL-12 gene, we expressed the murine single-chain IL-12 protein from a nonviral vector to which the two IL-12 subunits (p35 and p40) were linked by a 14- to 18-amino-acid linker. One of these single-chain IL-12s, containing an 18 amino-acid polypeptide linker, was stably expressed and had a high level of biological activity comparable to that of native IL-12 in vitro. In mice with Der p 1-induced asthma, the local administration of this IL-12 fusion gene into the lungs significantly prevented the development of AHR, abrogated airway eosinophilia, and inhibited type-2 cytokine production. These findings indicate that the local transfer of the single-chain IL-12 gene is effective in modulating pulmonary allergic responses and may be a convenient method for future applications of DNA vaccination.
SDGs
Other Subjects
amino acid; gamma interferon; house dust allergen; interleukin 12; interleukin 5; protein p35; protein p40; virus vector; airway; animal cell; animal model; animal tissue; article; asthma; cell differentiation; cell infiltration; controlled study; cytokine production; drug activity; drug effect; drug efficacy; eosinophilia; female; fusion gene; gene targeting; gene transfer; immunotherapy; lung parenchyma; mouse; nonhuman; phenotype; plasmid; protein expression; respiratory tract disease; T lymphocyte; Th1 cell; Allergens; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; COS Cells; Disease Models, Animal; Female; Gene Therapy; Immunotherapy; Inflammation; Interleukin-12; Lung; Methacholine Chloride; Mice; Mice, Inbred C57BL; Plasmids; Transfection; Vaccines, DNA; Acari; Animalia; Astigmata; Murinae; Pyroglyphidae
Type
journal article