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The tetracycline (Tet) system is widely used for regulation of gene expression in vitro and in vivo. We constructed C57BL/6 transgenic mice (rtTA-CM2) with strong and ubiquitous reverse transactivator (rtTA2(S)-M2) gene expression. rtTA-CM2 mice were crossed to Tet-responsive reporter mice (LC-1) conditionally expressing the firefly luciferase (FLuc) gene under control of a Tet-responsive element, which allowed sensitive quantification of the transactivator activity by bioluminescent imaging. Following doxycycline (dox) application, up to 10(5)-fold increase in BL signal was measured. rtTA activity was inducible in most analyzed organs. After dox withdrawal the BL signal decreased significantly but did not disappear completely, most likely due to a dox depot formation in vivo. The residual dox was sufficient to partly down-regulate a Tet-off controlled oncogene in a tumor transplantation experiment, resulting in reduced tumor growth. rtTA-CM2 mice may be a useful tool to analyze the function of genes in various organs but also reveal that down-regulation of gene expression is not complete. 相似文献
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Both soluble and membrane-bound forms of Flt3 ligand enhance tumor immunity following “suicide” gene therapy in a murine colon carcinoma model 总被引:2,自引:0,他引:2
Alsheikhly AR Zweiri J Walmesley AJ Watson AJ Christmas SE 《Cancer immunology, immunotherapy : CII》2004,53(11):946-954
In prodrug-activated (suicide) gene therapy, tumor cells are transfected with the gene for an enzyme that converts an inactive prodrug, such as ganciclovir (GCV), to a toxic compound. Transfected cells are killed on administration of GCV, as also are untransfected bystander cells. The ability of the dendritic cell stimulatory cytokine Flt3 ligand (Flt3-L) to modulate prodrug-activated gene therapy has been investigated. Transfectants of the murine colon carcinoma MC26 were generated expressing soluble (FLS) and membrane-bound forms of Flt3-L. They were inoculated together with wild-type MC26 cells and cells expressing herpes simplex virus-1 (HSV1) thymidine kinase into BALB/c mice, which were then administered GCV. Expression of Flt3-L or FLS prevented regrowth of tumor in most mice, which was comparable to the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF), while tumors recurred in all mice receiving suicide gene therapy alone. Recurring tumor cells were resistant to direct killing by GCV but sensitive to bystander killing in vitro. Mice without tumor recurrence were rechallenged with unmodified MC26 cells. Of those mice given transfectants expressing GM-CSF, Flt3-L, or FLS, approximately 50% were immune to rechallenge. These mice also showed cytotoxic and proliferative responses to MC26 cells. These experiments show that both soluble and membrane-bound forms of Flt3-L were able to induce a protective immune response to colon carcinoma cells in a fashion similar to GM-CSF.Abdul-Razzak Alsheikhly and Jehad Zweiri contributed equally to this work and are listed in alphabetical order. 相似文献
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Background
Colorectal carcinoma is a common and often fatal disease in which methods of early detection and monitoring are essential. The present study was conducted for measuring serum levels of nucleosomes, carcinoembryonic antigen (CEA) and CA 19-9 in patients newly diagnosed with colorectal carcinoma and confirmed by clinicopathological study. 相似文献6.
Carbon export from arbuscular mycorrhizal roots involves the translocation of carbohydrate as well as lipid 总被引:4,自引:0,他引:4
Bago B Pfeffer PE Abubaker J Jun J Allen JW Brouillette J Douds DD Lammers PJ Shachar-Hill Y 《Plant physiology》2003,131(3):1496-1507
Arbuscular mycorrhizal (AM) fungi take up photosynthetically fixed carbon from plant roots and translocate it to their external mycelium. Previous experiments have shown that fungal lipid synthesized from carbohydrate in the root is one form of exported carbon. In this study, an analysis of the labeling in storage and structural carbohydrates after (13)C(1) glucose was provided to AM roots shows that this is not the only pathway for the flow of carbon from the intraradical to the extraradical mycelium (ERM). Labeling patterns in glycogen, chitin, and trehalose during the development of the symbiosis are consistent with a significant flux of exported glycogen. The identification, among expressed genes, of putative sequences for glycogen synthase, glycogen branching enzyme, chitin synthase, and for the first enzyme in chitin synthesis (glutamine fructose-6-phosphate aminotransferase) is reported. The results of quantifying glycogen synthase gene expression within mycorrhizal roots, germinating spores, and ERM are consistent with labeling observations using (13)C-labeled acetate and glycerol, both of which indicate that glycogen is synthesized by the fungus in germinating spores and during symbiosis. Implications of the labeling analyses and gene sequences for the regulation of carbohydrate metabolism are discussed, and a 4-fold role for glycogen in the AM symbiosis is proposed: sequestration of hexose taken from the host, long-term storage in spores, translocation from intraradical mycelium to ERM, and buffering of intracellular hexose levels throughout the life cycle. 相似文献
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Effects of nitrogen supply on growth, yield and yield components of safflower and sunflower 总被引:2,自引:0,他引:2
Safflower (Carthamus tinctorius L.) represents an important oil crop internationally and may have a certain production potential under low input conditions,
particularly in organic farming systems, where the putatively low nutrient requirement would be an advantage. However, little
is known about the nutrient requirements of safflower. This study was undertaken to determine the growth and yield response
of safflower as compared to sunflower (Helianthus annuus L.) under different N supplies. Safflower and sunflower plants were grown in Mitscherlich pots containing equal volumes of
sand, nutrient-poor limed soil, and perlite. Nitrogen supply was the same for both species (0.25, 0.5, 1.0, 1.5, and 2.0 g
per pot) in the first year, but was raised for sunflower (0.5, 1.0, 2.0, 3.0, and 4.0 g per pot) in the second. Increased
N supply enhanced plant growth and yield for both species. Growth and yield of safflower increased up to 1.0 g pot−1, while the optimum for sunflower was 2.0 g pot−1. Safflower out-yielded sunflower at low N supply, while at high N level the opposite occurred. Functional analysis according
to Michaelis–Menten revealed that – in terms of yield formation – safflower is superior to sunflower under N-limited conditions.
Safflower is generally more efficient than sunflower in concentrating N in their shoots. N concentration in the photosynthetically
active youngest mature blade of both species exhibited the same functional relationship with the N supply, but safflower required
a much lower leaf N concentration to produce optimal yield as compared to sunflower, indicating the higher efficiency of the
former in terms of NUE. Yield components analysis revealed that in safflower, yield is tightly correlated with the number
of capitula per plant and the mass per achene, both being strongly correlated, too. On the contrary, sunflower yield was merely
determined by the number of achenes per capitulum, followed by the mass per achene. Path coefficient analysis showed that
in safflower, the direct effects of the achene and leaf N content as well as the leaf dry matter on oil yield are small, and
mediated principally via indirect effects on the number of achenes per capitulum, while for sunflower the number of achenes
per capitulum exerts a strong direct effect.
Dedicated to the late Prof. Dr. Burkhard Sattelmacher, who passed away on November 21, 2005. 相似文献
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Vertuani S Triulzi C Roos AK Charo J Norell H Lemonnier F Pisa P Seliger B Kiessling R 《Cancer immunology, immunotherapy : CII》2009,58(5):653-664
To study DNA vaccination directed against human HER-2 in the HHD mouse Tg strain, we created a novel HER-2-expressing syngeneic
tumor transplantation model. We found that a DNA vaccine encoding the full length HER-2 DNA protected HHD mice from HER-2+ tumor challenge by a CTL independent mechanism. A more efficient approach to induce HLA-A2 restricted CTLs, through immunization
with a multi-epitope DNA vaccine expressing the HLA-A2 restricted HER-2 369–377, 435–443 and 689–697 epitopes, resulted in
high numbers of peptide specific T cells but failed to induce tumor protection. Subsequently we discovered that HER-2 transfected
tumor cells down-regulated MHC class I antigen expression and exhibited a series of defects in the antigen processing pathway
which impaired the capacity to produce and display MHC class I peptide-ligands to specific CTLs. Our data demonstrate that
HER-2 transfection is associated with defects in the MHC class I presentation pathway, which may be the underlying mechanism
behind the inability of CTLs to recognize tumors in this HLA-A2 transgenic model. As defective MHC class I presentation may
be a common characteristic of HER-2 expressing tumors, vaccines targeting HER-2 should aim at inducing an integrated immune
response where also CD4+ T cells and antibodies are important components.
S. Vertuani and C. Triulzi contributed equally to this work. 相似文献
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Kurte M López M Aguirre A Escobar A Aguillón JC Charo J Larsen CG Kiessling R Salazar-Onfray F 《Journal of immunology (Baltimore, Md. : 1950)》2004,173(3):1731-1737
Tumor cells treated with IL-10 were shown to have decreased, but peptide-inducible expression of MHC class I, decreased sensitivity to MHC class I-restricted CTL, and increased NK sensitivity. These findings could be explained, at least partially, by a down-regulation of TAP1/TAP2 expression. In this study, IT9302, a nanomeric peptide (AYMTMKIRN), homologous to the C-terminal of the human IL-10 sequence, was demonstrated to mimic these previously described IL-10 effects on MHC class I-related molecules and functions. We observed a dose-dependent down-regulation of MHC class I at the cell surface of melanoma cells after 24-h treatment with IT9302. The IL-10 homologue peptide also caused a dose-dependent inhibition of the IFN-gamma-mediated surface induction of MHC class I in a melanoma cell line. We demonstrated, using Western blot and flow cytometry, that IT9302 inhibits the expression of TAP1 and TAP2 proteins, but not MHC class I H chain or low molecular protein molecules. Finally, peptide-treated melanoma cells were shown to be more sensitive to lysis by NK cells in a dose-dependent way. Taken together, these results demonstrate that a small synthetic peptide derived from IL-10 can mimic the Ag presentation-related effects mediated by this cytokine in human melanomas and increase tumor sensitivity to NK cells, which can be relevant in the designing of future strategies for cancer immune therapy. 相似文献
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Gingerenone A Attenuates Monocyte‐Endothelial Adhesion via Suppression of I Kappa B Kinase Phosphorylation 下载免费PDF全文
Hee Joo Kim Joe Eun Son Jae Hwan Kim Charles C. Lee Hee Yang Soonham Sami Yaghmoor Youssri Ahmed Jehad Mustafa Yousef Khalid Omer Abualnaja Abdulrahman Labeed Al‐Malki Taha Abdullah Kumosani Jong Hun Kim Jung Han Yoon Park Chang Yong Lee Jong‐Eun Kim Ki Won Lee 《Journal of cellular biochemistry》2018,119(1):260-268