Pyruvate carboxylase (PC) is a mitochondrial, biotin-containing enzyme catalyzing the ATP-dependent synthesis of oxaloacetate from pyruvate and bicarbonate, with a critical anaplerotic role in sustaining the brain metabolism. Based on the studies performed on animal models, PC expression was assigned to be glia-specific. To study PC distribution among human neural cells, we probed the cultured human astrocytes and brain sections with antibodies against PC. Additionally, we tested the importance of PC for the viability of cultured human astrocytes by applying the PC inhibitor 3-chloropropane-1,2-diol (CPD). Our results establish the expression of PC in mitochondria of human astrocytes in culture and brain tissue and also into a subpopulation of the neurons in situ. CPD negatively affected the viability of astrocytes in culture, which could be partially reversed by supplementing media with malate, 2-oxoglutarate, citrate, or pyruvate. The provided data estimates PC expression in human astrocytes and neurons in human brain parenchyma. Furthermore, the enzymatic activity of PC is vital for sustaining the viability of cultured astrocytes.

• MeSH
• astrocyty * metabolismus   MeSH
• kyselina pyrohroznová metabolismus   MeSH
• lidé MeSH
• mozek metabolismus   MeSH
• neurony metabolismus   MeSH
• pyruvátkarboxylasa * metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Pankreatický duktální adenokarcinom (PDAC) představuje jeden z nejagresivnějších typů lidských malignit. V současnosti je toto zhoubné onemocnění čtvrtou nejčastější příčinou úmrtí na rakovinu. Pětileté přežití pacientů s duktálním adenokarcinomem je méně než 8 %. Nové in vitro a in vivo modely jsou proto nutně potřebné pro vývoj nových terapií. S vlastní technologií pro derivaci nových, unikátních 3D nádorových buněčných linií izolovaných z lidských nádorů a ve spolupráci s Ústavem živočišné fyziologie a genetiky AV ČR v Liběchově představujeme plán vývoje velkého zvířecího modelu pro modelování lidského PDAC. S využitím tohoto modelu a nejmodernějších laboratorních technik provedeme profilovací analýzy (detekce a fenotypování cirkulujících rakovinných buněk, izolace a sekvenování cirkulující DNA, metabolomické profilování a analýzu onkoproteinů, a detekce cytokinů pomocí multiplexních protilátkových čipů za účelem nalezení biomarkerů nádoru pankreatu. Nově vyvinutý model zároveň poskytneme pro potřeby výzkumu spolupracujícím vědeckým pracovištím a farmaceutickým společnostem.

Pancreatic ductal adenocarcinoma represents one of the most aggressive type of human malignancy. Currently, this malignancy is the fourth most frequent cause of dead. 5-year survival of patients with ductal adenocarcinoma is less than 8 %. New in vitro and in vivo models are therefore desperately needed for new therapy development. With our own technology for derivation of new, unique 3D cancer cell lines from human tumors and in collaboration with Institute of Animal Physiology and Genetics, AS CR, in Libechov, we present here the plan for the development of large xenograft animal model (pig) bearing human pancreatic tumor. With the use of this model and state -of -the -art lab.techniques, we will perform multiple profiling analyses (circulating cancer cell detection and phenotyping, circulating DNA isolation and sequencing, metabolomic profiling and cancer -related proteins and cytokines detection with multiplex antibody array chips in order to find the hallmarks of pancreatic tumor.

Stromal-derived factor 1α (SDF‑1α, also known as CXCL12) is a chemokine that exerts its effects through the G-protein coupled receptors, C-X-C chemokine receptor type 4 (CXCR4) and 7 (CXCR7). There is marked evidence that the SDF-1/CXCR4 axis is involved in the pathogenesis of leukemia and therapies that target this axis are under development. The present study aimed to increase the efficacy of a DNA-based bcr-abl vaccine by simultaneously immunizing mice with a plasmid carrying the whole SDF-1α gene. Bcr-abl‑transformed 12B1 cells were used to challenge the mice. These cells have the oncogenic potential to induce both leukemia following intravenous inoculation and lymphoma-type solid tumors after subcutaneous inoculation. Administering an SDF‑1 carrying plasmid together with the bcr-abl vaccine resulted in increased survival following a challenge with subcutaneously administered 12B1 cells, although the difference was not statistically significant. However, there was a difference when the animals that developed subcutaneous tumors were only taken into consideration. In doubly-treated mice, significantly more mice failed to develop solid tumors than mice that had only received the bcr-abl vaccine. By contrast, the occurrence of fatal leukemia was significantly higher in the mice that were treated with the SDF-1 plasmid, regardless of whether they were immunized with the bcr-abl-vaccine. No humoral or cellular immune responses against SDF‑1 were detected in the treated mice, which suggested that the changes in oncogenic potential of 12B1 cells were due to the activity of SDF-1 itself.

• MeSH
• bcr-abl fúzní proteiny genetika   MeSH
• chemokin CXCL12 genetika   metabolismus   MeSH
• chronická myeloidní leukemie mortalita   terapie   veterinární   MeSH
• DNA vakcíny imunologie   terapeutické užití   MeSH
• faktor 1 indukovatelný hypoxií - podjednotka alfa metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• míra přežití MeSH
• molekulární sekvence - údaje MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• plazmidy genetika   metabolismus   MeSH
• receptory CXCR4 metabolismus   MeSH
• sekvence aminokyselin MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In the recent past, it has repeatedly been reported that CD4 cells play an important role in the immunology of chronic myeloid leukaemia. It was therefore of interest to test their activity in an animal model using bcr-abl-transformed cells. BALB/c mice were four times immunized with a DNA vaccine carrying the bcr-abl fusion gene. Two weeks after the last vaccine dose, the animals were challenged with syngeneic bcr-abl-transformed 12B1 cells which form solid tumors after subcutaneous administration. At the time of challenge, animals were treated with antibodies against the CD8+ T cells or CD4+ T cells. The efficacy of the depletion was monitored and found highly effective. All nonimmunized animals developed tumors. All animals untreated with the antibodies as well as those in which CD8+ T cells had been depleted, were fully protected against the challenge. On the other hand, almost all mice treated with anti-CD4+ antibody developed tumors. These results strongly suggested that the CD4+ T cells acted as effectors in the present system.

• MeSH
• antigeny CD95 imunologie   metabolismus   MeSH
• bcr-abl fúzní proteiny genetika   imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   metabolismus   MeSH
• CD8-pozitivní T-lymfocyty imunologie   metabolismus   MeSH
• DNA vakcíny imunologie   MeSH
• imunizace MeSH
• lidé MeSH
• ligand Fas metabolismus   MeSH
• lymfocytární deplece MeSH
• MHC antigeny II. třídy imunologie   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• nádorová transformace buněk genetika   imunologie   MeSH
• nádory genetika   imunologie   mortalita   prevence a kontrola   MeSH
• protinádorové vakcíny genetika   imunologie   MeSH
• slezina cytologie   imunologie   MeSH
• transformované buněčné linie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A series of DNA vaccines based on the bcr-abl fusion gene were developed and tested in mice. Two mouse (BALB/c) bcr-abl-transformed cell lines, B210 and 12B1, which both expressed p210bcr-abl and were oncogenic for syngeneic animals but differed in some other respects, were used as a model system. In the first series of experiments, plasmids carrying either the complete bcr-abl fusion gene or a fragment thereof coding for a 25-amino acid-long junction zone (bcr-abl25aa) linked with genes coding for a variety of immunostimulatory factors were used as the DNA vaccines. A plasmid carrying the complete bcr-abl gene was capable of inducing protection against challenge with either B210 or 12B1 cells. However, the DNA vaccines based on the gene fragment coding for p25aabcr-abl did not induce significant protection. To localize the immunizing epitopes on the p210bcr-abl protein, the whole fusion gene was split into nine overlapping fragments and these, individually or in various combinations, were used for immunization. Although none of the vaccines based on any single fragment provided potent protection, some combinations of these fragment-based vaccines were capable of eliciting protection comparable to that seen after immunization with the whole-gene vaccine. Surprisingly, a mixture of six fragment-vaccines was more immunogenic than the complete set of fragment DNA vaccines. To analyze this phenomenon, the three fragments missing from the hexavaccine were either individually or in various combinations mixed with the hexavaccine. The results obtained suggested that the product of the fragment coding for 197 amino acids forming the N-terminal of the BCR protein was involved in the decreased immunogenicity. However, further experiments are needed to clarify the point. Additional experiments revealed that all the important epitopes were located in the ABL portion of the p210bcr-abl protein. The livers, spleens and bone marrows of the successfully immunized animals were tested for the presence of bcr-abl-positive cells by RT-PCR. The results were negative, this suggesting that these animals were free of any residual disease.

• MeSH
• bcr-abl fúzní proteiny genetika   imunologie   MeSH
• časové faktory MeSH
• chronická myeloidní leukemie genetika   imunologie   patologie   prevence a kontrola   MeSH
• DNA vakcíny genetika   imunologie   MeSH
• HL-60 buňky MeSH
• imunizace MeSH
• lidé MeSH
• mapování epitopu MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• peptidové fragmenty imunologie   MeSH
• protinádorové vakcíny genetika   imunologie   MeSH
• transfekce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

Two mouse HPV16-transformed cell lines, viz. MK16 cells, which induce metastasizing tumors, and TC-1 cells, which induce non-metastasizing tumors were transduced with the gene for mouse endostatin. Two clones constitutively expressing endostatin were isolated from each of them. They were denoted ME3 and ME9, and TE2 and TE5, respectively. When inoculated into mice, ME3 cells were non-oncogenic. Nearly all mice inoculated with ME9 cells developed tumors, but considerably later than did the parental MK16 cells and metastasis formation was strongly reduced in these animals. On the other hand, TE2 and TE5 cells displayed oncogenic potential similar to that of the parental cells. To provide more information on these different effects of endostatin production, cell lysates of all six lines studied were tested for the content of 25 factors known to be involved in angiogenesis. The parental MK16 cells differed from the parental TC-1 cells and also from all endostatin producing sublines by a markedly higher production of interleukin 1alpha (IL-1alpha) and, to a lesser extent, by a higher production of several other factors tested. Additional experiments indicated that the suppression of the production of IL-1alpha by the parental MK16 caused by endostatin was due to an autocrine mechanism.

• MeSH
• autokrinní signalizace MeSH
• časové faktory MeSH
• endostatiny genetika   metabolismus   MeSH
• endoteliální buňky metabolismus   MeSH
• geny ras MeSH
• interleukin-1alfa metabolismus   MeSH
• interleukin-2 genetika   metabolismus   MeSH
• kultivační média speciální metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorová transformace buněk MeSH
• nádory plic genetika   imunologie   metabolismus   prevence a kontrola   sekundární   virologie   MeSH
• onkogenní proteiny virové genetika   MeSH
• Papillomavirus E7 - proteiny MeSH
• proliferace buněk MeSH
• represorové proteiny genetika   MeSH
• transdukce genetická MeSH
• transformované buněčné linie MeSH
• virová transformace buněk MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakty MeSH