Immunologic reactivity to lipid-DNA conjugates has traditionally been viewed as less of an issue than with viral vectors. We performed a dose escalation safety trial of aerosolized cystic fibrosis transmembrane conductance regulator (CFTR) cDNA to the lower airways of eight adult cystic fibrosis patients, and monitored expression by RT-PCR. The cDNA was complexed to a cationic lipid amphiphile (GL-67) consisting of a cholesterol anchor linked to a spermine head group. CFTR transgene was detected in three patients at 2-7 days after gene administration. Four of the eight patients developed a pronounced clinical syndrome of fever (maximum of 103.3EF), myalgias, and arthralgia beginning within 6 hr of gene administration. Serum IL-6 but not levels of IL-8, IL-1, TNF-alpha, or IFN-gamma became elevated within 1-3 hr of gene administration. No antibodies to the cationic liposome or plasmid DNA were detected. We found that plasmid DNA by itself elicited minimal proliferation of peripheral blood mononuclear cells taken from study patients, but led to brisk immune cell proliferation when complexed to a cationic lipid. Lipid and DNA were synergistic in causing this response. Cellular proliferation was also seen with eukaryotic DNA, suggesting that at least part of the immunologic response to lipid-DNA conjugates is independent of unmethylated (E. coli-derived) CpG sequences that have previously been associated with innate inflammatory changes in the lung.

A reversible electronic switching effect has been observed in plasma polymer films with embedded silver nanoparticles. The electrical and nanostructural properties of the films have been investigated, and three different structure types were observed: metallic, percolation, and dielectric. While for the metallic and dielectric types, respectively, metallic conduction and thermally activated tunneling can be identified as the dominant electronic conduction mechanisms, switching appears only in percolation structures. These drastic, abrupt changes of up to six orders of magnitude in the current-voltage behavior are highly reversible for these nanocomposite materials, and are defined as threshold switching

Cationic lipids show promise as vectors for transfer of CFTR cDNA to airway epithelia of patients with cystic fibrosis (CF). However, previous studies have not compared the effect of DNA-lipid to DNA alone. Recently, we developed a formulation of plasmid encoding CFTR (pCF1-CFTR) and cationic lipid (GL-67:DOPE) that generated greater gene transfer in mouse lung than previously described DNA-lipid vectors. Therefore, we tested the hypothesis that DNA-lipid complexes were more effective than DNA alone at transferring CFTR cDNA to airway epithelia in vivo. We administered complexes of DNA-lipid to one nostril and DNA alone to the other nostril in a randomized, double-blind study. Electrophysiologic measurements showed that DNA-lipid complexes partially corrected the Cl

Cystic fibrosis (CF) is caused by a mutation of the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). We examined platelet function in CF patients because lung inflammation is part of this disease and platelets contribute to inflammation. CF patients had increased circulating leukocyte-platelet aggregates and increased platelet responsiveness to agonists compared with healthy controls. CF plasma caused activation of normal and CF platelets; however, activation was greater in CF platelets. Furthermore, washed CF platelets also showed increased reactivity to agonists. CF platelet hyperreactivity was incompletely inhibited by prostaglandin E 1 (PGE 1 ). As demonstrated by Western blotting and reverse-transcriptase-polymerase chain reaction (RT-PCR), there was neither CFTR nor CFTR-specific mRNA in normal platelets. There were abnormalities in the fatty acid composition of membrane fractions of CF platelets. In summary, CF patients have an increase in circulating activated platelets and platelet reactivity, as determined by monocyte-platelet aggregation, neutrophilplatelet aggregation, and platelet surface P-selectin. This increased platelet activation in CF is the result of both a plasma factor(s) and an intrinsic platelet mechanism via cyclic adenosine monophosphate ( IntroductionCystic fibrosis (CF) is the most common lethal genetic disease in whites, affecting approximately 1 in 2500 live births. 1 In 1989, Kerem et al 2 ; Riordan et al 3 ; and Rommens et al 4 identified the genetic mutation responsible for CF and cloned its protein product, which is called the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is a member of the adenosine triphosphate (ATP)-binding cassette family of transporters and acts chiefly as a chloride channel, although it has various other functions including regulation of alternate chloride channels and epithelial sodium channels, and transport of ATP. [5][6][7] Defects in CFTR cause reduced chloride secretion into airways and enhanced sodium reabsorption, thereby resulting in dehydrated airway secretions, poor mucociliary clearance, and airway obstruction. 1 Obstruction of the airways, along with poorly understood altered host defense mechanisms and an enhanced inflammatory response, leads to chronic airway infection and inflammation. This, in turn, causes an influx of polymorphonuclear cells that, when they become senescent and release their DNA into the airway mucous, contribute to thickening of airway secretions. 8 The self-perpetuating cycle of obstruction, infection, and inflammation eventually causes pulmonary parenchymal destruction and respiratory failure. The life expectancy for CF patients in the United States is currently approximately 33 years. 9 Previous studies have shown that CF patients have increased ex vivo platelet aggregability, increased release of thromboxane A 2 (TXA 2 ), 10-12 and a blunted response to prostaglandin E 1 (PGE 1 )-induced inhibition of platelet aggregation. 13,14 Platelets contain or generate many inf...