BACKGROUND & AIMS: Chromosomal instability (CIN) is a carcinogenesis event that promotes metastasis and resistance to therapy by unclear mechanisms. Expression of the colon cancer-associated transcript 2 gene (CCAT2), which encodes a long noncoding RNA (lncRNA), associates with CIN, but little is known about how CCAT2 lncRNA regulates this cancer enabling characteristic. METHODS: We performed cytogenetic analysis of colorectal cancer (CRC) cell lines (HCT116, KM12C/SM, and HT29) overexpressing CCAT2 and colon organoids from C57BL/6N mice with the CCAT2 transgene and without (controls). CRC cells were also analyzed by immunofluorescence microscopy, γ-H2AX, and senescence assays. CCAT2 transgene and control mice were given azoxymethane and dextran sulfate sodium to induce colon tumors. We performed gene expression array and mass spectrometry to detect downstream targets of CCAT2 lncRNA. We characterized interactions between CCAT2 with downstream proteins using MS2 pull-down, RNA immunoprecipitation, and selective 2'-hydroxyl acylation analyzed by primer extension analyses. Downstream proteins were overexpressed in CRC cells and analyzed for CIN. Gene expression levels were measured in CRC and non-tumor tissues from 5 cohorts, comprising more than 900 patients. RESULTS: High expression of CCAT2 induced CIN in CRC cell lines and increased resistance to 5-fluorouracil and oxaliplatin. Mice that expressed the CCAT2 transgene developed chromosome abnormalities, and colon organoids derived from crypt cells of these mice had a higher percentage of chromosome abnormalities compared with organoids from control mice. The transgenic mice given azoxymethane and dextran sulfate sodium developed more and larger colon polyps than control mice given these agents. Microarray analysis and mass spectrometry indicated that expression of CCAT2 increased expression of genes involved in ribosome biogenesis and protein synthesis. CCAT2 lncRNA interacted directly with and stabilized BOP1 ribosomal biogenesis factor (BOP1). CCAT2 also increased expression of MYC, which activated expression of BOP1. Overexpression of BOP1 in CRC cell lines resulted in chromosomal missegregation errors, and increased colony formation, and invasiveness, whereas BOP1 knockdown reduced viability. BOP1 promoted CIN by increasing the active form of aurora kinase B, which regulates chromosomal segregation. BOP1 was overexpressed in polyp tissues from CCAT2 transgenic mice compared with healthy tissue. CCAT2 lncRNA and BOP1 mRNA or protein were all increased in microsatellite stable tumors (characterized by CIN), but not in tumors with microsatellite instability compared with nontumor tissues. Increased levels of CCAT2 lncRNA and BOP1 mRNA correlated with each other and with shorter survival times of patients. CONCLUSIONS: We found that overexpression of CCAT2 in colon cells promotes CIN and carcinogenesis by stabilizing and inducing expression of BOP1 an activator of aurora kinase B. Strategies to target this pathway might be developed for treatment of patients with microsatellite stable colorectal tumors.

• Klíčová slova
• Aneuploidy, MSS, Noncoding RNA, Tumorigenesis,
• MeSH
• Aurora kinasa B metabolismus   MeSH
• azoxymethan toxicita   MeSH
• chemorezistence genetika   MeSH
• chromozomální nestabilita * MeSH
• cytogenetické vyšetření MeSH
• dextrany toxicita   MeSH
• experimentální nádory chemicky indukované   genetika   patologie   MeSH
• genový knockdown MeSH
• karcinogeneze genetika   MeSH
• kolon cytologie   patologie   MeSH
• kolorektální nádory chemicky indukované   genetika   patologie   MeSH
• lidé MeSH
• myši transgenní MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• organoidy MeSH
• primární buněčná kultura MeSH
• proteiny vázající RNA genetika   metabolismus   MeSH
• protokoly protinádorové kombinované chemoterapie farmakologie   terapeutické užití   MeSH
• protoonkogenní proteiny c-myc metabolismus   MeSH
• regulace genové exprese u nádorů MeSH
• RNA dlouhá nekódující genetika   metabolismus   MeSH
• signální transdukce genetika   MeSH
• střevní sliznice cytologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• AURKB protein, human MeSH Prohlížeč
• Aurora kinasa B MeSH
• azoxymethan MeSH
• BOP1 protein, human MeSH Prohlížeč
• dextrany MeSH
• long non-coding RNA CCAT2, human MeSH Prohlížeč
• MYC protein, human MeSH Prohlížeč
• proteiny vázající RNA MeSH
• protoonkogenní proteiny c-myc MeSH
• RNA dlouhá nekódující MeSH

Standard-of-care chemo- or radio-therapy can induce, besides tumor cell death, also tumor cell senescence. While senescence is considered to be a principal barrier against tumorigenesis, senescent cells can survive in the organism for protracted periods of time and they can promote tumor development. Based on this emerging concept, we hypothesized that elimination of such potentially cancer-promoting senescent cells could offer a therapeutic benefit. To assess this possibility, here we first show that tumor growth of proliferating mouse TC-1 HPV-16-associated cancer cells in syngeneic mice becomes accelerated by co-administration of TC-1 or TRAMP-C2 prostate cancer cells made senescent by pre-treatment with the anti-cancer drug docetaxel, or lethally irradiated. Phenotypic analyses of tumor-explanted cells indicated that the observed acceleration of tumor growth was attributable to a protumorigenic environment created by the co-injected senescent and proliferating cancer cells rather than to escape of the docetaxel-treated cells from senescence. Notably, accelerated tumor growth was effectively inhibited by cell immunotherapy using irradiated TC-1 cells engineered to produce interleukin IL-12. Collectively, our data document that immunotherapy, such as the IL-12 treatment, can provide an effective strategy for elimination of the detrimental effects caused by bystander senescent tumor cells in vivo.

• Klíčová slova
• IL-12, cancer chemotherapy, cell therapy, cellular senescence, docetaxel,
• MeSH
• bystander efekt účinky léků   MeSH
• časové faktory MeSH
• cytokiny genetika   metabolismus   MeSH
• docetaxel MeSH
• experimentální nádory genetika   metabolismus   terapie   MeSH
• imunoterapie adoptivní metody   MeSH
• interleukin-12 biosyntéza   farmakologie   MeSH
• kombinovaná terapie MeSH
• myši inbrední C57BL MeSH
• nádorové buněčné linie MeSH
• protinádorové látky farmakologie   MeSH
• stárnutí buněk účinky léků   MeSH
• taxoidy farmakologie   MeSH
• tumor burden účinky léků   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny MeSH
• docetaxel MeSH
• interleukin-12 MeSH
• protinádorové látky MeSH
• taxoidy MeSH

To elucidate the immunological mechanisms critical for tumor progression, we bred novel mouse strains, different in the NKC and H-2D domains. We used inbreeding to generate hybrids of Balb/c and C57BL/6 of stable H-2Db+d-NK1.1neg and H-2Db-d+NK1.1high phenotypes. We analyzed the growth of three established MHC class I-deficient tumor cell lines: TC-1/A9 tumor (HPV-associated) and B16F10 melanoma, both syngeneic to C57BL/6, and the MCB8 (3-methycholanthrene-induced tumor) syngeneic to Balb/c. Furthermore, we induced colorectal carcinoma by azoxymethane-DSS treatment to test the susceptibility to chemically-induced primary cancer. We found that the novel strains spontaneously regressed the tumor transplants syngeneic to both Balb/c (MCB8) and C57BL/6 (B16F10 and TC-1/A9) mice. The H2-Db+d-NK1.1neg, but not the H2-Db-d+NK1.1high strain was also highly resistant to chemically-induced colorectal cancer in comparison to the parental mice. The immune changes during TC-1/A9 cancer development involved an increase of the NK cell distribution in the peripheral blood and spleen along with higher expression of NKG2D activation antigen; this was in correlation with the time-dependent rise of cytotoxic activity in comparison to C57BL/6 mice. The TC-1/A9 cancer regression was accompanied by higher proportion of B cells in the spleen and B220+/CD86+ activated antigen-presenting B cells distributed in the lymphoid organs, as well as in the periphery. The changes in the T-cell population were represented mainly by the prevalence of T helper cells reflected by grown CD4/CD8 ratio, most prominent in the b+d-NK1.1neg strain. The results of the present study imply usefulness of the two novel mouse strains as an experimental model for further studies of tumor resistance mechanisms.

• MeSH
• buňky NK imunologie   patologie   MeSH
• experimentální nádory genetika   imunologie   patologie   MeSH
• lektinové receptory NK-buněk - podrodina K biosyntéza   imunologie   MeSH
• lidé MeSH
• MHC antigeny I. třídy genetika   imunologie   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• regulace genové exprese u nádorů genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Klrk1 protein, mouse MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• MHC antigeny I. třídy MeSH

Recognition of glycosylation patterns is one of the basic features of innate immunity. Ability of C-type lectin-like receptors such as NKR-P1 to bind saccharide moieties has become recently a controversial issue. In the present study, binding assay with soluble fluorescently labeled recombinant rat NKR-P1A and mouse NKR-P1C proteins revealed apparently no affinity to the various neoglycoproteins. Lack of functional linkage between NKR-P1 and previously described saccharide binder was supported by the fact, that synthetic N-acetyl-D-glucosamine octabranched dendrimer on polyamidoamine scaffold (GN8P) did not change gene expression of NKR-P1 isoforms in C57BL/6 and BALB/c mice divergent in the NK gene complex (both in vitro and in vivo). Surprisingly, N-acetyl-D-glucosamine-coated tetrabranched polyamido-amine dendrimer specifically binds to NKT cells and macrophages but not to NK cells (consistently with changes in cytokine patterns). Despite the fact that GN8P has been tested as an immunomodulator in anti-cancer treatment animal models for many years, surprisingly no changes in cytokine profiles in serum relevant to anti-cancer responses using B16F10 and CT26 harboring mouse strains C57BL/6 and BALB/c are observed. Our results indicate possible indirect involvement of NK cells in GN8P mediated immune responses.

• Klíčová slova
• Anti-tumor immunity, C-type lectin related protein, Carbohydrate dendrimer, Clr, GN4P-A: GlcNAc4-PAMAM-ATTO 565, GN4P-NH(2)-GlcNAc(4)-PAMAM, GN4P: GlcNAc4-PAMAM, GN8P: GlcNAc8-PAMAM, GlcNAc, Gzmb, Macrophages, N-acetyl-d-glucosamine, N-acetyl-d-glucosamine-coated octabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer fluorescently labeled with ATTO 565, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer with free NH(2) group, NK cells, NKG2D, NKR-P1, NKR-P1 receptors, NKT cells, PAMAM dendrimer, PMA, Prf, SBA, SMC, granzyme B, natural killer group 2, member D, natural killer receptor protein 1, perforin, phorbol 12-myristate 13-acetate, polyamidoamine dendrimer, soybean agglutinin, spleen mononuclear cell,
• MeSH
• acetylglukosamin imunologie   metabolismus   MeSH
• buňky NK imunologie   metabolismus   MeSH
• dendrimery metabolismus   MeSH
• experimentální nádory farmakoterapie   genetika   imunologie   MeSH
• exprese genu účinky léků   imunologie   MeSH
• glykokonjugáty imunologie   metabolismus   farmakologie   MeSH
• interferon gama krev   genetika   imunologie   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lektinové receptory NK-buněk - podrodina B genetika   imunologie   metabolismus   MeSH
• lektiny typu C genetika   imunologie   metabolismus   MeSH
• makrofágy imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• NKT buňky imunologie   metabolismus   MeSH
• oligosacharidy imunologie   metabolismus   MeSH
• polyaminy imunologie   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein - isoformy genetika   imunologie   metabolismus   MeSH
• průtoková cytometrie MeSH
• slezina cytologie   imunologie   metabolismus   MeSH
• TNF-alfa krev   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylglukosamin MeSH
• dendrimery MeSH
• glykokonjugáty MeSH
• interferon gama MeSH
• lektinové receptory NK-buněk - podrodina B MeSH
• lektiny typu C MeSH
• oligosacharidy MeSH
• Poly(amidoamine) MeSH Prohlížeč
• polyaminy MeSH
• protein - isoformy MeSH
• TNF-alfa MeSH

Epigenetic events play an important role in tumour progression and also contribute to escape of the tumour from immune surveillance. In this study, we investigated the up-regulation of major histocompatibility complex (MHC) class I surface expression on tumour cells by epigenetic mechanisms using a murine tumour cell line expressing human E6 and E7 human papilloma virus 16 (HPV16) oncogenes and deficient in MHC class I expression, as a result of impaired antigen-presenting machinery (APM). Treatment of the cells with the histone deacetylase inhibitor Trichostatin A, either alone or in combination with the DNA demethylating agent 5-azacytidine, induced surface re-expression of MHC class I molecules. Consequently, the treated cells became susceptible to lysis by specific cytotoxic T lymphocytes. Further analysis revealed that epigenetic induction of MHC class I surface expression was associated with the up-regulation of APM genes [transporter associated with antigen processing 1 (TAP-1), TAP-2, low-molecular-mass protein 2 (LMP-2) and LMP-7]. The results demonstrate that expression of the genes involved in APM are modulated by epigenetic mechanisms and suggest that agents modifying DNA methylation and/or histone acetylation have the potential to change the effectiveness of antitumour immune responses and therapeutically may have an impact on immunological output.

• MeSH
• apoptóza účinky léků   MeSH
• azacytidin farmakologie   MeSH
• epigeneze genetická imunologie   MeSH
• experimentální nádory genetika   imunologie   virologie   MeSH
• geny MHC třídy I * MeSH
• histony metabolismus   MeSH
• infekce papilomavirem komplikace   MeSH
• inhibitory enzymů farmakologie   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• lidský papilomavirus 16 * MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buňky kultivované MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• prezentace antigenu genetika   imunologie   MeSH
• upregulace účinky léků   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azacytidin MeSH
• histony MeSH
• inhibitory enzymů MeSH
• kyseliny hydroxamové MeSH
• trichostatin A MeSH Prohlížeč

A better understanding of the molecular basis of tumor progression and invasion is needed to improve therapy for malignant tumors. Recently, we established a mouse metastatic MK16 model by transduction of secondary kidney cells with human papillomavirus type 16 (HPV16) E6 and E7 oncogenes and human H-ras activated by G12V mutation. In this study, we extended the model to MK16 cell lines derived from lung metastases and compared the oncogenicity of seven cell lines successively isolated from primary tumors or metastases. By observing the formation and growth of subcutaneous tumors and generation of lung metastasis, we showed a gradual increase in oncogenicity of MK16 cell lines. Interestingly, we demonstrated metastatic potential of MK16/A cells with low oncogenic potential in primary tumor development. To detect changes in gene expression associated with increasing oncogenicity of MK16 cell lines, we performed transcriptional profiling with the Atlas Plastic Mouse 5K microarray. We found that a substantial proportion of up-regulated genes encoded ribosomal proteins. Among the down-regulated genes, the highest number (n=10) belonged to a group coding for transcription factors. Expression of two of these, Pou3f2 and Gtl3, was reduced both in cells derived from primary tumors and those isolated from metastases. Furthermore, microarray hybridization suggested that the down-regulation of cyclin-dependent kinase inhibitors p16(Ink4a) and p57(Kip2) and up-regulation of A6 and A10 members of the S100 protein family might play a role in the increase of MK16 oncogenicity.

• MeSH
• experimentální nádory genetika   patologie   MeSH
• geny ras genetika   MeSH
• lidé MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorová transformace buněk genetika   MeSH
• nádorové buňky kultivované MeSH
• onkogenní proteiny virové genetika   MeSH
• Papillomavirus E7 - proteiny MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• progrese nemoci MeSH
• represorové proteiny genetika   MeSH
• reprodukovatelnost výsledků MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody   MeSH
• stanovení celkové genové exprese * MeSH
• transformované buněčné linie MeSH
• transplantace nádorů MeSH
• virová transformace buněk genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• E6 protein, Human papillomavirus type 16 MeSH Prohlížeč
• oncogene protein E7, Human papillomavirus type 16 MeSH Prohlížeč
• onkogenní proteiny virové MeSH
• Papillomavirus E7 - proteiny MeSH
• represorové proteiny MeSH

Escape of tumor cells from the host immune system is probably the most difficult obstacle to overcome in attempts to enhance the efficacy of immunotherapy of tumors. To solve the problem, animal models with escape mechanisms found in human tumors are needed. We have already established cell lines with substantially reduced expression of MHC class I molecules that were derived from oncogenic TC-1 cells producing E6 and E7 oncoproteins of HPV16. In this study, we prepared other cell lines from TC-1-induced tumors formed infrequently in mice immunized against the E7 antigen. These clones differed in morphology and both in vitro and in vivo growth properties. After vaccination with the highly efficient anti-E7 DNA vaccine several clones appeared to be quite resistant to induced anti-tumor immunity. Detection of the production of the MHC class I molecules and B7.1 costimulatory molecule did not provide clues to understand the mechanism of immunoresistance of the clones. Therefore, we performed transcriptional profiling using Atlas Mouse Cancer 1.2 Array (BD Clontech). Among the genes differently expressed in the examined cell lines were those of three cytokines influencing immune cells: MCP-1, osteopontin, and midkine. Altered secretion of MCP-1 chemokine was verified by ELISA. In addition, expression of the E7 oncogene was reduced in all TC-1 clones. Most importantly, one of anchor amino acids in the immunodominant epitope of E7 was mutated in all immunoresistant clones. Such mutations of HPV oncogenes must be considered in therapeutic-vaccine development and evaluation.

• MeSH
• antigeny CD80 genetika   metabolismus   MeSH
• buněčné klony MeSH
• buněčné linie MeSH
• chemokin CCL2 genetika   metabolismus   MeSH
• experimentální nádory genetika   imunologie   patologie   MeSH
• genetická transkripce genetika   MeSH
• messenger RNA genetika   metabolismus   MeSH
• metastázy nádorů MeSH
• MHC antigeny I. třídy genetika   metabolismus   MeSH
• mutace MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• onkogenní proteiny virové genetika   imunologie   MeSH
• Papillomavirus E7 - proteiny MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• proliferace buněk MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody   MeSH
• stanovení celkové genové exprese MeSH
• tvar buňky MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• antigeny CD80 MeSH
• Ccl2 protein, mouse MeSH Prohlížeč
• chemokin CCL2 MeSH
• messenger RNA MeSH
• MHC antigeny I. třídy MeSH
• oncogene protein E7, Human papillomavirus type 16 MeSH Prohlížeč
• onkogenní proteiny virové MeSH
• Papillomavirus E7 - proteiny MeSH

Groups of six BALB/c mice each were intravenously inoculated with lethal doses of Ba-P210 (B210) or 12B1 cells and examined by autopsy, histology, special staining methods, enzyme histochemistry and immunohistochemistry. Clinical symptoms related to neoplasia consisted of a poor nutritional state, anaemia, mild to moderate dehydration and apathy. Paresis was apparent in three mice inoculated with 12B1 cells. Necropsy revealed splenomegaly in all animals. Sporadic haemorrhages in the lungs and enlargement of some lymph nodes were seen in some of the animals. Histological examination showed neoplastic cells in the spleen, in the bone marrow of the sternum, in the lung interstitium and in sinusoids of the liver in all mice. In six of nine brains examined, mild to moderate infiltration by neoplastic cells was observed. In all but two mice mild infiltration of the kidneys was found. The enlargement of lymph nodes was caused by an accumulation of neoplastic cells. The paresis was due to neoplastic infiltration of the vertebra, epidural space and spinal roots. Staining with Sudan black revealed cytoplasmic granules in neoplastic cells; however, the peroxidase reaction was negative. Numerous neoplastic cells disseminated in the red pulp of the spleen were reactive with CD3, CD79beta, CD11b and with neutrophil antibodies. We classified the disease induced by both of the cell lines as acute myeloid undifferentiated leukaemia (AML MO).

• MeSH
• akutní nemoc MeSH
• endoteliální buňky patologie   MeSH
• experimentální nádory genetika   patologie   MeSH
• geny abl * MeSH
• imunohistochemie MeSH
• leukemická infiltrace MeSH
• myeloidní leukemie genetika   patologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nádorová transformace buněk genetika   MeSH
• páteř patologie   MeSH
• plíce patologie   MeSH
• progrese nemoci MeSH
• slezina imunologie   patologie   MeSH
• transformované buněčné linie * MeSH
• transplantace nádorů MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Recombinant vaccinia viruses derived from strain Praha, clone P13, and strain MVA were used for intratumoral delivery and expression of IL12 genes in tumors induced by HPV16 E6+E7 oncogenes in mice. Intratumoral injection of 10(3) PFU of P13-IL12 virus resulted in an increase of intra-tumoral IL12 on days 6-13, while only low levels of IL12 were found in sera. After the inoculation of 10(6) PFU of MVA-IL12, the same levels of IL12 were found as in animals injected with control virus. The intratumoral inoculation of 10(3) PFU P13-IL12 resulted in only approximately 30% of the tumors being virus positive, which was a consequence of reduced multiplication of the recombinant virus in vivo. The number of virus-positive tumors was not increased by repeated inoculations on three consecutive days. Intratumoral therapy with a dose of 10(3) PFU of P13-IL12 slowed down the growth of TC1 tumors, but never caused their regression. When local P13-IL12 treatment was combined with antigen-specific, DNA-vaccination therapy, no synergy between the two treatments was observed. The treatment with IL12-expressing virus retarded tumor growth to some degree, but did not change the number of regressing tumors. The highest efficacy of intra-tumoral P13-IL12 therapy was observed when the TC-1/A9 cell subline, with downregulated MHC class I expression, was used. TC-1/A9 tumors are less refractory to treatment with 10(3) P13-IL12/EL than are parental TC-1 cells.

• MeSH
• časové faktory MeSH
• DNA vakcíny imunologie   terapeutické užití   MeSH
• experimentální nádory genetika   imunologie   terapie   virologie   MeSH
• genetická terapie * MeSH
• genetické vektory genetika   MeSH
• Haplorrhini MeSH
• imunoterapie * MeSH
• infekce papilomavirem genetika   imunologie   terapie   MeSH
• interleukin-12 analýza   genetika   imunologie   terapeutické užití   MeSH
• lidé MeSH
• myši MeSH
• Papillomaviridae fyziologie   MeSH
• protinádorové vakcíny imunologie   terapeutické užití   MeSH
• transplantace nádorů MeSH
• virus vakcinie genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA vakcíny MeSH
• interleukin-12 MeSH
• protinádorové vakcíny MeSH

The effectiveness of chemoimmunotherapy with ifosfamide derivative CBM-4A and recombinant IL-2, IL-12, GM-CSF, or genetically modified, cytokine-producing tumour vaccines was examined in mice carrying HPV16-associated, MHC class I+ (TC-1), and MHC class I- (MK16) tumours. Intraperitoneal treatment of TC-1 or MK16 tumour-bearing mice with CBM-4A produced a significant tumour-inhibitory effect. When the i.p. treatment of the MHC class I+ TC-1 tumour-bearing mice with CBM-4A was followed by peritumoral s.c. administration of IL-2, IL-12, or both cytokines, the growth of TC1 tumours was inhibited more vigorously than after the chemotherapy alone. In contrast, when the i.p. treatment ofEthe MHC class I- MK16 tumour-bearing mice with CBM-4A was followed by peritumoral s.c. administration of IL-2 or IL-12, the cytokine therapy had no potentiating effect. The only potentiating effect of the MK16 tumour immunotherapy was obtained when the i.p. CBM-4A pretreatment was followed by peritumoral s.c. administration of IL-2 plus IL-12. InEfurther experiments, the TC-1 and MK16 tumour-bearing mice were i.p. pretreated with CBM-4A and then injected s.c., peritumorally, with genetically modified, IL-2 or GM-CSF-producing MK16 tumour vaccines. Whereas both genetically modified tumour vaccines produced a substantial tumour-inhibitory effect in mice carrying TC-1 tumours, no effect of the vaccines was observed in mice carrying MK16 tumour inocula. The systemic effects of local cytokine treatment were examined in mice carrying s.c. MK16 neoplasms, which were pretreated i.p. with CBM-4A, and then injected peritumorally with IL-2 or GM-CSF. Peritumoral administration of GM-CSF had no antimetastatic effect, whereas peritumoral IL-2 administration produced substantial reduction of lung metastases. The systemic antimetastatic effect of IL-2 contrasted with the negligible effect of IL-2 on the s.c. MK16 tumour inoculum. Taken collectively, the results indicate that in mice carrying the MK16 (MHC class I-) tumour, the effects of the adjuvant cytokine therapy were substantially weaker than in mice carrying the TC-1 (MHC class I+) tumour inoculum.

• MeSH
• adjuvancia imunologická aplikace a dávkování   terapeutické užití   MeSH
• alkylační protinádorové látky terapeutické užití   MeSH
• antigeny nádorové analýza   MeSH
• experimentální nádory genetika   imunologie   terapie   MeSH
• faktor stimulující granulocyto-makrofágové kolonie aplikace a dávkování   genetika   terapeutické užití   MeSH
• geny MHC třídy I MeSH
• geny ras MeSH
• ifosfamid analogy a deriváty   terapeutické užití   MeSH
• imunoterapie * MeSH
• injekce subkutánní MeSH
• interleukin-12 aplikace a dávkování   genetika   terapeutické užití   MeSH
• interleukin-2 aplikace a dávkování   genetika   terapeutické užití   MeSH
• kombinovaná terapie MeSH
• metastázy nádorů MeSH
• MHC antigeny I. třídy analýza   MeSH
• myši MeSH
• onkogenní proteiny virové genetika   fyziologie   MeSH
• Papillomavirus E7 - proteiny MeSH
• protinádorové vakcíny terapeutické užití   MeSH
• represorové proteiny * MeSH
• screeningové testy protinádorových léčiv MeSH
• vakcinace MeSH
• virová transformace buněk MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• adjuvancia imunologická MeSH
• alkylační protinádorové látky MeSH
• antigeny nádorové MeSH
• chlorobromofosfamide MeSH Prohlížeč
• E6 protein, Human papillomavirus type 16 MeSH Prohlížeč
• faktor stimulující granulocyto-makrofágové kolonie MeSH
• ifosfamid MeSH
• interleukin-12 MeSH
• interleukin-2 MeSH
• MHC antigeny I. třídy MeSH
• oncogene protein E7, Human papillomavirus type 16 MeSH Prohlížeč
• onkogenní proteiny virové MeSH
• Papillomavirus E7 - proteiny MeSH
• protinádorové vakcíny MeSH
• represorové proteiny * MeSH