RESUMEN
The selective targeting of the tumor-associated death-inducing receptors DR4 and DR5 with agonistic mAbs has demonstrated preclinical and clinical antitumor activity. However, the cellular and molecular mechanisms contributing to this efficacy remain poorly understood. In this study, using the first described C57BL/6 (B6) TRAIL-sensitive experimental tumor models, we have characterized the innate and adaptive immune components involved in the primary rejection phase of an anti-mouse DR5 (mDR5) mAb, MD5-1 in established MC38 colon adenocarcinomas. FcR mediated cross-linking of MD5-1 significantly inhibited the growth of MC38 colon adenocarcinomas through the induction of TRAIL-R-dependent tumor cell apoptosis. The loss of host DR5, TRAIL, perforin, FasL, or TNF did not compromise anti-DR5 therapy in vivo. By contrast, anti-DR5 therapy was completely abrogated in mice deficient of B cells or CD11c(+) dendritic cells (DCs), providing the first direct evidence that these cells play a critical role. Importantly, the requirement for an intact B cell compartment for optimal anti-DR5 antitumor efficacy was also observed in established AT-3 mammary tumors. Interestingly, MD5-1-mediated apoptosis as measured by early TUNEL activity was completely lost in B cell-deficient microMT mice, but intact in mice deficient in CD11c(+) DCs. Overall, these data show that Ab-mediated targeting of DR5 triggers tumor cell apoptosis in established tumors in a B cell-dependent manner and that CD11c(+) DCs make a critical downstream contribution to anti-DR5 antitumor activity.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Citotoxicidad Celular Dependiente de Anticuerpos/inmunología , Subgrupos de Linfocitos B/inmunología , Antígeno CD11c/fisiología , Células Dendríticas/inmunología , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/inmunología , Adenocarcinoma/inmunología , Adenocarcinoma/patología , Adenocarcinoma/terapia , Animales , Anticuerpos Monoclonales/toxicidad , Subgrupos de Linfocitos B/metabolismo , Antígeno CD11c/biosíntesis , Línea Celular , Línea Celular Tumoral , Neoplasias del Colon/inmunología , Neoplasias del Colon/patología , Neoplasias del Colon/terapia , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo/inmunología , Humanos , Linfoma de Células B/inmunología , Linfoma de Células B/terapia , Neoplasias Mamarias Experimentales/inmunología , Neoplasias Mamarias Experimentales/terapia , Mastocitoma/inmunología , Mastocitoma/terapia , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/deficiencia , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/genéticaRESUMEN
Therapeutic vaccination regimens inducing clinically effective tumor-specific CD8+ T lymphocyte (CTL) responses are an unmet medical need. We engineer two distantly related arenaviruses, Pichinde virus and lymphocytic choriomeningitis virus, for therapeutic cancer vaccination. In mice, life-replicating vector formats of these two viruses delivering a self-antigen in a heterologous prime-boost regimen induce tumor-specific CTL responses up to 50% of the circulating CD8 T cell pool. This CTL attack eliminates established solid tumors in a significant proportion of animals, accompanied by protection against tumor rechallenge. The magnitude of CTL responses is alarmin driven and requires combining two genealogically distantly related arenaviruses. Vector-neutralizing antibodies do not inhibit booster immunizations by the same vector or by closely related vectors. Rather, CTL immunodominance hierarchies favor vector backbone-targeted responses at the expense of self-reactive CTLs. These findings establish an arenavirus-based immunotherapy regimen that allows reshuffling of immunodominance hierarchies and breaking self-directed tolerance for efficient tumor control.
Asunto(s)
Vacunas contra el Cáncer/administración & dosificación , Inmunoterapia/métodos , Virus de la Coriomeningitis Linfocítica/inmunología , Mastocitoma/terapia , Virus Pichinde/inmunología , Linfocitos T Citotóxicos/inmunología , Alarminas/genética , Alarminas/inmunología , Animales , Anticuerpos Neutralizantes/farmacología , Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/inmunología , Femenino , Expresión Génica , Ingeniería Genética/métodos , Vectores Genéticos/clasificación , Vectores Genéticos/inmunología , Cobayas , Inmunización Secundaria , Virus de la Coriomeningitis Linfocítica/clasificación , Virus de la Coriomeningitis Linfocítica/genética , Mastocitoma/genética , Mastocitoma/inmunología , Mastocitoma/mortalidad , Ratones , Ratones Endogámicos C57BL , Filogenia , Virus Pichinde/clasificación , Virus Pichinde/genética , Autotolerancia , Análisis de Supervivencia , Vacunación/métodosRESUMEN
Mast cell tumor (MCT) or mastocytoma is one of the most frequent malignant cutaneous tumors in the dog, and the second most frequent in the cat. Several mast cell tumor therapeutic approaches have been proposed in the past years for dogs and cats, resulting in very distinct outcomes. The current comprehensive literature review presents a critical approach to the scientific information published about the MCTs treatments and the subsequent prognosis and survival times, in dogs and in cats diagnosed with MCTs. A systematic review of the literature following the Cochrane principles and methodology was performed. The authors resorted to MEDLINE, Scopus, Google Scholar and Web of Science databases to select the 133 publications with evidence-based treatments for MCTs in companion animals. Results of the review suggest that the recommended treatment, prognosis and survival times for dogs and cats with MCTs depends at all times on the clinical staging, histological grade and location of the tumor.
Asunto(s)
Enfermedades de los Gatos/terapia , Enfermedades de los Perros/terapia , Mastocitoma/veterinaria , Neoplasias Cutáneas/veterinaria , Animales , Gatos , Perros , Mastocitoma/terapia , Neoplasias Cutáneas/terapia , Resultado del TratamientoRESUMEN
Tumor-associated macrophages (TAMs) are frequently found in glioblastomas and a high degree of macrophage infiltration is associated with a poor prognosis for glioblastoma patients. However, it is unclear whether TAMs in glioblastomas promote tumor growth. In this study, we found that folate receptor beta (FR beta) was expressed on macrophages in human glioblastomas and a rat C6 glioma implanted subcutaneously in nude mice. To target FR beta-expressing TAMs, we produced a recombinant immunotoxin consisting of immunoglobulin heavy and light chain Fv portions of an anti-mouse FR beta monoclonal antibody and Pseudomonas exotoxin A. Injection of the immunotoxin into C6 glioma xenografts in nude mice significantly depleted TAMs and reduced tumor growth. The immunotoxin targeting FR beta-expressing macrophages will provide a therapeutic tool for human glioblastomas.
Asunto(s)
ADP Ribosa Transferasas/uso terapéutico , Toxinas Bacterianas/uso terapéutico , Proteínas Portadoras/inmunología , Exotoxinas/uso terapéutico , Glioblastoma/terapia , Inmunotoxinas/uso terapéutico , Macrófagos Peritoneales/inmunología , Receptores de Superficie Celular/inmunología , Proteínas Recombinantes de Fusión/uso terapéutico , Factores de Virulencia/uso terapéutico , Animales , Anticuerpos Monoclonales/inmunología , Citotoxicidad Celular Dependiente de Anticuerpos , Femenino , Receptores de Folato Anclados a GPI , Glioblastoma/inmunología , Glioblastoma/patología , Técnicas para Inmunoenzimas , Región Variable de Inmunoglobulina/inmunología , Mastocitoma/inmunología , Mastocitoma/terapia , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Mieloma Múltiple/inmunología , Mieloma Múltiple/terapia , Óxido Nítrico/metabolismo , Ratas , Tasa de Supervivencia , Factor A de Crecimiento Endotelial Vascular/metabolismo , Ensayos Antitumor por Modelo de Xenoinjerto , Exotoxina A de Pseudomonas aeruginosaRESUMEN
A strong CTL response is dependent upon a high level of expression of specific class I major histocompatibility complex (MHC)/peptide complexes at the cell surface. An epitope-linked beta2-microglobulin (beta2m) molecule could provide a simple and more efficient means to enhance the formation of defined MHC/peptide complexes. However, the ability of an epitope-linked beta2m molecule to elicit primary CTL responses in vivo is still unknown. In this study, we modified the P1A tumor cell vaccine by addition of the tumor-associated epitope (TAE)-linked beta2m molecule and co-stimulatory molecule CD80 to improve the efficiency in the application of the vaccine. A eukaryotic co-expression vector consisting of the P1A35-43-linked beta2m molecule and the murine CD80 gene was constructed. P815 cell lines stably expressing P1A35-43-linked beta2m molecule and/or CD80 were established after transfection, by selection under G418. Administration of these inactivated tumor cell vaccines allowed the TAE-specific CD8+ T cell responses to be examined in vivo. Our results indicate that immunization with P815 cells expressing both the P1A35-43-linked beta2m molecule and the murine CD80 gene elicited a significantly stronger antitumor immune response than the single-modified tumor cell vaccines (expressing either P1A35-43-linked beta2m or CD80 alone). These findings support the feasibility and effectiveness of developing a dual-modified tumor cell vaccine consisting of the epitope-linked beta2m molecule and a co-stimulatory molecule.
Asunto(s)
Antígenos de Neoplasias/inmunología , Antígeno B7-1/inmunología , Vacunas contra el Cáncer/uso terapéutico , Mastocitoma/inmunología , Mastocitoma/terapia , Microglobulina beta-2/inmunología , Animales , Antígenos de Neoplasias/genética , Antígeno B7-1/genética , Linfocitos T CD8-positivos/inmunología , Estudios de Factibilidad , Femenino , Inmunización , Interferón gamma , Mastocitoma/genética , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Bazo/inmunología , Linfocitos T Citotóxicos/inmunología , Microglobulina beta-2/genéticaRESUMEN
UNLABELLED: The aim of this study was to evaluate the effectiveness of local treatment electrochemotherapy (ECT) with cisplatin and to compare it with effectiveness of surgery for treatment of mast cell tumours (MCT) in dogs. MATERIALS AND METHODS: In the present retrospective study, 25 dogs of different breeds with MCT were divided into two treatment groups: surgery (16 dogs with 16 tumours) and those whose owners refused surgery being included into the ECT group (9 dogs with 12 tumours). Response rate and duration of response to the treatment were evaluated and comparison between groups was made. RESULTS: The clinical stages of the tumours were stage I in 4 (45%) and stage III in 5 (55%) dogs treated by ECT; 12 (75%) dogs treated by surgery were stage I and 4 (25%) dogs were in clinical stage III. The median size of the tumours was 5.2 cm3 and 2.9 cm3 of tumours treated by surgery and ECT, respectively. ECT resulted in as comparable antitumor effectiveness as surgical treatment. However, the estimated median duration of response in dogs treated with complete surgical excision was 31.5 months, while it was not reached for the ECT group at the time of writing. CLINICAL SIGNIFICANCE: ECT is an easy, effective and safe local treatment of MCT. It can be an alternative treatment to surgery, specifically for smaller nodules in which a complete response with long duration can be obtained after only one treatment session, or when the nodule is unresectable because of the location.
Asunto(s)
Antineoplásicos/administración & dosificación , Cisplatino/administración & dosificación , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/cirugía , Electroquimioterapia/veterinaria , Mastocitoma/veterinaria , Neoplasias Cutáneas/veterinaria , Animales , Terapia Combinada/veterinaria , Perros , Electroporación/veterinaria , Femenino , Masculino , Mastocitoma/terapia , Estadificación de Neoplasias/veterinaria , Inducción de Remisión , Estudios Retrospectivos , Neoplasias Cutáneas/terapiaRESUMEN
BACKGROUND: Many viruses have been engineered and evaluated for their potential as therapeutic agents in the treatment of malignant neoplasm, including malignant melanoma. OBJECTIVE: In this study, we investigated the efficacy of HF10, an attenuated, replication-competent HSV, in immunocompetent animal models with malignant melanoma. METHODS: For in vitro study, viral cytotoxicity assays and replication assays were performed both in human and mouse melanoma cells. For the study in vivo, intraperitoneally disseminated or subcutaneous melanoma models were prepared in DBA/2 mice using clone M3 cells, then HF10 was inoculated intraperitoneally or intratumorally. Therapeutic efficacy of HF10 was assessed by survival, tumor growth, and histopathological analysis. RESULTS: HF10 infection produced cytolytic effects in melanoma cells at various multiplicities of infection (MOI). In the intraperitoneal melanoma model, all mice survived when given intraperitoneal injections of HF10 compared with 100% fatality in the control mice. In the subcutaneous tumor model, intratumoral inoculation of HF10 significantly reduced tumor growth. Histology and immunohistochemistry showed tumor lysis and inflammatory cell infiltration after intratumoral HF10 inoculation. Viral antigen was retained at the inoculation site until 7 days post-infection. HF10-treated intraperitoneal tumor mice were also protected against tumor rechallenge. HF10 also affected the non-inoculated contralateral tumor when injected into the ipsilateral tumor of mice, suggesting that HF10 can induce systemic antitumor immune responses in mice. CONCLUSION: Oncolytic viral therapy using HF10 was effective in melanoma mouse models, and intratumoral injection of HF10 induced systemic antitumor responses. These results suggest that HF10 is a promising agent for the treatment of advanced melanoma.
Asunto(s)
Herpesvirus Humano 1/genética , Melanoma/terapia , Viroterapia Oncolítica/métodos , Neoplasias Cutáneas/terapia , Animales , Línea Celular Tumoral , Chlorocebus aethiops , Modelos Animales de Enfermedad , Femenino , Herpesvirus Humano 1/crecimiento & desarrollo , Herpesvirus Humano 1/inmunología , Humanos , Inmunocompetencia , Inyecciones Intraperitoneales , Inyecciones Subcutáneas , Mastocitoma/inmunología , Mastocitoma/patología , Mastocitoma/terapia , Melanoma/inmunología , Melanoma/patología , Ratones , Ratones Endogámicos DBA , Necrosis , Trasplante de Neoplasias/métodos , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patología , Tasa de Supervivencia , Células VeroRESUMEN
DNA vaccination against cancer has become a promising strategy for inducing a specific and long-lasting antitumor immunity. However, DNA vaccines fail to generate potent immune responses when used as a single therapy. To enhance their activity into the tumor, a DNA vaccine against murine P815 mastocytoma was combined with antibodies directed against the immune checkpoints CTLA4 and PD1. The combination of these two strategies delayed tumor growth and enhanced specific antitumor immune cell infiltration in comparison to the corresponding single therapies. The combination also promoted IFNg, IL12 and granzyme B production in the tumor microenvironment and decreased the formation of liver metastasis in a very early phase of tumor development, enabling 90% survival. These results underline the complementarity of DNA vaccination and immune checkpoint blockers in inducing a potent immune response, by exploiting the generation of antigen-specific T cells by the vaccine and the ability of immune checkpoint blockers to enhance T cell activity and infiltration in the tumor. These findings suggest how and why a rational combination therapy can overcome the limits of DNA vaccination but could also allow responses to immune checkpoint blockers in a larger proportion of subjects.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Vacunas contra el Cáncer/uso terapéutico , Mastocitoma/terapia , Receptor de Muerte Celular Programada 1/inmunología , Vacunas de ADN/uso terapéutico , Animales , Antígeno CTLA-4/inmunología , Vacunas contra el Cáncer/inmunología , Inmunoterapia/métodos , Mastocitoma/patología , Ratones , Metástasis de la Neoplasia/prevención & control , Tasa de Supervivencia , Resultado del Tratamiento , Microambiente Tumoral , Vacunas de ADN/inmunologíaRESUMEN
Cancer immunotherapy has attracted much attention in recent years because of the ability of immune system to identify tumor cells and limit their growth. Icariin (ICA) is a natural flavonoid glucoside isolated from Epimedium plants and has shown a variety of pharmacological activities such as anti-inflammatory effects, immunological regulation and anticancer potency. Furthermore, it has immunoadjuvant effects on enhancing Th1-immune response, suggesting that ICA may serve as an adjuvant for cancer immunotherapy. In this study, we used P815 mouse mastocytoma tumor model and immunized them with P815AB peptide and/or ICA. Our results demonstrated that ICA could increase the cytotoxic T lymphocytes (CTL) response for P815AB peptide on the tumor-bearing DBA/2J mice. In addition, the percentage of CD4+CD8+/CD3+CD69+/CD69+NKG2D+ positive cells in splenocytes of the tumor-bearing mice all significantly increased after combined immunization with ICA and P815AB peptide. This illustrated that ICA could enhance the immunogenicity of P815AB and improve the ability of T cells and CTLs in recognizing the tumor cells. Moreover, ICA improved the function of peritoneal macrophages with effects of inhibition on tumor growth. Besides, we discussed the possible mechanism of ICA to enhance body immunity by detecting the expression level of MHC-I and related genes in B16-F10 and RMA/S cells. The results suggested that ICA has the potential to up-regulate LMP/TAP related molecules and induce the expression of MHC-I, which increase the immune surveillance and keep cancer in remission. In conclusion, ICA showed an anti-tumor effect both in vitro and in vivo and may be an effective antigen adjuvant for cancer treatment by enhancing tumor-specific immunity.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Flavonoides/farmacología , Inmunoterapia/métodos , Mastocitoma/terapia , Adyuvantes Inmunológicos/administración & dosificación , Animales , Antígenos de Neoplasias/administración & dosificación , Antígenos de Neoplasias/inmunología , Línea Celular Tumoral , Femenino , Flavonoides/administración & dosificación , Antígenos de Histocompatibilidad Clase I/inmunología , Macrófagos Peritoneales/inmunología , Mastocitoma/inmunología , Ratones , Ratones Endogámicos DBA , Bazo/citología , Bazo/inmunología , Linfocitos T Citotóxicos/inmunología , Regulación hacia Arriba/inmunologíaRESUMEN
OBJECTIVE: To evaluate safety and efficacy of LDI-100, a preparation containing human chorionic gonadotropin (hCG) and bacillus Calmette-Guerin (BCG), in the treatment of dogs with mast cell tumors and to compare results with those from a control group receiving single-agent vinblastine. ANIMALS: 95 dogs with measurable grade II or III mast cell tumors. PROCEDURES: Dogs were randomized to receive either LDI-100 (1.35 ng of BCG and 2 units of hCG, SC, q 24 h) or vinblastine (2 mg/m(2), IV, q 1 wk) for 6 weeks. Tumors were measured at baseline and day 42, and dogs were monitored for signs of toxicosis. Clinical performance scores were recorded at each visit. Differences in host factors (sex, weight, and age), clinical performance score, tumor response, and adverse events were analyzed. RESULTS: 46 dogs received LDI-100, and 49 dogs received vinblastine. No significant differences were found between the 2 treatment groups with regard to host factors or clinical performance score. Tumor response (>or=50% reduction) rates were similar between the LDI-100 and vinblastine group (28.6% and 11.7%, respectively). Dogs in the LDI-100 group had significantly less neutropenia than the vinblastine group. CONCLUSIONS AND CLINICAL RELEVANCE: hCG and BCG have immunomodulatory and antitumor effects against a variety of malignancies in humans and dogs. In this study, LDI-100 provided clinical responses comparable to single-agent vinblastine chemotherapy but without myelosuppression. LDI-100 is a promising new agent that should be further investigated for multimodality therapy of mast cell tumors in dogs.
Asunto(s)
Adyuvantes Inmunológicos/uso terapéutico , Vacuna BCG/uso terapéutico , Gonadotropina Coriónica/uso terapéutico , Enfermedades de los Perros/terapia , Mastocitoma/veterinaria , Animales , Antineoplásicos/uso terapéutico , Gonadotropina Coriónica/inmunología , Enfermedades de los Perros/inmunología , Perros , Femenino , Humanos , Masculino , Mastocitoma/inmunología , Mastocitoma/terapia , Estudios Prospectivos , Vinblastina/uso terapéuticoRESUMEN
BACKGROUND: Electrochemotherapy (ECT) is a novel anticancer therapy that combines the delivery of trains of appropriate waveforms with the local administration of chemotherapy agents. The purpose of this investigation was to assess the adjuvant potentials of ECT for the treatment of incompletely excised mast cell tumors (MCT). MATERIALS AND METHODS: Twenty-eight privately-owned dogs with incompletely removed MCT were treated with intralesional bleomycin (1.5 IU/cm2) followed by the application of trains of biphasic pulses (8 pulses, 1300 V/cm, 50 + 50 micros duration, 1 Hz frequency). RESULTS: The overall response rate was 85% with a mean estimated time to recurrence of 52.76 +/- 6.5 months (range: 39.99 to 65.54 months, 95% CI). At the time of writing this report, the median survival time was not reached. Three dogs died of metastatic disease that they developed at the same time of local recurrence, one developed multiple cutaneous nodules at different locations and one with recurrence was re-treated and is currently disease-free after 22 months. No major local or systemic toxicities were noted for the duration of the study. CONCLUSION: ECT is a safe and effective therapy for incompletely excised MCTs in companion animals. Its ease of administration, lack of toxicities and low cost make it an attractive alternative to standard treatments and warrants further investigation.
Asunto(s)
Antineoplásicos/uso terapéutico , Enfermedades de los Perros/terapia , Terapia por Estimulación Eléctrica , Mastocitoma/terapia , Animales , Antineoplásicos/efectos adversos , Terapia Combinada , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/cirugía , Perros , Terapia por Estimulación Eléctrica/efectos adversos , Femenino , Masculino , Mastocitoma/tratamiento farmacológico , Mastocitoma/cirugía , Mastocitoma/veterinaria , RecurrenciaRESUMEN
KIT receptor, the c-kit gene product, is thought to play a major role in canine mastocytoma, one of the most common neoplastic diseases in dogs. In the present study, the expression of c-kit proto-oncogene in blood and in tumor biopsies from 41 dogs with histologically confirmed mastocytoma at different grades of cellular differentiation and 5 negative control dogs was investigated using real-time (quantitative) reverse transcription-polymerase chain reaction (RRT-PCR). The animals were followed up for over 1 year after surgery in order to characterize the kinetics of c-kit expression in blood. Transcript mRNAs extracted from blood at different time points after surgery and from tumor tissue surgically removed from each dog were used in a quantitative RRT-PCR assay targeting the extracellular coding region of the c-kit gene. Tissues constitutively expressing c-kit (brain and spleen) were used as positive controls. Levels of expression of c-kit were higher in tumor biopsies than in blood; the blood level decreased in the patients between 1 and 3 months after surgery. No KIT expression was detected in blood from the 5 dogs not affected by mastocytoma (negative controls). The RRT-PCR appears to be a suitable method for sensitive and quantitative detection of c-kit gene expression in canine blood and neoplastic tissues. Although c-kit expression levels measured by RRT-PCR do not correlate with prognosis, they confirm that surgery remains the main treatment to reduce circulating mastocytes and that circulating mast cells can be detected even in benign highly differentiated forms of mastocytoma such as grade I.
Asunto(s)
Enfermedades de los Perros/genética , Regulación Neoplásica de la Expresión Génica , Mastocitoma/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Proteínas Proto-Oncogénicas c-kit/genética , Animales , Antineoplásicos/uso terapéutico , Perros , Femenino , Masculino , Mastocitoma/genética , Mastocitoma/terapia , Reacción en Cadena de la Polimerasa/métodos , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
ENMD-2076 is an aurora kinase inhibitor that also has multi-target tyrosine kinase inhibitor properties. In this study, the mRNA and the protein expression of aurora-A and aurora-B were evaluated in three canine mast cell tumour cell lines. Dose-dependent cytotoxicity was seen in the cells treated, and it affected the cell cycle with cells in the G2/M phase being selectively killed. The cells were also evaluated for radiosensitivity with/without ENMD-2076, and radiosensitization was seen after 3 Gy and 6 Gy exposures with ENMD-2076 for 48 h. Protein expression of caspase-3 was gradually increased, and the expression intensity was highest at 24 h post irradiation in cells without ENMD-2076 treatment, which indicates that radiation exposure with ENMD-2076-induced cell death faster than radiation treatment alone. Our study results suggest the potential usefulness of treating canine mast cell tumours with aurora kinase inhibitors alone or in conjunction with radiation therapy.
Asunto(s)
Aurora Quinasas/antagonistas & inhibidores , Enfermedades de los Perros/tratamiento farmacológico , Mastocitoma/terapia , Pirazoles/farmacología , Pirimidinas/farmacología , Fármacos Sensibilizantes a Radiaciones/farmacología , Terapia por Rayos X , Animales , Apoptosis/efectos de los fármacos , Aurora Quinasa A/genética , Aurora Quinasa A/metabolismo , Aurora Quinasa B/genética , Aurora Quinasa B/metabolismo , Aurora Quinasas/genética , Aurora Quinasas/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Perros , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión GénicaRESUMEN
Immunosuppression in the tumor microenvironment blunts vaccine-induced immune effectors. PD-1/B7-H1 is an important inhibitory axis in the tumor microenvironment. Our goal in this study was to determine the effect of blocking this inhibitory axis during and following vaccination against breast cancer. We observed that using anti-PD-1 antibody and a multipeptide vaccine (consisting of immunogenic peptides derived from breast cancer antigens, neu, legumain, and ß-catenin) as a combination therapy regimen for the treatment of breast cancer-bearing mice prolonged the vaccine-induced progression-free survival period. This prolonged survival was associated with increase in number of Tc1 and Tc2 CD8 T cells with memory precursor phenotype, CD27+IL-7RhiT-betlo, and decrease in number of PD-1+ dendritic cells (DC) in regressing tumors and enhanced antigen reactivity of tumor-infiltrating CD8 T cells. It was also observed that blockade of PD-1 on tumor DCs enhanced IL-7R expression on CD8 T cells. Taken together, our results suggest that PD-1 blockade enhances breast cancer vaccine efficacy by altering both CD8 T cell and DC components of the tumor microenvironment. Given the recent success of anti-PD-1 monotherapy, our results are encouraging for developing combination therapies for the treatment of patients with cancer in which anti-PD-1 monotherapy alone may be ineffective (i.e., PD-L1-negative tumors).
Asunto(s)
Anticuerpos/inmunología , Anticuerpos/farmacología , Linfocitos T CD8-positivos/efectos de los fármacos , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/farmacología , Memoria Inmunológica/inmunología , Receptor de Muerte Celular Programada 1/inmunología , Animales , Antígenos de Neoplasias/inmunología , Antígeno B7-H1/inmunología , Linfocitos T CD8-positivos/inmunología , Línea Celular Tumoral , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Supervivencia sin Enfermedad , Femenino , Memoria Inmunológica/efectos de los fármacos , Mastocitoma/inmunología , Mastocitoma/terapia , Ratones , Ratones Endogámicos BALB C , Receptores de Interleucina-7/inmunología , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/inmunologíaRESUMEN
Indoleamine 2,3-dioxygenase (IDO) is a rate-limiting enzyme in tryptophan catabolism that plays an important role in the induction of immune tolerance. Its role in graft-versus-tumor effect after allogeneic stem cell transplantation (allo-SCT) remains unclear. Using a murine graft-versus-tumor model of reduced-intensity allo-HSCT followed by donor leukocyte infusion (DLI), we examined the role of IDO inhibition. Two stereoisomers of 1-methyl tryptophan (1-MT), a small-molecule inhibitor of IDO, reduced the growth of inoculated tumor in the mice that received DLI and had higher expression of IDO1 and IFNγ. However, L-1MT, but not D-1MT, mitigated tumor growth in mice that did not receive DLI and did not express IDO1 and IFNγ. Accordingly, both stereoisomers reduced plasma kynurenine concentrations early after DLI and enhanced in vitro cytotoxic lymphocyte function after allogeneic mixed lymphocyte reaction. Furthermore, L-1MT was more efficient in causing direct cytotoxic effects than D-1MT. Our results suggest that IDO inhibition can benefit anti-tumor therapy in the setting of reduced-intensity allo-SCT using DLI.
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Efecto Injerto vs Tumor/fisiología , Factores Inmunológicos/farmacología , Indolamina-Pirrol 2,3,-Dioxigenasa/antagonistas & inhibidores , Mastocitoma/terapia , Proteínas de Neoplasias/antagonistas & inhibidores , Triptófano/análogos & derivados , Aloinjertos , Animales , Trasplante de Médula Ósea , Línea Celular Tumoral , Citotoxicidad Inmunológica , Evaluación Preclínica de Medicamentos , Inducción Enzimática , Efecto Injerto vs Tumor/efectos de los fármacos , Factores Inmunológicos/uso terapéutico , Indolamina-Pirrol 2,3,-Dioxigenasa/genética , Indolamina-Pirrol 2,3,-Dioxigenasa/fisiología , Interferón gamma/biosíntesis , Quinurenina/biosíntesis , Quinurenina/sangre , Transfusión de Leucocitos , Ganglios Linfáticos/enzimología , Prueba de Cultivo Mixto de Linfocitos , Mastocitoma/tratamiento farmacológico , Mastocitoma/enzimología , Mastocitoma/inmunología , Ratones , Ratones Endogámicos C57BL , Proteínas de Neoplasias/fisiología , Quimera por Radiación , Bazo/enzimología , Estereoisomerismo , Factores de Tiempo , Quimera por Trasplante , Triptófano/química , Triptófano/metabolismo , Triptófano/farmacología , Triptófano/uso terapéuticoAsunto(s)
Bienestar del Animal , Enfermedades de los Perros/terapia , Ética Médica , Calidad de Vida , Animales , Perros , Lipoma/terapia , Lipoma/veterinaria , Mastocitoma/terapia , Mastocitoma/veterinaria , Neoplasias de la Tráquea/terapia , Neoplasias de la Tráquea/veterinaria , Traqueotomía/efectos adversos , Traqueotomía/veterinariaRESUMEN
Mast cell tumours are one of the most common neoplasms in dogs. Mutations in the proto-oncogene c-kit, especially internal tandem duplications of exon 11, are considered to play a crucial role in mast cell tumourigenesis. In this report, two cases that suffered from multiple mast cell tumours containing an internal tandem duplication in the primary lesion but not in the secondary lesions are described. This finding indicates the existence of heterogenous c-kit gene mutations in each site of multiple mast cell tumours. Additionally, these results raise the possibility that the contribution of internal tandem duplications in the malignant transformation of mast cells is quite limited. It is proposed that, for clinicians, genetic analysis of several regions of multiple mast cell tumours is necessary for predicting prognosis and tumour response to KIT inhibitors.
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Enfermedades de los Perros/genética , Mastocitoma/veterinaria , Terapia Molecular Dirigida/veterinaria , Proteínas Proto-Oncogénicas c-kit/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-kit/metabolismo , Animales , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/terapia , Perros , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Predisposición Genética a la Enfermedad , Masculino , Mastocitos , Mastocitoma/diagnóstico , Mastocitoma/genética , Mastocitoma/terapia , Mutación/genética , Pronóstico , Proteínas Proto-Oncogénicas c-kit/genéticaRESUMEN
Photodynamic therapy (PDT) involves the intravenous administration of photosensitizers followed by illumination of the tumor with visible light, leading to local production of reactive oxygen species that cause vascular shutdown and tumor cell death. Antitumor immunity is stimulated after PDT because of the acute inflammatory response that involves activation of the innate immune system, leading to stimulation of adaptive immunity. We carried out PDT using benzoporphyrin derivative and 690-nm light after 15 minutes, in DBA/2 mice bearing either the mastocytoma, P815, which expresses the naturally occurring cancer/testis antigen P1A, or the corresponding tumor P1.204 that lacks P1A expression. Tumor cures, significantly higher survival, and rejection of tumor rechallenge were obtained with P815, which were not seen with P1.204 or seen with P815 growing in nude mice. Both CD4 and CD8 T cells had higher levels of intracellular cytokines when isolated from mice receiving PDT of P815 tumors than P1.204 tumors and CD8 T cells from P815-cured mice recognized the peptide epitope of the P1A antigen (LPYLGWLVF) using pentamer staining. Taken together, these findings show that PDT can induce a potent antigen- and epitope-specific immune response against a naturally occurring mouse tumor antigen.
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Inmunidad Adaptativa/inmunología , Antígenos de Neoplasias/inmunología , Luz , Mastocitoma/inmunología , Fotoquimioterapia , Linfocitos T/inmunología , Animales , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Apoptosis , Western Blotting , Proliferación Celular , Citometría de Flujo , Técnicas para Inmunoenzimas , Mastocitoma/metabolismo , Mastocitoma/terapia , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , Ratones Desnudos , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/metabolismo , Linfocitos T/patologíaRESUMEN
Utilizing a clinically relevant haploidentical (HI) murine transplant model, lethally irradiated B6D2F1 (H2K(b/d)) mice were transplanted with T cell-depleted (TCD) BM from B6CBAF1 (H2K(b/k)) mice. We found that administration of IL-15 significantly increases the numbers of CD8+ T and natural killer (NK) cells in spleen and BM after transplantion without GVHD. Graft-versus-tumor (GVT) potency of the graft was evaluated upon tumor challenge using P815 tumor cells (H2(d)). IL-15 administration without T-cell infusion did not result in any survival improvement. However, IL-15 in combination with very low-dose T-cell infusion (1 × 10(4)) significantly increased GVT activity and improved survival in recipients of HI hematopoietic SCT (HSCT). This effect was observed when IL-15 was given at a later time point, rather than immediately following transplantation. IL-15 administration also specifically increased slow-proliferative CD8+ T-cell proliferation and IFN-γ secretion in CD8+ T cells in recipients of CFSE (carboxyfluorescein succinimidyl ester)-labeled HI T-cell infusion, whereas there was no effect on CD4+ T-cell proliferation, suggesting the critical effect of IL-15 on CD8+ T-cell homeostasis in HI host. We conclude that IL-15 can be used for enhancing antileukemia effect of HI-HSCT, which requires presence of donor-derived T cells.