RESUMEN
OBJECTIVE: To investigate the utility of asymptomatic screening, including CA-125, imaging, and pelvic exam, in the diagnosis and management of recurrent ovarian cancer. METHODS: Women with ovarian cancer whose cancer recurred after remission were categorized by first method that their provider suspected disease recurrence: CA-125, imaging, symptoms, or physical exam. Differences in clinicopathologic, primary treatment characteristics, and outcomes data including secondary cytoreductive surgery (SCS) outcome and overall survival (OS) were collected. RESULTS: 102 patients were identified at our institution from 2003 to 2015. 20 recurrences were detected by symptoms, while 62 recurrences were diagnosed first by asymptomatic rise in CA-125, 5 by pelvic exam, and 15 by imaging in the absence of known exam abnormality or rise in CA-125.Mean time to recurrence was 18.9 months, and median survival was 45.8 months. These did not vary by recurrence detection method (all p > 0.4). Patients whose disease was detected by CA-125 were less likely to undergo SCS than those detected by other means (21.7% vs. 35.0%, p = 0.007). In addition to the 5 patients whose recurrence was detected primarily by pelvic exam, an additional 10 (total n = 15) patients had an abnormal pelvic exam at time of diagnosis of recurrence. DISCUSSION: Recurrence detection method was not associated with differing rates of survival or optimal SCS, however those patients detected by CA-125 were less likely to undergo SCS. The pelvic exam was a useful tool for detecting a significant proportion of recurrences.
RESUMEN
BACKGROUND: Extramammary Paget disease (EMPD) is often associated with underlying or distant synchronous malignancies. The prognosis for affected patients is generally favourable; however, the risk of secondary malignancies is unknown. OBJECTIVES: The goal of the study was to analyse the incidence, prognosis and pattern of secondary malignancies for patients with invasive EMPD using data from the Surveillance, Epidemiology and End Results (SEER) Program. METHODS: We searched the SEER Program database for patients diagnosed with invasive EMPD between 1973 and 2008. Demographic data, outcome and secondary malignancies more than 1 year after the initial diagnosis of invasive EMPD were included in the analysis. We calculated the standardized incidence ratio (SIR) and estimated the excess absolute risk (EAR) per 10,000 person-years (PY). RESULTS: There were 1439 patients who were diagnosed with invasive EMPD. Most patients (80.4%) had localized disease, while 17.1% had locoregional spread and 2.5% presented with distant disease. The SIR for secondary malignancies in patients with invasive EMPD was significantly elevated with an EAR of 97.4 additional malignancies per 10,000 PY. The excess risk was mostly due to a significantly increased incidence of colorectal and anal malignancies. The initial site of disease predicted the site of the secondary malignancies, with patients with colorectal, anal, vulvar and scrotal disease showing an increased risk of colorectal, anal, vulvar and scrotal malignancies, respectively. CONCLUSIONS: Our study identified a long-term increased risk of developing secondary malignancies in patients with invasive EMPD that are mainly related to the site of origin of this disease. Patients with invasive EMPD require prolonged follow-up and screening for these malignancies.
Asunto(s)
Neoplasias Primarias Secundarias/epidemiología , Enfermedad de Paget Extramamaria/epidemiología , Anciano , Neoplasias del Ano/epidemiología , Neoplasias Colorrectales/epidemiología , Femenino , Neoplasias de los Genitales Masculinos/epidemiología , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Pronóstico , Factores de Riesgo , Escroto , Neoplasias Cutáneas/epidemiología , Neoplasias de la Vulva/epidemiologíaRESUMEN
Duboisia sp. is a small tree belonging to the family Solanaceae originating from the rainforest areas of the eastern coast of Australia. Dried leaves are used for the extraction of pharmaceutical alkaloids, making this a commercially viable crop. The root-knot nematode Meloidogyne incognita has been reported parasitizing Duboisia myoporoides (5); however, no information of other root-knot nematode species associated with this plant was found. Duboisia sp. is cultivated at Solana Farm, near Arapongas (23°25'08â³ S, 51°25'26â³ W), Paraná State, Brazil. During the renovation of a production field in this municipality, galled roots were observed on plants and samples were submitted to the Nematology Laboratory at Instituto Agronômico do Paraná, IAPAR, on December 2013. Plants did not exhibit any above-ground symptoms. The specimens were identified through perineal patterns and esterase phenotypes of 20 adult females extracted from dissected roots (2,3) and morphometrics of 10 second-stage juveniles extracted from roots using the blender-sieving method (1). Morphological characteristics were consistent with those described for M. javanica (4). Females had rounded perineal patterns with low, trapezoid shape dorsal arch, striae smooth interrupted by a pair of incisures on both sides, corresponding to lateral fields, clearly demarcated from striae by more or less parallel lines, tail whorl often distinct (4). The juvenile mean body length was 459.9 ± 28.7 µm and tail length averaged 51.6 ± 5.1 µm, with 10 to 16 µm long hyaline region and finely rounded tail tip (4). Results from the esterase electrophoresis were typical of M. javanica (2) with the J3 (Rm = 1.0, 1.3, and 1.4) phenotype being obtained. To our knowledge, this is the first report of M. javanica on Duboisia sp. in Brazil. This finding has great importance for Brazilian production since this nematode may damage plants, reduce yields, and control of this nematode on Duboisia sp. is difficult (5). Additional work is necessary in order to elucidate the losses caused by M. javanica on Duboisia sp. References: (1) J. I. Bonetti and S. Ferraz. Fitopatol. Bras. 6:533, 1981. (2) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 22:10, 1990. (3) K. M. Hartman and J. N. Sasser. Page 115 in: An Advanced Treatise on Meloidogyne. Volume II Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh, 1985. (4) D. J. Hunt and Z. A. Handoo. Page 55 in: Root-Knot Nematodes. R. N. Perry et al., eds. CABI International, Wallingford, UK, 2010. (5) A. M. Mello et al. Nematol. Bras. 22(2):12, 1998.
RESUMEN
Common bean (Phaseolus vulgaris F.) is one of the most important crops in Paraná State, which is responsible for almost 10% of the Brazilian production (4). Root knot nematodes, Meloidogyne spp., are common parasites of this crop worldwide, but damage caused by Meloidogyne inornata has not been reported. During a survey of nematode species present on common bean fields in Paraná State, Brazil, galled root samples of cultivars Tuiuiú and Eldorado were submitted, in June 2012, in the Nematology Laboratory from IAPAR, collected in the municipalities of Araucária (25°35'34â³S, 49°24'36â³W) and Santana do Itararé (23°45'18â³S, 49°37'44â³W). Plants did not exhibit any above-ground symptoms. The specimens were identified through perineal patterns and esterase phenotypes of 20 adult females extracted from dissected roots (2,3). The population densities observed in the samples were 140 and 700 J2 and eggs per gram of roots, respectively, for both samples. Characteristics were consistent with those described for M. inornata. For example, perineal patterns of M. inornata showed a high dorsal arch, with smooth to wavy striae, similar to those of M. incognita; but no punctate markings between anus and tail terminus were observed. However, from the esterase electrophoresis we obtained the I3 (Rm = 0.83, 1.15, and 1.32) phenotype, typical of M. inornata, a species-specific phenotype used to differentiate this species from M. incognita (1). Moreover, the excretory pore of adult females was located 32.1 (± 5.4) µm from the anterior end, consistent with the M. inornata description (25 to 53 µm) (1). To the best of our knowledge, this is the first report of M. inornata parasitizing common bean roots. This finding has great importance for Brazilian agriculture, since this nematode may damage common bean plants and become an additional problem for this crop. Additional work is necessary in order to elucidate the losses caused by M. inornata on common bean. References: (1) R. M. D. G. Carneiro et al. Nematology 10:123, 2008. (2) P. R. Esbenshade and A. C. Triantaphyllou J. Nematol. 22:10, 1990. (3) K. M. Hartman and J. N. Sasser. Page 115 in: An Advanced Treatise on Meloidogyne, Volume II Methodology. K. R. Barker et al., eds. Raleigh: North Carolina State University Graphics, 1985. (4) MAPA. Feijão, Ministério da Agricultura, Brasil. Retrieved from http://www.agricultura.gov.br/vegetal/culturas/feijao September 05, 2012.
RESUMEN
Posttransplant malignancy developing in an allograft is an uncommon complication of organ transplantation. The tumor may represent malignant transformation of donor or recipient cells that were previously normal, metastatic malignancy of recipient origin or malignancy transmitted from organ donor to recipient. Establishing the origin of the malignancy is critical to treatment algorithms. It is generally believed allograft removal and immunosuppression withdrawal will lead to resolution of transmitted malignancies in cases where the renal allograft is the origin. We report a male patient who developed metastatic ovarian malignancy secondary to donor transmission.
Asunto(s)
Adenocarcinoma Mucinoso/etiología , Neoplasias Renales/etiología , Trasplante de Riñón/efectos adversos , Neoplasias Ováricas/etiología , Donantes de Tejidos , Adenocarcinoma Mucinoso/secundario , Adulto , Resultado Fatal , Femenino , Humanos , Neoplasias Renales/secundario , Neoplasias Hepáticas/secundario , Metástasis Linfática , Masculino , Persona de Mediana Edad , Nefrectomía , Neoplasias Ováricas/patología , Tomografía Computarizada por Rayos XRESUMEN
We report the immunological characterization of three colon carcinoma cell lines, COLO 205, SW620 and SW403, which we selected to combine with cytokine-secreting fibroblasts for the development of an allogeneic tumour cell vaccine. The cell lines expressed HLA-A2 as well as shared tumour-associated antigens (TAAs) representative of colon carcinomas: CEA, Ep-CAM, MUC1, HER2/neu and MAGE antigens. They did not secrete high levels of the immunosuppressive factors TGF-beta, IL-10 or prostaglandins. The lines presented TAAs in a manner recognized by immune effector cells, which was demonstrated by the lysis of SW620 by HLA-A2-restricted anti-p53 cytotoxic T lymphocytes (CTL). COLO 205 and SW620 were genetically modified to express the co-stimulatory molecule CD80 (B7.1), which increased the ability of the cells to stimulate CTL in vitro. CTL clones derived from HLA-A2+ peripheral blood mononuclear cells stimulated with the CD80-expressing lines lysed the stimulator cell and an HLA-A2+ colon cancer cell line, but did not lyse an isogeneic fibroblast line or an HLA-A2- colon cancer cell line. CTL clones derived from colon carcinoma patients immunized with an allogeneic vaccine containing these lines demonstrated killing of autologous tumour cells, the vaccine cell lines and other HLA-A2+ colon cancer cell lines, but not fibroblasts isogeneic to certain of the target cell lines. Our studies demonstrate that these colon carcinoma cell lines express shared TAAs that can induce CTLs which recognize and lyse other colon carcinoma cells, and support the continued clinical evaluation of the CD80 gene modified allogeneic colon cell/cytokine-secreting fibroblast carcinoma vaccine.
Asunto(s)
Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Neoplasias del Colon/inmunología , Antígeno HLA-A2/inmunología , Isoantígenos/inmunología , Células Tumorales Cultivadas/inmunología , Presentación de Antígeno , Antígeno B7-1/genética , Antígeno B7-1/inmunología , Antígeno Carcinoembrionario/inmunología , Moléculas de Adhesión Celular/inmunología , Células Cultivadas , Neoplasias del Colon/prevención & control , Citocinas/metabolismo , Citotoxicidad Inmunológica , Molécula de Adhesión Celular Epitelial , Fibroblastos/inmunología , Fibroblastos/metabolismo , Humanos , Activación de Linfocitos , Mucina-1/inmunología , Proteínas de Neoplasias/inmunología , Receptor ErbB-2/inmunología , Proteínas Recombinantes de Fusión/inmunología , Linfocitos T Citotóxicos/inmunología , Factor de Crecimiento Transformador beta/metabolismoAsunto(s)
Obstrucción Intestinal/cirugía , Leiomiomatosis/cirugía , Neoplasias Peritoneales/cirugía , Enfermedades del Sigmoide/cirugía , Neoplasias del Colon/complicaciones , Neoplasias del Colon/cirugía , Femenino , Estudios de Seguimiento , Humanos , Neoplasias del Íleon/complicaciones , Neoplasias del Íleon/cirugía , Obstrucción Intestinal/etiología , Leiomiomatosis/complicaciones , Persona de Mediana Edad , Neoplasias Peritoneales/complicaciones , Neoplasias del Recto/complicaciones , Neoplasias del Recto/cirugía , Enfermedades del Sigmoide/etiologíaRESUMEN
We have evaluated whether i.p. murine ovarian tumors could be treated with an IL-2 plasmid DNA complexed with the cationic lipid, (+/-)-N-(2-hydroxyethyl)-N,N-dimethyl-2, 3-bis(tetradecyloxy)-1-propanaminium bromide/dioleoylphosphatidylethanolamine (DMRIE/DOPE). Reporter gene studies were initially conducted in which mice bearing i.p. murine ovarian teratocarcinoma (MOT) were injected i.p. with reporter gene plasmid DNA (pDNA):DMRIE/DOPE. Histochemical analyses revealed that transfection occurred primarily in the tumor cells of the ascites, with only a minority of other ascitic cells or surrounding tissues transfected. IL-2 levels in the MOT ascites were determined after i. p. injection of either IL-2 pDNA:DMRIE/DOPE or recombinant IL-2 protein. IL-2 was detected in tumor ascites for up to 10 days after a single i.p. injection of IL-2 pDNA:DMRIE/DOPE, but was undetectable 24 h after a single i.p. injection of IL-2 protein. In an antitumor efficacy study, MOT tumor-bearing mice injected i.p. with IL-2 pDNA:DMRIE/DOPE on days 5, 8, and 11 after tumor cell implant had a significant inhibition of tumor ascites (p = 0.001) as well as a significant increase in survival (p = 0.008). A cytokine profile of the MOT tumor ascites revealed that mice treated with IL-2 pDNA:DMRIE/DOPE had an IL-2-specific increase in the levels of IFN-gamma and GM-CSF. Taken together, these findings indicate that i. p. treatment of ovarian tumors with IL-2 pDNA:DMRIE/DOPE can lead to an increase in local IL-2 levels, a change in the cytokine profile of the tumor ascites, and a significant antitumor effect.
Asunto(s)
Ascitis/prevención & control , Citocinas/biosíntesis , Inhibidores de Crecimiento/uso terapéutico , Interleucina-2/genética , Interleucina-2/uso terapéutico , Neoplasias Ováricas/terapia , Plásmidos/uso terapéutico , Teratocarcinoma/terapia , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/inmunología , Antineoplásicos/uso terapéutico , Ascitis/metabolismo , Ascitis/patología , ADN Bacteriano/administración & dosificación , ADN Bacteriano/genética , Relación Dosis-Respuesta Inmunológica , Femenino , Inhibidores de Crecimiento/administración & dosificación , Inhibidores de Crecimiento/genética , Inhibidores de Crecimiento/inmunología , Inyecciones Intraperitoneales , Interleucina-2/administración & dosificación , Lípidos/administración & dosificación , Ratones , Ratones Endogámicos C3H , Ratones Desnudos , Neoplasias Ováricas/química , Neoplasias Ováricas/genética , Neoplasias Ováricas/inmunología , Fosfatidiletanolaminas/administración & dosificación , Fosfatidiletanolaminas/genética , Plásmidos/administración & dosificación , Plásmidos/inmunología , Compuestos de Amonio Cuaternario/administración & dosificación , Teratocarcinoma/química , Teratocarcinoma/genética , Teratocarcinoma/inmunologíaRESUMEN
The immunosuppressive protein transforming growth factor beta (TGF-beta) inhibits the activation of various immune effector cells including cytotoxic T lymphocytes and may therefore inhibit the efficacy of immunostimulatory interleukin-2 (IL-2) gene therapy. In this study, we investigated the effect of TGF-beta downregulation on IL-2 gene therapy in the intraperitoneal model of murine ovarian teratoma (MOT). MOT cells, like many human ovarian carcinomas, were found to produce TGF-beta. Production of TGF-beta by MOT cells was suppressed using a TGF-beta antisense plasmid vector (pCEP4/TGF-beta antisense). Subcutaneous immunization of C3H mice with a mixture of IL-2 gene-transduced fibroblasts and TGF-beta antisense-modified MOT cells induced significantly better protection against a subsequent intraperitoneal tumor challenge compared with immunization with unmodified MOT cells alone [11/16 (69%) vs 4/21 (19%) tumor-free animals, P < 0.01]. Immunization with either a mixture of IL-2 gene modified fibroblasts and unmodified MOT cells [2/12 (17%) tumor-free animals] or TGF-beta antisense-modified MOT cells alone (0/13 tumor free animals) failed to induce significant protection compared with immunization with unmodified MOT cells. These data show that combined TGF-beta antisense and IL-2 gene therapy is required to generate effective antitumor responses in the MOT model. Our findings suggest that tumor cell expression of immunosuppressive factors may inhibit cytokine immunogene therapy and may have potential implications for the development of future clinical immunogene therapy protocols.
Asunto(s)
Elementos sin Sentido (Genética)/uso terapéutico , Vacunas contra el Cáncer/uso terapéutico , Terapia Genética , Interleucina-2/genética , Neoplasias Ováricas/terapia , Teratoma/terapia , Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Células 3T3 , Animales , Regulación hacia Abajo , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3HRESUMEN
OBJECT: To study the combined potential of wild-type p53 gene transfer and administration of cisplatin for the treatment of glioblastoma multiforme, the authors used the 9L rat glioblastoma cell line, which expresses a mutant p53. METHODS: Stable expression of wild-type p53 in 9L cells was achieved by transfection of the cells with a wild-type p53-expressing plasmid (pCEP4p53). The resultant cell line, 9LpCEP4p53, was found to be more sensitive to cisplatin treatment in vitro than control (9LpCEP4) cells. The in vitro growth rates of control cells and wild-type p53-modified cells were similar in the absence of cisplatin. Fischer 344 rats were implanted intracerebrally with 9LpCEP4p53 cells and intraperitoneally administered 4 mg/kg cisplatin weekly for 7 weeks. These animals survived significantly longer than animals that were implanted with 9LpCEP4p53 cells but were given no cisplatin treatment. In contrast, concurrent cisplatin treatment provided no benefit for animals implanted with 9LpCEP4 cells. Tumors that developed in animals that had been implanted with 9LpCEP4p53 cells and treated with cisplatin had lost expression of wild-type p53, indicating a correlation between expression of wild-type p53 and cisplatin sensitivity in vivo. CONCLUSIONS: The findings of this study suggest that p53-based gene therapy in combination with cisplatin-based chemotherapy may be superior to single-modality treatment in dealing with glioblastoma multiforme.
Asunto(s)
Antineoplásicos/uso terapéutico , Cisplatino/uso terapéutico , Resistencia a Antineoplásicos/genética , Genes p53/genética , Glioblastoma/genética , Mutación/genética , Animales , Antineoplásicos/administración & dosificación , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/genética , División Celular/efectos de los fármacos , Supervivencia Celular , Cisplatino/administración & dosificación , Terapia Combinada , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Técnicas de Transferencia de Gen , Terapia Genética , Glioblastoma/tratamiento farmacológico , Herpesvirus Humano 4/genética , Inyecciones Intraperitoneales , Trasplante de Neoplasias , Plásmidos , Ratas , Ratas Endogámicas F344 , Análisis de Regresión , Tasa de Supervivencia , Transfección , Células Tumorales CultivadasRESUMEN
Several investigators have employed interleukin-2 (IL-2) gene transfer to enhance the immunogenicity of tumor cell vaccines. We describe in this report the construction and characterization of retroviral vectors for IL-2 gene therapy. Human IL-2 cDNA with a chimeric rat preproinsulin/IL-2 DNA leader sequence was subcloned into the pLXSN (long terminal repeat promoter) and pLNCX (cytomegalovirus [CMV] promoter) vectors to generate the plasmids pLXSN-iIL2 and pLNCX-iIL2, respectively. Human IL-2 cDNA with a chimeric human tissue factor/IL-2 DNA leader sequence was utilized to construct the vector pLXSN-tIL2. The levels of IL-2 secreted by transduced tumor cells and fibroblasts were evaluated by enzyme-linked immunosorbent assay (ELISA) of culture supernatants and compared with those of normal peripheral blood mononuclear cells (PBMC) activated in vitro with calcium ionophore and phorbol 12-myristate 13-acetate. The highest levels of IL-2 secreted by transduced tumor cells (760 units/10(6) cells/24 h), adult fibroblasts (625 units/10(6) cells/24 h), and embryonic fibroblasts (3,975 units/10(6) cells/24 h) were 150- to 1,000-fold higher than than secreted by the activated PBMC (4 units/10(6) cells/24 h). Similar levels of IL-2 were expressed by human fibroblasts transduced with pLXSN vectors employing the preproinsulin (pLXSN-iIL2) or tissue factor (pLXSN-tIL2) leader sequences (range in IL-2 units/10(6) cells/24 h pLXSN-iIL2 = 375-625 vs. pLXSN-tIL2 = 90-440). Because IL-2-transduced cells for clinical applications are generally irradiated to prevent cellular proliferation, we evaluated the effects of radiation on IL-2 production. Radiation doses between 1,500 and 10,000 cGy resulted in gradual decreases in IL-2 secretion by transduced cells. The range of the decrease in IL-2 secretion was 7-11% by day 7, 0-29% by day 14, and 25-50% by day 35. For clinical applications, stable production of the vector in high concentrations is an important consideration. The retroviral vector pLXSN-tIL2 produced the highest viral titer and was chosen for further characterization. Southern blot analysis of SacI-digested genomic DNA from the LXSN-tIL2 producer cell line and SacI-digested pLXSN-tIL2 plasmid DNA revealed the expected 3.2-kbp fragment, suggesting the absence of transgene rearrangement and the suitability of this vector as a candidate for clinical applications.
Asunto(s)
Terapia Genética , Vectores Genéticos , Interleucina-2/genética , Retroviridae/genética , Células 3T3 , Animales , Secuencia de Bases , Línea Celular , Expresión Génica , Técnicas de Transferencia de Gen , Humanos , Insulina , Linfocitos/metabolismo , Ratones , Plásmidos , Reacción en Cadena de la Polimerasa , Proinsulina/genética , Precursores de Proteínas/genética , ARN Mensajero/análisis , Ratas , Tromboplastina/genéticaRESUMEN
In the last decade, advances in molecular biology have lead to the development of techniques that permit the manipulation of mammalian cell DNA for diagnostic and therapeutic purposes. Gene therapy has subsequently evolved as a treatment option in patients with malignancies. In this article, we have summarized current strategies in gene therapy for ovarian cancer.
RESUMEN
We previously demonstrated the efficacy of a genetically engineered vaccine composed of syngeneic tumor cells mixed with syngeneic, IL-2 gene transduced fibroblasts in the CT-26 model of murine colorectal carcinoma. In this report, we describe a more practical approach to fibroblast mediated IL-2 gene therapy that employs syngeneic tumor cells mixed with allogeneic, IL-2 gene transduced fibroblasts. BALB/c mice were challenged with an injection of CT-26, 14 days following immunization with IL-2 modified syngeneic BALB/c 3T3 (H-2(d)) or allogeneic C3H 3T3 (H-2(k)) fibroblasts mixed with irradiated CT-26. Both syngeneic and allogeneic IL-2 modified fibroblasts provided significantly better protection compared to animals treated with control fibroblasts (syngeneic IL-2 fibroblasts 32/40-80% vs. control 15/45-33%, p<0.01; allogeneic IL-2 fibroblasts 25/37-68% vs. control 15/45-3345, p<0.01). There was no statistically significant difference between the groups immunized with syngeneic or allogeneic IL-2 modified fibroblasts. These findings support the evaluation of allogeneic IL-2 modified fibroblasts as a practical form of cytokine gene therapy for cancer.
RESUMEN
Like human gliomas, the rat 9L gliosarcoma secretes the immunosuppressive transforming growth factor beta (TGF-beta). Using the 9L model, we tested our hypothesis that genetic modification of glioma cells to block TGF-beta expression may enhance their immunogenicity and make them more suitable for active tumor immunotherapy. Subcutaneous immunizations of tumor-bearing animals with 9L cells genetically modified to inhibit TGF-beta expression with an antisense plasmid vector resulted in a significantly higher number of animals surviving for 12 weeks (11/11, 100%) compared to immunizations with control vector-modified 9L cells (2/15, 13%) or 9L cells transduced with an interleukin 2 retroviral vector (3/10, 30%) (P < 0.001 for both comparisons). Histologic evaluation of implantation sites 12 weeks after treatment revealed no evidence of residual tumor. In vitro tumor cytotoxicity assays with lymph node effector cells revealed a 3- to 4-fold increase in lytic activity for the animals immunized with TGF-beta antisense-modified tumor cells compared to immunizations with control vector or interleukin 2 gene-modified tumor cells. These results indicate that inhibition of TGF-beta expression significantly enhances tumor-cell immunogenicity and supports future clinical evaluation of TGF-beta antisense gene therapy for TGF-beta-expressing tumors.
Asunto(s)
Neoplasias Encefálicas/terapia , ADN sin Sentido/uso terapéutico , Terapia Genética , Gliosarcoma/terapia , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Crecimiento Transformador beta/genética , Animales , Neoplasias Encefálicas/patología , Citotoxicidad Inmunológica , ADN sin Sentido/administración & dosificación , Expresión Génica/efectos de los fármacos , Terapia Genética/métodos , Vectores Genéticos , Gliosarcoma/patología , Humanos , Inmunoterapia/métodos , Interleucina-2/biosíntesis , Linfocitos/inmunología , Plásmidos , Ratas , Ratas Endogámicas F344 , Retroviridae , Factores de TiempoRESUMEN
We compared the efficacy of gene therapy mediated by interleukin-2 (IL-2) gene-modified tumor cells to gene therapy mediated by IL-2 transduced fibroblasts in the CT-26 model of murine colorectal carcinoma. We transduced CT-26 tumor cells and BALB/c 3T3 fibroblasts with three different retroviral vectors using three different promoters for the human IL-2 gene: DC/TKIL-2 (thymidine kinase promoter), LXSN-iIL2 (long terminal repeat promoter), and LNCX-iIL2 (cytomegalovirus promoter). These transductions resulted in CT-26 and 3T3 subclones that secreted different amounts of IL-2. Immunization of animals with either CT-26/IL-2 cells or with fibroblast/IL-2 cells mixed with CT-26 induced similar levels of immunity that protected 62-82% of animals against a subsequent tumor challenge with parental CT-26. However, mice developed tumors at the site of inoculation in 46% of the animals immunized with CT-26/IL-2 cells. In a separate experiment, CT-26/IL-2 cells were exposed to 6,000 cGy of gamma irradiation to prevent tumor growth at the site of inoculation. Although the CT-26/IL-2 cells continued to secrete IL-2 after irradiation, they were no longer effective at inducing antitumor immunity. In contrast, both irradiated and nonirradiated fibroblast/IL-2 cells, mixed with irradiated CT-26, were equally effective at inducing antitumor immunity. These data suggest that in the CT-26 model, fibroblast-mediated IL-2 gene therapy has advantages for the induction of antitumor immunity and abrogation of tumorigenic potential at the site of inoculation compared with tumor cell-mediated IL-2 gene therapy.
Asunto(s)
Células 3T3/efectos de los fármacos , Neoplasias Colorrectales/terapia , Terapia Genética , Interleucina-2/genética , Transfección , Células 3T3/efectos de la radiación , Animales , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/inmunología , Modelos Animales de Enfermedad , Humanos , Interleucina-2/farmacología , Interleucina-2/efectos de la radiación , Ratones , Ratones Endogámicos BALB C , Células Tumorales CultivadasRESUMEN
A patient with glioblastoma multiforme (GBM) who had failed conventional therapy was treated with IL-2 gene therapy. The patient received 10 subcutaneous immunizations with autologous tumor cells and fibroblasts genetically modified to secrete IL-2 by retroviral gene transfer. An antitumor immune response mediated in part by CD8+ cytotoxic T cells was demonstrated with the patient's peripheral blood mononuclear cells. A magnetic resonance imaging (MRI) scan performed 4 weeks after the highest treatment dose revealed marked tumor necrosis. These results support the evaluation of this form of IL-2 gene therapy in additional patients with glioblastoma.
Asunto(s)
Neoplasias Encefálicas/terapia , Fibroblastos/metabolismo , Fibroblastos/trasplante , Terapia Genética , Glioblastoma/terapia , Interleucina-2/uso terapéutico , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/trasplante , Proteínas Recombinantes de Fusión/uso terapéutico , Terapia Recuperativa , Lóbulo Temporal , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias Encefálicas/inmunología , Neoplasias Encefálicas/patología , Células Cultivadas/trasplante , Terapia Combinada , Resultado Fatal , Femenino , Vectores Genéticos , Glioblastoma/inmunología , Glioblastoma/patología , Humanos , Inyecciones Subcutáneas , Interleucina-2/genética , Interleucina-2/metabolismo , Isotretinoína/uso terapéutico , Lomustina/administración & dosificación , Imagen por Resonancia Magnética , Persona de Mediana Edad , Necrosis , Recurrencia Local de Neoplasia , Procarbazina/administración & dosificación , Radioinmunoterapia , Radiocirugia , Proteínas Recombinantes de Fusión/metabolismo , Retroviridae/genética , Linfocitos T Citotóxicos/inmunología , Tamoxifeno/uso terapéutico , Transfección , Células Tumorales Cultivadas/trasplante , Vincristina/administración & dosificaciónRESUMEN
We have established and characterized a new glioblastoma cell line, termed GT9, from a biopsy sample of a female adult patient with glioblastoma multiforme. The line has now undergone over 60 passages and has been successfully cultured after cryopreservation. Immunofluorescence analyses with a panel of monoclonal antibodies were positive for glial fibrillary acidic protein and vimentin, and negative for neurofilament, galactocerebroside, and fibronectin, a pattern typical of glial cells. Based on a tetraploid, the composite karyotype of GT9 cells included the loss of chromosome 10, gain of chromosome 7, and the presence of double minute chromosomes, three of the most common karyotypic abnormalities in glioblastoma. Sequence analysis of p53 cDNA revealed a homozygous double mutation at codon 249 (commonly mutated in aflatoxin-associated hepatocellular carcinoma) and codon 250. Moreover, there was a complete absence of wild-type p53. However, unlike the majority of human glioblastomas previously described, the expression of platelet-derived growth factor-B (PDGF-B), a potent mitogenic autocrine factor, was low in GT9 cells. The expression and phosphorylation of c-Jun and Jun-B, downstream mediators of the PDGF pathway, were also low. Thus, deregulation of the PDGF pathway does not appear to be involved in the pathogenesis of the GT9 glioblastoma. Conversely, Jun-D, a negative regulator of cell growth, was also low. In addition, Phosphorylated Egr-1, a recently reported suppressor of PDGF-B/v-sis-transformed cells, was also low, suggesting that the lack of activation of the PDGF pathway was not due to these suppressive mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Genes p53 , Glioblastoma/genética , Proteínas Inmediatas-Precoces , Mutación , Factor de Crecimiento Derivado de Plaquetas/metabolismo , Proteína p53 Supresora de Tumor/genética , Secuencia de Aminoácidos , Anticuerpos Monoclonales , Aberraciones Cromosómicas , Cromosomas Humanos Par 10 , Cromosomas Humanos Par 7 , Proteínas de Unión al ADN/metabolismo , Proteína 1 de la Respuesta de Crecimiento Precoz , Femenino , Técnica del Anticuerpo Fluorescente , Proteína Ácida Fibrilar de la Glía/análisis , Glioblastoma/patología , Humanos , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas Proto-Oncogénicas c-jun/metabolismo , Factores de Transcripción/metabolismo , Células Tumorales Cultivadas , Vimentina/análisisAsunto(s)
Ensayos Clínicos Fase I como Asunto , Neoplasias del Colon/terapia , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Interleucina-2/genética , Trasplante de Células , Protocolos Clínicos , Fibroblastos , Técnicas de Transferencia de Gen/efectos adversos , Humanos , Consentimiento Informado , Interleucina-2/uso terapéuticoRESUMEN
In 11 rats, the microcirculation of the repeatedly ischaemic (stunned) left ventricular myocardium was studied using in vivo fluorescence microscopy. Stunning was provoked by six subsequent 10 min ligations of the left anterior descending coronary artery, each of them followed by a 20 min reperfusion period. In the stunned myocardium showing hypokinetic wall motion, myocardial blood flow dropped by 55%; in this region, leukocytes often appeared in slow-flow capillaries plugging capillary branches. Closely linking to leukocyte adherence, a rise of microvascular permeability was documented by extravascular clouds of fluorescent dextran. After nifedipine treatment, in ischaemic regions marked dilatation of larger A1 and A2 arterioles was noted, in addition to the ischaemia-induced dilatation of smaller A3 and A4 arterioles. Furthermore nifedipine and nisoldipine reduced the number of adherent leukocytes in post-capillary venules and capillaries of the repeatedly ischaemic myocardium. In 12 patients with coronary one-vessel disease and without previous transmural myocardial infarction, elective coronary angioplasty (PTCA) was performed (balloon inflation for 2 min). After elective PTCA of the LAD, a significant rise in the proportion of activated neutrophils was noted. After elective 2 min PTCA of the LAD, coronary sinus blood samples showed a marked rise of FMLC stimulated superoxide anion production, whereas passive deformability decreased considerably. Furthermore, an increase in chemotactic activity in coronary sinus blood samples was observed.
Asunto(s)
Circulación Coronaria/fisiología , Aturdimiento Miocárdico/fisiopatología , Neutrófilos/fisiología , Angioplastia Coronaria con Balón , Animales , Velocidad del Flujo Sanguíneo , Permeabilidad Capilar , Adhesión Celular , Circulación Coronaria/efectos de los fármacos , Circulación Coronaria/inmunología , Vasos Coronarios/inmunología , Vasos Coronarios/fisiopatología , Humanos , Leucocitos/fisiología , Microcirculación/efectos de los fármacos , Microcirculación/inmunología , Microcirculación/fisiopatología , Aturdimiento Miocárdico/inmunología , Nifedipino/farmacología , RatasRESUMEN
The 1F7 idiotype previously defined (J Immunol 147:933;1991 and Eur J Immunol 22:1749;1992) is expressed on antibodies reactive to different proteins of HIV (gp41, p24, gp120). Since serum levels of 1F7 (IgM or IgG) are significantly higher in patients with HIV lymphoma as opposed to HIV-infected individuals, normal controls and non-HIV lymphoma patients, we hypothesized the B-cell neoplasms were the source of the idiotype. However, immunohistochemistry on cryostat sections revealed no 1F7 idiotype signal on neoplastic B-cells nor tumor infiltrative T-cells (n = 8). Interestingly, reactive lymphocytosis adjacent to tumor masses and reactive follicular hyperplastic controls (n = 5) exhibited significant 1F7 reactivity. The reactivity appeared in paracortical and perifollicular lymphoid regions, predominantly regions of B, T and antigen presenting cells in lymph nodes or tonsils. A survey of electrophoretically defined paraproteins with anti-HIV specificities derived from HIV-infected patients showed no Western blot reactivity with the 1F7 anti-idiotypic antibody. Therefore, the idiotype does not appear to be a direct product of B-cell neoplasia or abnormal B-cell proliferation, but is produced by B cell clones responding to HIV infection. This high level of serum 1F7 reactivity could be an important clue in the pathogenesis of HIV lymphomas and confers a highly predictive serological test for HIV lymphoma.