ABSTRACT
Administration of plasmid/lipid complexes to the lung airways may be associated, in addition to expression of transgene, with a range of other responses. We report here the induction of cytokines and cellular influx in the lung airway following intratracheal administration of an N-[1-(2-3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride/cholesterol/plasmid positively charged complex in mice. We show that 1) the appearance of the Th1-associated cytokines IFN-gamma and IL-12 in bronchoalveolar lavage fluid is caused by unmethylated CpG dinucleotide sequences present within the plasmid, and is enhanced by the lipid formulation; 2) cationic lipids by themselves do not induce IL-12 or IL-12p40; 3) TNF-alpha is rapidly induced by cationic lipids and plasmid/lipid complex, but not by plasmid alone; 4) an acute cellular influx is induced by cationic lipid alone and by a plasmid/lipid complex, but to a much lesser extent by plasmid alone; and 5) plasmid methylation does not influence the degree of inflammatory cell influx. The induction of the innate immune responses by plasmid/lipid complexes may be advantageous to gene therapy of lung diseases. In particular, induction of the Th1 cell-promoting cytokines by plasmid/lipid complexes could, in conjunction with an expressed transgene, be used to modulate immune responses in the lung airways in disease conditions that are deficient in Th1 cell responses or that have a dominant Th2 phenotype. Alternatively, the elimination of immunostimulatory sequences in plasmids may improve the tolerability and/or efficacy of nonviral gene therapy, especially for diseases requiring chronic administration.
Subject(s)
Cholesterol/immunology , Cytokines/immunology , Genetic Vectors/adverse effects , Lung/immunology , Lymphocyte Activation , Plasmids/immunology , Th1 Cells/immunology , Th2 Cells/immunology , Administration, Inhalation , Animals , Cholesterol/administration & dosage , Cholesterol/analogs & derivatives , Gene Transfer Techniques/adverse effects , Humans , Male , Mice , Mice, Inbred C57BL , Plasmids/administration & dosage , Plasmids/geneticsABSTRACT
The occurrence of a congenital cleft palate greatly impedes the habituation of efficient deglutition of an infant, causing aspiration of fluids in the air passage which may lead to complications like bronchopneumonia and infection of the airway and lungs. Inadequate nourishment due to difficulty in feeding affects the health and acts as a stumbling block in the milestones of normal development. Severe nutritional deficiencies further complicate surgical closure at the right time. A palatal obturator given to the infant effectively separates the oral cavity from the nasal cavity and is of great help in feeding till the defect can be surgically repaired.
Subject(s)
Palatal Obturators , Bronchopneumonia/etiology , Cleft Palate/complications , Cleft Palate/therapy , Dental Prosthesis Design , Humans , Infant , Infant Nutrition Disorders/etiology , MaleABSTRACT
A simple colorimetric method is described for the determination of metaproterenol sulfate (orciprenaline sulfate). The method is based on measurement of a colored species formed when metaproterenol sulfate is treated with diazotized dapsone, p-nitroaniline, or benzocaine at room temperature, followed by treatment with an aqueous solution of trimethylamine in the case of benzocaine. Compounds such as starch, talc, and common excipients do not interfere in the reaction. Statistical validation showed that the method was precise and accurate. The results agree well with those obtained by other methods reported in the literature.
