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21.
In vivo cervical cancer growth inhibition by genetically engineered cytotoxic T cells 总被引:3,自引:0,他引:3
Dall P Herrmann I Durst B Stoff-Khalili MA Bauerschmitz G Hanstein B Niederacher D 《Cancer immunology, immunotherapy : CII》2005,54(1):51-60
Purpose: The CD44 v7/8 splice variant that is frequently expressed in cervical carcinoma and rarely expressed in normal tissues displays promising properties as a target antigen for cancer immune therapy. In this study, cytotoxic T lymphocytes (CTLs) were genetically engineered to gain CD44v7/8 target specificity. Methods: Clone 96 (CI96), an established murine cytotoxic T-cell line, and naïve murine T cells were retrovirally transduced with a fusion gene construct encoding for the single chain fragment scFv of the monoclonal antibody VFF17 and for the chain of the T-cell receptor (TCR). The therapeutic potential of genetically engineered T cells was tested in vitro and in vivo. Results: Surface expression of the chimeric TCR on infected Cl96 and naïve T cells was shown by FACS analysis. CD44v7/8-positive target cells were efficiently lysed by transduced Cl96 and naïve T cells, demonstrating the functionality and specificity of the chimeric TCR. In a xenograft BALB/c mouse model, efficient growth retardation of CD44v7/8-positive tumours was mediated by genetically engineered Cl96(VFF17)cyYZ cells. Conclusions: We were able to reprogramme the target specificity of recombinant Cl96 and naïve CTLs resulting in efficient cytolysis of CD44v7/8-positive cervical cancer cells. High transduction rates and the specific cytolysis of CD44v7/8-redirected CTLs are promising tools for an immune gene therapy approach for advanced cervical cancer.Abbreviations Ab
Antibody
- CTL
Cytolytic T lymphocyte
- mAb
Monoclonal antibody
- TCR
T-cell receptor 相似文献
22.
Deutsch GB Zielonka EM Coutandin D Weber TA Schäfer B Hannewald J Luh LM Durst FG Ibrahim M Hoffmann J Niesen FH Sentürk A Kunkel H Brutschy B Schleiff E Knapp S Acker-Palmer A Grez M McKeon F Dötsch V 《Cell》2011,144(4):566-576
TAp63α, a homolog of the p53 tumor suppressor, is a quality control factor in the female germline. Remarkably, already undamaged oocytes express high levels of the protein, suggesting that TAp63α's activity is under tight control of an inhibitory mechanism. Biochemical studies have proposed that inhibition requires the C-terminal transactivation inhibitory domain. However, the structural mechanism of TAp63α inhibition remains unknown. Here, we show that TAp63α is kept in an inactive dimeric state. We reveal that relief of inhibition leads to tetramer formation with ~20-fold higher DNA affinity. In vivo, phosphorylation-triggered tetramerization of TAp63α is not reversible by dephosphorylation. Furthermore, we show that a helix in the oligomerization domain of p63 is crucial for tetramer stabilization and competes with the transactivation domain for the same binding site. Our results demonstrate how TAp63α is inhibited by complex domain-domain interactions that provide the basis for regulating quality control in oocytes. 相似文献
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J P Salaun D Reichhart A Simon F Durst N O Reich P R Ortiz de Montellano 《Archives of biochemistry and biophysics》1984,232(1):1-7
Lauric acid in-chain hydroxylation is inhibited in microsomes from Jerusalem artichoke tubers (Helianthus tuberosus L.) incubated with 9-decenoic, 11-dodecenoic, or 11-dodecynoic acids. 9-Decenoic acid is at best a weak competitive inhibitor of the in-chain hydroxylase, but inactivates the enzyme in a time-dependent, pseudo-first-order process with a rate constant of approximately 1.1 X 10(-3) s-1. In contrast, 11-dodecenoic acid causes a slower, time-dependent loss of the hydroxylase activity, but is a potent competitive inhibitor of the enzyme (Ki = 2 microM). Neither agent decreases the microsomal concentration of cytochrome b5, NADH-cytochrome b5 reductase, or NADPH cytochrome P-450 reductase. Cinnamic acid 4-hydroxylation, catalyzed by a cytochrome P-450 enzyme, is not affected by concentrations of 9-decenoic acid that suppress lauric acid hydroxylation. 11-Dodecenoic acid is much less specific and, at higher concentrations, markedly reduces the microsomal cytochrome P-450 content, and the hydroxylation of both lauric and cinnamic acids. 相似文献
26.
Eliza I. Gilbert Scott L. Durst Angela P. James Jason E. Davis Thomas B. Sinclair Nathan R. Franssen 《Environmental Biology of Fishes》2018,101(6):1067-1076
Over evolutionary time, predator-prey interactions have shaped and constrained functional and behavioral traits of piscivorous fishes. The endangered Colorado Pikeminnow Ptychocheilus lucius, a large endemic piscivore of the Colorado River Basin, encounters a substantially altered prey base that differs in behaviors and morphologies compared to the historical suite of native prey. To assess physical limitations of Colorado Pikeminnow predation, we conducted a feeding experiment with two species of nonnative prey (spined and despined Channel Catfish Ictalurus punctatus and Red Shiner Cyprinella lutrensis) and quantified scaling of cranial morphology in this predator. In our predation experiments, Colorado Pikeminnow (215–312 mm total length) consumed both spined and despined Channel Catfish as well as Red Shiner but only consumed prey less than 20% of the predator’s total length. Previous feeding trials using smaller Colorado Pikeminnow, with native and nonnative prey species, indicated they consumed prey up to 35% of their total length, suggesting relative prey size limits may decrease as this predator grows. Morphological measurements also suggested relative prey size suitability may decrease as Colorado Pikeminnow become larger, with head depth and width demonstrating isometric scaling at small sizes and shifting to negative allometry as fish get larger. Together, these data suggest an ontogenetic shift in the head morphology of Colorado Pikeminnow may decrease the relative size of prey available to these predators. In severely altered systems, understanding trophic characteristics that limit overall predator resource availability will be critical for conservation of piscivorous fishes. 相似文献
27.
Grass was field-dried to 3 different dry matter (DM) levels (200, 430 and 540 g/kg) and inoculated with 106–107 cfu/g of a Listeria monocytogenes strain sharing a phagovar occasionally involved in food-borne outbreaks of listeriosis. Formic acid (3 ml/kg) or lactic acid
bacteria (8·105/g) with cellulolytic enzymes were applied only to forages with low and intermediate DM levels. Forages were ensiled in laboratory
silos (1700 ml) and were stored at 25°C for 30 or 90 days. After 90 days of storage, L. monocytogenes could not be detected in any silo, except one with the high dry matter grass without additive. After 30 days of storage,
between 102 and 106 cfu L. monocytogenes/g silage were isolated from the untreated silages. Increasing the DM content from 200 to 540 g/kg did not reduce listeria
counts possibly because of the lower production of fermentation acids (higher pH). In silages treated with additives, counts
of L. monocytogenes were always lower than in silages without additive. In wet silages (DM 200 g/kg) both additives were effective, but in the
wilted silages (DM 430 g/kg) only the bacterial additive reduced listeria counts below detection level. Listeria counts were
highly correlated to silage pH (r = 0.92), the concentration of lactic acid (r = -0.80) and the pooled amount of undissociated
acids (r = -0.83). 相似文献
28.
Purification and immunocharacterization of a plant cytochrome P450: the cinnamic acid 4-hydroxylase 总被引:9,自引:0,他引:9
B Gabriac D Werck-Reichhart H Teutsch F Durst 《Archives of biochemistry and biophysics》1991,288(1):302-309
Cinnamic acid 4-hydroxylase (CA4H) was purified from microsomes of manganese-induced Jerusalem artichoke (Helianthus tuberosus L.) tuber tissues. The three-step purification procedure involved solubilization and phase partitioning in Triton X-114, followed by chromatography on DEAE-Trisacryl and hydroxylapatite columns. Purification was monitored using carbon monoxide and type I substrate binding properties of the enzyme. The protein, purified to electrophoretic homogeneity, showed an Mr of about 57,000 on SDS-PAGE. Polyclonal antibodies raised against this protein selectively reacted with a 57-kDa polypeptide on Western blots of induced Jerusalem artichoke microsomes. The antibody selectively and strongly inhibited CA4H activity from several plant species. 相似文献
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Muhammad Asim Helmi Hussien Luis Poveda John Thor Arnason Tony Durst 《Phytochemistry》2010,71(11-12):1418-1422
Five spirocaracolitone triterpenoids were isolated from the dichloromethane-soluble fraction of the bark of Ruptiliocarpon caracolito, and their structures were determined mainly by application of 1D and 2D NMR spectroscopy. Two known CD-spirotriterpenoids were also isolated from the same source. This brings the total number of known CD-spirotriterpenoids from this source to 17. The discovery of such a large number of closely-related compounds in a single species represents an excellent example of phytochemical redundancy as a defense mechanism. 相似文献