Publications
297 results found
Griesenbach U, Kren BT, Stern M, et al., 2001, Conversion of wild-type CFTR to the G551D mutation in primary rat hepatocytes using RNA/DNA oligonucleotides, THORAX, Vol: 56, Pages: 44-45, ISSN: 0040-6376
Fox E, Griesenbach U, Rogers DF, et al., 2001, Successful oligodeoxynucleotide transfer to the airway epithelium following intravenous delivery, THORAX, Vol: 56, Pages: 2-2, ISSN: 0040-6376
Griesenbach U, Cassady RL, Cain RJ, et al., 2001, The effect of NFkB decoy oligonucleotides in two murine models of pulmonary inflammation, THORAX, Vol: 56, Pages: 45-45, ISSN: 0040-6376
Griesenbach U, Alton EWFW, 2001, Recent progress in gene therapy for cystic fibrosis, CURRENT OPINION IN MOLECULAR THERAPEUTICS, Vol: 3, Pages: 385-389, ISSN: 1464-8431
- Author Web Link
- Cite
- Citations: 5
Scheid P, Kempster L, Griesenbach U, et al., 2001, Inflammation in cystic fibrosis airways: relationship to increased bacterial adherence, EUROPEAN RESPIRATORY JOURNAL, Vol: 17, Pages: 27-35, ISSN: 0903-1936
- Author Web Link
- Cite
- Citations: 53
Griesenbach U, Judd D, Ferrari S, et al., 2000, Sendai virus-mediated overexpression of interleukin 10 following pulmonary and intramuscular adminstration, THORAX, Vol: 55, Pages: A5-A5, ISSN: 0040-6376
Yonemitsu Y, Kitson C, Ferrari S, et al., 2000, Efficient gene transfer to airway epithelium using recombinant Sendai virus, NATURE BIOTECHNOLOGY, Vol: 18, Pages: 970-973, ISSN: 1087-0156
- Author Web Link
- Cite
- Citations: 182
Stern M, Ulrich K, Robinson C, et al., 2000, Pretreatment with cationic lipid-mediated transfer of the Na<SUP>+</SUP>K<SUP>+</SUP>-ATPase pump in a mouse model in vivo augments resolution of high permeability pulmonary oedema, GENE THERAPY, Vol: 7, Pages: 960-966, ISSN: 0969-7128
- Author Web Link
- Cite
- Citations: 43
Griesenbach U, Scheid P, Hillery E, et al., 2000, Anti-inflammatory gene therapy directed at the airway epithelium, GENE THERAPY, Vol: 7, Pages: 306-313, ISSN: 0969-7128
- Author Web Link
- Cite
- Citations: 55
Kempster L, Griesenbach U, Gruenert DC, et al., 1999, Activation of a p50-containing NFκB complex in CF models, Thorax, Vol: 54, ISSN: 0040-6376
Chronic airway inflammation contributes significantly to progressive lung damage in CF. At present it is uncertain whether the inflammation is simply secondary to infection, or more directly related to the CFTR defect, or a combination of both. We have previously repotted that total NFκB activity and IL-8 secretion are similar in Cos7 cells transiently overexpressing wildtype and mutant CFTR cDNA and that partial restoration of ΔF508 CFTR trafficking in a homozygous CF cell line did not reduce either NFκB activation or IL-8 secretion. Using 'supershift' assays with several anti-Rel antibodies (p50, p52, p65, c-Rel and Rel-B), we have now assessed the composition of the NFκB DNA/protein complexes generated in etectrophoretic mobility shift assays (EMSA). In our hands, three specific DNA/protein complexes are produced. Of these, complexes I and II are increased after TNF-α and Pseudomonas aeruginosa (PA) stimulation in wildtype cell lines and mice respectively, and are therefore relevent to inflammation. We now demonstrate the presence of p50 protein in complex II in CF and non-CF cell lines. However, the composition of the other DNA/protein complexes could not be unambiguously identified. The degree of activation of the p50-containing complex in CF and non-CF cell lines was compared. CF cell lines derived from tracheal epithelium (ΣCFTE29o-) and from sub-mucosal epithelium (CFSMEo-) exhibited 6-fold and 4-fold increases in p50 activation respectively, when compared to a mean value of non-CF cells obtained from human tracheal epithelium (9HTEo-), tracheo-bronchial epithelium (TBEo-) and human bronchial epithelium (16HBE14o-) (n=6 p<0.001). A CF cell line derived from nasal polyp tissue (CFNPE29o-) exhibited a 2-fold decrease in p50 activation (n=6 p<0.001). IL-8 secretion was only significantly increased in ΣCFTE29o- cells when compared to the non-CF cell lines (CFTE: 7897 ± 1497 pg/mg protein, WT: 797 ± 69
Judd DV, Griesenbach U, Wells DJ, et al., 1999, Intramuscular gene transfer of interleukin-10 cDNA: Towards treatment for lung inflammation., THORAX, Vol: 54, Pages: A3-A3, ISSN: 0040-6376
Griesenbach U, Scheid P, Hillery E, et al., 1999, Developing anti-inflammatory lung gene therapy, THORAX, Vol: 54, Pages: A58-A58, ISSN: 0040-6376
Griesenbach U, Geddes DM, Alton EWFW, 1999, The pathogenic consequences of a single mutated CFTR gene, THORAX, Vol: 54, Pages: S19-S23, ISSN: 0040-6376
- Author Web Link
- Cite
- Citations: 26
Alton EWFW, Griesenbach U, Geddes DM, 1999, Gene therapy for asthma: inspired research or unnecessary effort?, GENE THERAPY, Vol: 6, Pages: 155-156, ISSN: 0969-7128
- Author Web Link
- Cite
- Citations: 7
Kempster L, Griesenbach U, Browning JE, et al., 1998, Relationship between ΔF508 CFTR protein accumulation in the endoplasmic reticulum, NFκB activity and interleukin-8 secretion, Thorax, Vol: 53, ISSN: 0040-6376
The clinical phenotype of cystic fibrosis lung disease is strongly related to infection and inflammation. Several reports have suggested that inflammation might precede infection and relate to an endogenous upregulation of proinflammatory mediators. Nuclear factor kappa B (NFκB) is a transcription factor that regulates expression of inflammatory mediators. It has previously been postulated that protein overload of the endoplasmic reticulum (ER) causes activation of NFκB. Retention of ΔF508 protein in the ER of CF patients might increase NFκB activity and therefore increase expression of pro-inflammatory mediators such as interleukin-8 (IL-8). To investigate if CFTR retention in the ER is related to NFκB activation and IL-8 secretion we undertook a number of experiments in which NFκB activity was measured through electrophoretic mobility shift assays (EMSA) and IL-8 through ELISA assays. (1) CF and non CP cells (CFTE and 16HBE cells, respectively and Cos7 cells) were incubated with TMAO (10 mM) for 72 hrs and chloride efflux was measured by SPQ fluorescence. TMAO produced a significant (p<0.05) increase in chloride efflux from 0.01 mMsec-1 ± 0.01 (n=7) to 0.48 mMsec-1 ± 0.22 (n=11) in CFTE cells. Despite this we could not detect any significant change in IL-8 activation or in IL-8 secretion in CFTE cells (no TMAO: 8110 ± 2286 pg/mg protein, TMAO: 6963 ± 2286 pg/mg protein n=4) or any of the other cell lines used. (2) Transient overexpression of ΔF508 protein should lead to increased amounts of incompletely processed CFTR protein in the ER when compared to overexpression of wildtype protein. Cos7 cells were transfected with various plasmids carrying either wildtype (WT), ΔF508 or G551D CFTR cDNA or a β-galactosidase reporter gene. Successful transfection of the CFTR cDNAs was confirmed by RT-PCR or by CPRG assay for the β-galactosidase reporter gene. However, NFκB activity a
Judd DV, Kitson C, Griesenbach U, et al., 1998, Development of a new in vivo model for pulmonary gene transfer, Thorax, Vol: 53, ISSN: 0040-6376
The ferret airway shows many anatomical similarities to human tissues including the number of submucosal glands and the basal level of mucus production. We have assessed the feasibility of gene transfer in this animal. Female ferrets were nasally instilled with increasing doses (800{n=3}, 2400{n=5}, 4000{n=3} & 5600{n=4}micrograms) of pCF1-CAT complexed with GL-67 at a lipid:DNA ratio of 1:4 under ketamine/xylazine anaesthesia. Transgene expression was assessed 48 hours after instillation. A dose dependent increase in transgene expresssion within the lung was observed with maximal expression of 21×10-6 CAT units/μg protein for the 4000μg dose. At the highest dose expression was reduced approximately 20-fold. The four right lobes consistently showed greater expression, probably due to the right bronchus being more dependent in the ferret. With increasing dose, transgene expression was observed in a greater number of lobes within an animal, and at the 4000ug dose all lobes showed transgene expression. Expression was consistently lower in the trachea at all doses. A further three ferrets were instilled with 4000μg of a plasmid encoding non-secreted alkaline phosphatase (NSAP) complexed with GL-67 to assess histological expression. NSAP expressing cells were observed only in the parenchyma of the lung. No expression was observed in airway epithelium or trachea. In order to assess the relevance of the ferret model with respect to other gene transfer vector systems three animals were instilled with 1010 pfu of Ad2/CMVβgal-4. 48 hours following treatment transgene expression was assessed. No significant expression was observed either quantitatively or histologically. Similarly in an ex vivo ferret trachea model, treatment with Ad2/CMVβgal-4 failed to achieve transgene expression. We conclude that liposome mediated gene transfer is feasible in vivo in the ferret. This may prove useful in the assessment of biological barriers to pulmonary gene tr
Stern M, Ulrich K, Copeland J, et al., 1998, Cationic liposome-mediated gene transfer of the Na+-K+-ATPase pump to the lungs in vivo significantly decreases pulmonary oedema, Thorax, Vol: 53, ISSN: 0040-6376
Resolution of pulmonary oedema or the clearance of lung liquid depends on active sodium transport. A key component of this process is the sodium-potassium-adenosinetriphosphatase (Na+-K+-ATPase) enzyme, situated on the basolateral surface of alveolar epithelial cells, and known to be endogenously upregulated during oedema resolution. We have previously demonstrated that lung liquid clearance may be further enhanced by cationic liposome-mediated gene transfer of Na+K+ATPase cDNA in vivo to the lungs of a mouse model of high permeability pulmonary oedema induced by intraperitoneal injection of thiourea. In the following studies we have characterised the animal model and further assessed the effects of Na+-K+-ATPase gene transfer on the resolution of pulmonary oedema in vivo. Thus, intraperitoneal injection of thiourea (2.5 mg/kg) in C57/BL6 mice results in a time-dependent increase in lung liquid, as measured by lung wet weight:dry weight ratios, which peaks at 2 hours (6.3±0.3, n=8), is significantly (p<0.05) greater than untreated control mice (4.1±0.1, n=14) at this time point, and gradually declines over 24 hours (5.3±0.2, 5.1±0.1 and 4.5±0.1 at 4 hours, 6 hours and 24 hours respectively). Pretreatment of mice (24 hours prior to thiourea injection) by nasal sniffing of cationic liposome (lipid #67)-DNA complexes encoding the α and β subunits of Na+-K+-ATPase (160 μg/mouse), significantly (p<0.01) decreased the lung wet weight:dry weight ratios measured 2 hours after thiourea injection (5.1±0.1 n=15) compared to control animals (5.7±0.2, n=11) pretreated with an equivalent dose of an irrelevant gene (pCF1-CAT). The latter group was not significantly different compared to animals receiving no pretreatment (6.4±0.35, n=31, p>0.05). Nested RT-PCR on whole lung homogenates confirmed gene transfer by detection of vector-specific mRNA in 3 out of 4 mice studied 24 hours following gene transfer
Scheid P, Griesenbach U, Davies J, et al., 1998, Interleukin-8 production by airway epithelium and the effect of liposome-mediated IκB gene transfer, Thorax, Vol: 53, ISSN: 0040-6376
Interleukin-8 (IL-8) is a major chemoattractant for neutrophils, present in the CF lung in large numbers. We measured basal and stimulated IL-8 secretion from CF and non CF airway epithelial cell lines (CFTE and 16HBE cells, respectively) as well as from Cos7 cells in a controlled in vitro environment. Basal IL-8 secretion was significantly higher in CFTE cells (8110 ± 2286 pg/mg protein) than in 16HBE (1113 ± 123 pg/mg protein) and Cos7 (497 ± 138 pg/mg protein) [n=6, p<0.05]. Following TNF-α stimulation IL-8 secretion increased 8 ± 0.9 fold in CFTE, 2 ± 0.4 fold in 16HBE and 2.7 ± 0.2 fold in Cos7 cells. Preliminary data in freshly obtained human nasal cells also indicate an increased basal secretion of IL-8 (non CF: 62 ± 10 pg/ml n=16, CF: 165 ± 37 pg/ml n=12, p<0.05). The response to addition of Pseudomonas aeruginosa to these cells as measured by IL-8 secretion was also increased in CF (3279 ± 656 pg/ml n=10, p<0.05) when compared to non CF samples (817 ± 143 pg/ml n=8). Thus it appears that the IL-8 response is disproportionately greater in CF cells. It is likely that new treatment aimed at reducing IL-8 levels will be beneficial in CF. Nuclear factor kappa B (NFκB) is a transcription factor that regulates expression of inflammatory mediators including IL-8. NFκB is kept in an inactive state by association with a specific inhibitor (IκB). We investigated the effects of liposome-mediated overexpression of IκB in CFTE and Cos7 cells. Despite successful transfection, as confirmed by RT-PCR in both cell types, we did not detect an alteration of basal or TNF-α stimulated IL-8 secretion. This is most likely related to low levels of gene expression. In conclusion, we have shown that CF cells lines as well as CF primary cells produce increased levels of secreted IL-8 in the basal as well as in a stimulated state. Reduction of IL-8 secretion is likely to b
Munkonge FM, Hillery E, Griesenbach U, et al., 1998, Isolation of a putative nuclear import DNA shuttle protein, MOLECULAR BIOLOGY OF THE CELL, Vol: 9, Pages: 187A-187A, ISSN: 1059-1524
- Author Web Link
- Cite
- Citations: 3
Alton EWFW, Griesenbach U, Geddes DM, 1998, Milking gene therapy, NATURE MEDICINE, Vol: 4, Pages: 1121-1122, ISSN: 1078-8956
- Author Web Link
- Cite
- Citations: 5
Gosselin D, Stevenson MM, Cowley EA, et al., 1998, Impaired ability of <i>Cftr</i> knockout mice to control lung infection with <i>Pseudomonas aeruginosa</i>, AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, Vol: 157, Pages: 1253-1262, ISSN: 1073-449X
- Author Web Link
- Cite
- Citations: 66
Griesenbach U, Chonn A, Cassady R, et al., 1998, Comparison between intratracheal and intravenous administration of liposome-DNA complexes for cystic fibrosis lung gene therapy, GENE THERAPY, Vol: 5, Pages: 181-188, ISSN: 0969-7128
- Author Web Link
- Cite
- Citations: 68
Stern M, Caplen NJ, Browning JE, et al., 1998, The effect of mucolytic agents on gene transfer across a CF sputum barrier in vitro, GENE THERAPY, Vol: 5, Pages: 91-98, ISSN: 0969-7128
- Author Web Link
- Cite
- Citations: 90
Kent G, Iles R, Bear CE, et al., 1997, Lung disease in mice with cystic fibrosis, JOURNAL OF CLINICAL INVESTIGATION, Vol: 100, Pages: 3060-3069, ISSN: 0021-9738
- Author Web Link
- Cite
- Citations: 132
BAUMSTARKHAN C, GRIESENBACH U, RINK H, 1992, COMPARISON OF DNA STRAND BREAK INDUCTION IN CHO CELLS MEASURED BY ALKALINE ELUTION AND BY FLUOROMETRIC ANALYSIS OF DNA UNWINDING (FADU), FREE RADICAL RESEARCH COMMUNICATIONS, Vol: 16, Pages: 381-389, ISSN: 8755-0199
- Author Web Link
- Cite
- Citations: 15
, Paramyoxovirus Vector for Gene Transfer to the Cardiovascular System, TOKUGAN 2000-339942
This data is extracted from the Web of Science and reproduced under a licence from Thomson Reuters. You may not copy or re-distribute this data in whole or in part without the written consent of the Science business of Thomson Reuters.