RESUMEN
BACKGROUND: Thiopurines continue to play an important role in the treatment of inflammatory bowel disease (IBD). It is well known that thiopurines can cause several adverse reactions. Especially, hematopoietic toxicity may lead to severe agranulocytosis. In a previous prospective study, we investigated the relationship between inosine triphosphate pyrophosphatase (ITPA) c.94c > a polymorphism, 6-thioguanine nucleotide (6-TGN) concentration and toxicity. METHODS: To clarify the cause of thiopurine toxicity, we analysed nucleoside disphosphate-linked moiety X-type motif 15 (NUDT15) gene polymorphisms, i.e., R139C, V18I, and V19_V19insGV, and measured 6-mercaptopurines and 6-methylmercaptopurines (6-MMP) using the archived blood samples collected from 49 IBD patients for our previous study. RESULTS: The ITPA c.94c > a polymorphism was detected in 19 patients (38.7%, all heterozygous). The R139C polymorphism was found in 10 patients (20.4%, 1 homozygous, 9 heterozygous), V18_V19insGV in 7 patients (14.3%, all heterozygous), and V18I in 2 patients (4.08%, all heterozygous). Although R139C was more strongly associated with leukopenia than c.94c > a, there were no significant correlations with 6-TGN and 6-MMP levels, as for c.94c > a. The leukopenia incidence rates for each gene polymorphism were 0% in those with all wild-type genes, 21.4% for c.94c > a only, 42.9% for NUDT15 polymorphism (s) only, and 80.0% for both polymorphisms. CONCLUSIONS: All cases of leukopenia were associated with ITPA c.94c > a and/or polymorphism of NUDT15 and the risk of developing leukopenia was synergistically increased by ITPA and NUDT15 gene polymorphism. However, there was no association between the level of azathioprine metabolites and these polymorphisms.
Asunto(s)
Azatioprina , Enfermedades Inflamatorias del Intestino , Leucopenia , Pirofosfatasas , Humanos , Azatioprina/efectos adversos , Azatioprina/uso terapéutico , Pueblos del Este de Asia , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/genética , Leucopenia/inducido químicamente , Leucopenia/genética , Mercaptopurina/efectos adversos , Pirofosfatasas/genéticaRESUMEN
BACKGROUND: We have previously reported that patients with Crohn's disease (CD) have a very specific erythrocyte membrane phospholipid fatty acid profile. The findings of this study suggest that the activities of enzymes involved in the metabolism of linoleic acid (LA), that is, delta-6 desaturase, are higher in CD patients than in healthy individuals. METHODS: We evaluated the utilities of various fatty acid compositions of the plasma (p-) as new serological markers for CD compared to those of erythrocyte membranes (e-). RESULTS: Fifty CD patients and 50 healthy individuals were enrolled. In both plasma and erythrocyte membranes, the weight percentages of palmitic acid (PA) were significantly higher, while those of LA were significantly lower in CD patients than in controls. Fatty acids with high sensitivity and specificity were p-PA (0.86 and 0.74) and e-PA (0.80 and 0.74). With PA and LA as a CD fatty acid index (CDFAi), that is, CDFAi = (PA/LA), the sensitivity and specificity of plasma CDFAi (p-CDFAi) and e-CDFAi were 0.80 and 0.80; and 0.82 and 0.88 respectively. CONCLUSION: In CD patients, various fatty acids were specifically altered in both plasma and erythrocytes, and p-PA and p-CDFAi are potentially useful as new serological markers for CD.
Asunto(s)
Enfermedad de Crohn/sangre , Ácidos Grasos/sangre , Adulto , Biomarcadores/sangre , Estudios de Casos y Controles , Membrana Eritrocítica/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fosfolípidos/sangre , Curva ROC , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: Several studies have suggested that an elevated neutrophil-lymphocyte ratio (NLR) is associated with a poorer prognosis in patients with pancreatic cancer (PC). The correlations between the NLR and immunohistochemical (IHC) analysis with regard to the prognosis of patients with PC remain to be elucidated. By using IHC findings, we determined the value of the NLR as a prognostic factor in patients with PC. MATERIAL AND METHODS: We collected the clinico-pathological data of 28 consecutive patients who underwent surgical resection for PC between January 2008 and December 2012 at The Jikei University Kashiwa Hospital. We investigated whether the NLR and IHC results were related and ensured the consistency of the prognosis of patients with PC. RESULTS: The Kaplan-Meier curves for the disease-free survival (DFS) and the overall survival (OS) revealed that an NLR ≥ 5 is an implicit factor for decreased DFS and OS in patients with PC (p = 0.003, p < 0.001, log-rank test). The density of CD163(+) macrophages and CD66b(+) neutrophils was significantly higher in the high NLR group; on the contrary, the density of CD20(+) lymphocytes was significantly higher in the low NLR group. Moreover, a Mann-Whitney U test showed that the NLR was significantly correlated with a high density of CD20(+) lymphocytes (p = 0.031) and CD163(+) macrophages (p = 0.023), while the NLR was not significantly correlated with CD66b(+) neutrophils (p = 0.397). CONCLUSIONS: Our results demonstrated the validity of the NLR by IHC analyses and we determined that a higher value of NLR is a trustworthy prognostic factor for patients with PC.
Asunto(s)
Linfocitos/patología , Neutrófilos/patología , Neoplasias Pancreáticas/sangre , Medición de Riesgo/métodos , Carga Tumoral , Anciano , Supervivencia sin Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Inmunohistoquímica , Japón/epidemiología , Recuento de Leucocitos , Masculino , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/mortalidad , Pronóstico , Estudios Retrospectivos , Tasa de Supervivencia/tendenciasRESUMEN
AIM: To investigate the association of plasma levels of interleukin (IL)-6 and -8 with Wilms' tumor 1 (WT1)-specific immune responses and clinical outcomes in patients with pancreatic ductal adenocarcinoma (PDA) treated with dendritic cells (DCs) pulsed with three types of major histocompatibility complex class I and II-restricted WT1 peptides combined with chemotherapy. METHODS: During the entire treatment period, plasma levels of IL-6 and -8 were analyzed by ELISA. The induction of WT1-specific immune responses was assessed using the WT1 peptide-specific delayed-type hypersensitivity (DTH) test. RESULTS: Three of 7 patients displayed strong WT1-DTH reactions throughout long-term vaccination with significantly decreased levels of IL-6/-8 after vaccinations compared with the levels prior to treatment. Moreover, overall survival (OS) was significantly longer in PDA patients with low plasma IL-6 levels (< 2 pg/mL) after 5 vaccinations than in patients with high plasma IL-6 levels (≥ 2 pg/mL) (P = 0.025). After disease progression, WT1-DTH reactions decreased severely and were ultimately negative at the terminal stage of cancer. The decreased levels of IL-6/-8 observed throughout long-term vaccination were associated with WT1-specific DTH reactions and long-term OS. CONCLUSION: Prolonged low levels of plasma IL-6/-8 in PDA patients may be a prognostic marker for the clinical outcomes of chemoimmunotherapy.
Asunto(s)
Antimetabolitos Antineoplásicos/administración & dosificación , Biomarcadores de Tumor/sangre , Carcinoma Ductal Pancreático/tratamiento farmacológico , Células Dendríticas/trasplante , Desoxicitidina/análogos & derivados , Inmunoterapia/métodos , Interleucina-6/sangre , Interleucina-8/sangre , Neoplasias Pancreáticas/tratamiento farmacológico , Adulto , Anciano , Antimetabolitos Antineoplásicos/efectos adversos , Carcinoma Ductal Pancreático/sangre , Carcinoma Ductal Pancreático/inmunología , Carcinoma Ductal Pancreático/mortalidad , Carcinoma Ductal Pancreático/patología , Células Cultivadas , Quimioterapia Adyuvante , Células Dendríticas/inmunología , Desoxicitidina/administración & dosificación , Desoxicitidina/efectos adversos , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Pruebas Inmunológicas , Inmunoterapia/efectos adversos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/mortalidad , Neoplasias Pancreáticas/patología , Fragmentos de Péptidos/inmunología , Factores de Tiempo , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Proteínas WT1/inmunología , GemcitabinaRESUMEN
Azathioprine (AZA) is frequently used in patients with inflammatory bowel disease (IBD). However, toxic adverse reactions frequently develop and limit the clinical benefits. Currently, the precise mechanisms underlying thiopurine-related toxicity are not well understood. To investigate the relationship between the extent of thiopurine metabolism and adverse reactions in Japanese IBD patients, we prospectively observed 48 IBD patients who received AZA. We analyzed the thiopurine S-methyltransferase (TPMT) and inosine triphosphate pyrophosphatase (ITPA) gene mutations and measured the concentrations of 6-thioguanine nucleotide (6-TGN) continuously for 52 weeks. All patients possessed wild-type TPMT gene sequences. The ITPA 94C>A mutation was detected in 19 patients (39.6%). Adverse reactions developed in 14 of the 48 patients (29.2%), including leukopenia in 10 patients (20.8%). In the leukopenia group, the percentages of patients with 94C>A were higher than those in the without-leukopenia group (70.0% vs. 31.6%, P < 0.05). The average concentrations of 6-TGN in the patients with 94C>A were generally higher than those in the patients without 94C>A, however, there were no significant differences. Only 3 out of 10 patients with leukopenia exhibited high 6-TGN levels (30.0%). No negative correlations between white blood cell (WBC) counts and 6-TGN concentrations were observed. The cumulative incidence of leukopenia were higher for patients with 94C>A. Seven out of 19 patients (36.8%) with the ITPA 94C>A mutation developed leukopenia; however, this mutation may not unequivocally increase the risk of developing leukopenia. In addition, there are factors other than increased 6-TGN levels that are involved in the onset of leukopenia.
Asunto(s)
Azatioprina/efectos adversos , Azatioprina/metabolismo , Inmunosupresores/efectos adversos , Inmunosupresores/metabolismo , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/metabolismo , Adulto , Alelos , Eritrocitos/metabolismo , Femenino , Genotipo , Nucleótidos de Guanina/metabolismo , Humanos , Incidencia , Enfermedades Inflamatorias del Intestino/genética , Japón , Leucopenia/epidemiología , Leucopenia/etiología , Masculino , Metiltransferasas/genética , Metiltransferasas/metabolismo , Persona de Mediana Edad , Mutación , Farmacogenética , Estudios Prospectivos , Pirofosfatasas/genética , Pirofosfatasas/metabolismo , Tionucleótidos/metabolismo , Adulto JovenRESUMEN
BACKGROUND/AIM: Chemoimmunotherapy has been used to treat intrahepatic cholangiocarcinoma (ICC). However, little is known about the phenomena underlying the immunomodulation of ICC cells elicited by chemoimmunotherapy. MATERIALS AND METHODS: Primary ICC cells from a patient with ICC who received gemcitabine followed by 5-fluorouracil (5-FU), both combined with dendritic cells pulsed with Wilms' tumor 1 (WT1) peptides were cultured. ICC cells were treated with gemcitabine, 5-FU or interferon (IFN)-γ in vitro. The phenotype of the ICC cells was examined by flow cytometry and quantitative reverse transcription polymerase chain reaction. RESULTS: Stimulation of the ICC cells with gemcitabine resulted in up-regulation of WT1 mRNA, programmed death receptor ligand-1 (PDL1) and calreticulin. Gemcitabine, 5-FU and IFN-γ induced up-regulation of mucin-1. Moreover, human leukocyte antigen (HLA)-ABC, HLA-DR and PDL1 were extremely up-regulated by IFN-γ. CONCLUSION: Chemoimmunomodulating agents alter the immunogenicity of ICC cells, resulting in complex clinical efficacy results.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias de los Conductos Biliares/terapia , Conductos Biliares Intrahepáticos/inmunología , Colangiocarcinoma/terapia , Células Dendríticas/inmunología , Inmunoterapia , Antígenos de Neoplasias/inmunología , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Neoplasias de los Conductos Biliares/inmunología , Neoplasias de los Conductos Biliares/patología , Conductos Biliares Intrahepáticos/efectos de los fármacos , Conductos Biliares Intrahepáticos/patología , Colangiocarcinoma/inmunología , Colangiocarcinoma/patología , Terapia Combinada , Desoxicitidina/administración & dosificación , Desoxicitidina/análogos & derivados , Femenino , Fluorouracilo/administración & dosificación , Antígenos HLA-DR/genética , Antígenos HLA-DR/metabolismo , Humanos , Interferón gamma/administración & dosificación , Metástasis Linfática , Persona de Mediana Edad , Mucina-1/genética , Mucina-1/metabolismo , Neoplasias Peritoneales/inmunología , Neoplasias Peritoneales/secundario , Neoplasias Peritoneales/terapia , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Proteínas WT1/genética , Proteínas WT1/metabolismo , GemcitabinaRESUMEN
PURPOSE: We performed a phase I trial to investigate the safety, clinical responses, and Wilms' tumor 1 (WT1)-specific immune responses following treatment with dendritic cells (DC) pulsed with a mixture of three types of WT1 peptides, including both MHC class I and II-restricted epitopes, in combination with chemotherapy. EXPERIMENTAL DESIGN: Ten stage IV patients with pancreatic ductal adenocarcinoma (PDA) and 1 patient with intrahepatic cholangiocarcinoma (ICC) who were HLA-positive for A*02:01, A*02:06, A*24:02, DRB1*04:05, DRB1*08:03, DRB1*15:01, DRB1*15:02, DPB1*05:01, or DPB1*09:01 were enrolled. The patients received one course of gemcitabine followed by biweekly intradermal vaccinations with mature DCs pulsed with MHC class I (DC/WT1-I; 2 PDA and 1 ICC), II (DC/WT1-II; 1 PDA), or I/II-restricted WT1 peptides (DC/WT1-I/II; 7 PDA), and gemcitabine. RESULTS: The combination therapy was well tolerated. WT1-specific IFNγ-producing CD4(+) T cells were significantly increased following treatment with DC/WT1-I/II. WT1 peptide-specific delayed-type hypersensitivity (DTH) was detected in 4 of the 7 patients with PDA vaccinated with DC/WT1-I/II and in 0 of the 3 patients with PDA vaccinated with DC/WT1-I or DC/WT1-II. The WT1-specific DTH-positive patients showed significantly improved overall survival (OS) and progression-free survival (PFS) compared with the negative control patients. In particular, all 3 patients with PDA with strong DTH reactions had a median OS of 717 days. CONCLUSIONS: The activation of WT1-specific immune responses by DC/WT1-I/II combined with chemotherapy may be associated with disease stability in advanced pancreatic cancer.
Asunto(s)
Células Dendríticas/inmunología , Desoxicitidina/análogos & derivados , Epítopos/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Neoplasias Pancreáticas/terapia , Proteínas WT1/inmunología , Adenocarcinoma/inmunología , Adenocarcinoma/mortalidad , Adenocarcinoma/secundario , Adenocarcinoma/terapia , Adulto , Anciano , Antimetabolitos Antineoplásicos/uso terapéutico , Neoplasias de los Conductos Biliares/inmunología , Neoplasias de los Conductos Biliares/mortalidad , Neoplasias de los Conductos Biliares/secundario , Neoplasias de los Conductos Biliares/terapia , Conductos Biliares Intrahepáticos/inmunología , Biomarcadores de Tumor/análisis , Linfocitos T CD8-positivos/inmunología , Carcinoma Ductal Pancreático/inmunología , Carcinoma Ductal Pancreático/mortalidad , Carcinoma Ductal Pancreático/secundario , Carcinoma Ductal Pancreático/terapia , Colangiocarcinoma/inmunología , Colangiocarcinoma/mortalidad , Colangiocarcinoma/secundario , Colangiocarcinoma/terapia , Terapia Combinada , Desoxicitidina/uso terapéutico , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/mortalidad , Neoplasias Pancreáticas/patología , Fragmentos de Péptidos/inmunología , Pronóstico , Tasa de Supervivencia , Linfocitos T Citotóxicos/inmunología , Vacunación , GemcitabinaRESUMEN
Gemcitabine application for patients with impaired renal function or undergoing haemodialysis will increase if the efficacy and safety are proved as the treatment for pancreatic cancer of these patients. However, there is no guideline about the usage of gemcitabine in patients with impaired renal function or haemodialysis. We report the case of a 70-year-old man with advanced pancreatic cancer undergoing haemodialysis. After discontinuation of 100% or 80% dosage, 60% dose of gemcitabine was administered biweekly. Serum carbohydrate antigen 19-9 and carcinoembryonic antigen levels were marked by slight variations and abdominal computed tomography (CT) showed the tumour size hardly changed. We administered gemcitabine for the patient 14 times in total, and he survived over 8 months from the definitive diagnosis. These findings confirm the efficacy and safety of treatment with a biweekly 60% dose of gemcitabine for patients with advanced pancreatic cancer undergoing haemodialysis in the face of dose modification.
Asunto(s)
Antimetabolitos Antineoplásicos/administración & dosificación , Desoxicitidina/análogos & derivados , Neoplasias Pancreáticas/tratamiento farmacológico , Diálisis Renal , Anciano , Desoxicitidina/administración & dosificación , Humanos , Masculino , Tomografía Computarizada por Rayos X , GemcitabinaRESUMEN
Previous work has demonstrated that intestinal bacteria, such as Fusobacterium varium (F. varium), contribute to the clinical activity in ulcerative colitis (UC); thus, an antibiotic combination therapy (amoxicillin, tetracycline, and metronidazole (ATM)) against F. varium can induce and maintain UC remission. Therefore, we investigated whether ATM therapy induces a long-term alteration of intestinal microbiota in patients with UC. Patients with UC were enrolled in a multicenter, randomized, double-blind, placebo-controlled study. Biopsy samples at the beginning of the trial and again at 3 months after treatment completion were randomly obtained from 20 patients. The terminal restriction fragment length polymorphism (T-RFLP) in mucosa-associated bacterial components was examined to assess the alteration of the intestinal microbiota. Profile changes of T-RFLP in mucosa-associated bacterial components were found in 10 of 12 patients in the treatment group and in none of 8 in the placebo group. Dice similarity coefficients using the unweighted pair group method with arithmetic averages (Dice-UPGMA) confirmed that the similarity of mucosal microbiota from the descending colon was significantly decreased after the ATM therapy, and this change was maintained for at least 3 months. Moreover, at 3 months after treatment completion, the F. varium/ß-actin ratio, examined by real-time PCR using nested PCR products from biopsy samples, was reduced less than 40% in 8 of 12 treated patients, which was higher, but not significantly, than in 4 of 8 patients in the placebo group. Together, these results suggest that ATM therapy induces long-term alterations in the intestinal microbiota of patients with UC, which may be associated, at least in part, with clinical effects of the therapy.
Asunto(s)
Colitis Ulcerosa/microbiología , Infecciones por Fusobacterium/microbiología , Fusobacterium/genética , Mucosa Intestinal/microbiología , Microbiota/genética , Actinas/metabolismo , Adolescente , Adulto , Anciano , Amoxicilina/uso terapéutico , Antibacterianos/uso terapéutico , Colitis Ulcerosa/tratamiento farmacológico , Método Doble Ciego , Quimioterapia Combinada , Femenino , Fusobacterium/aislamiento & purificación , Infecciones por Fusobacterium/tratamiento farmacológico , Humanos , Mucosa Intestinal/efectos de los fármacos , Masculino , Metronidazol/uso terapéutico , Persona de Mediana Edad , Tetraciclina/uso terapéuticoAsunto(s)
Colitis Ulcerosa/microbiología , Edwardsiella tarda/aislamiento & purificación , Infecciones por Enterobacteriaceae/microbiología , Sobreinfección/microbiología , Adulto , Antibacterianos/uso terapéutico , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/patología , Infecciones por Enterobacteriaceae/tratamiento farmacológico , Heces/microbiología , Femenino , Humanos , Levofloxacino/uso terapéutico , Masculino , Persona de Mediana Edad , Recurrencia , Índice de Severidad de la Enfermedad , Sobreinfección/tratamiento farmacológico , Adulto JovenRESUMEN
AIM: To investigate the association of procalcitonin (PCT) with ulcerative colitis (UC) activity. METHODS: Serum PCT levels, C-reactive protein (CRP) levels, the erythrocyte sedimentation rate, and the white blood cell count were analyzed in 18 patients with UC and 11 healthy volunteers. Serum PCT levels were analyzed by an electrochemiluminescence immunoassay. Severity assessments were based on Truelove and Witts' severity index. Correlation of serum PCT and CRP levels with UC activity was examined. Moreover, we assessed serum PCT and CRP levels in patients with a Mayo endoscopic subscore. RESULTS: Serum PCT levels in severe UC patients (n = 7) (0.096 ± 0.034 ng/mL) were significantly higher than in mild-to-moderate UC patients (n = 11) (0.033 ± 0.012 ng/mL) and healthy volunteers (n = 11) (0.035 ± 0.005 ng/mL) (P = 0.0005 and P < 0.0001, respectively). In addition, there was no difference in serum PCT levels between mild-to-moderate UC patients and healthy volunteers. Interestingly, patients with a Mayo endoscopic subscore of 3 points displayed significantly increased levels of serum PCT (0.075 ± 0.043 ng/mL) compared with patients with a subscore of 2 points (0.03 ± 0.011 ng/mL) (P = 0.0302). Moreover, CRP levels in patients with severe UC or a Mayo endoscopic subscore of 3 points were not significantly higher than in patients with mild-to-moderate UC or a Mayo endoscopic subscore of 3 points. CONCLUSION: Serum PCT levels were significantly correlated with UC activity.
Asunto(s)
Calcitonina/sangre , Colitis Ulcerosa/sangre , Precursores de Proteínas/sangre , Adulto , Biomarcadores/sangre , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Péptido Relacionado con Gen de Calcitonina , Estudios de Casos y Controles , Colitis Ulcerosa/patología , Colonoscopía , Femenino , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Estudios Prospectivos , Índice de Severidad de la Enfermedad , Regulación hacia Arriba , Adulto JovenRESUMEN
The therapeutic efficacy of fusion cell (FC)-based cancer vaccine generated with whole tumor cells and dendritic cells (DCs) requires the improved immunogenicity of both cells. Treatment of whole tumor cells with ethanol resulted in blockade of immune-suppressive soluble factors such as transforming growth factor (TGF)-ß1, vascular endothelial growth factor, and IL-10 without decreased expression of major histocompatibility complex (MHC) class I and the MUC1 tumor-associated antigen. Moreover, the ethanol-treated tumor cells expressed "eat-me" signals such as calreticulin (CRT) on the cell surface and released immunostimulatory factors such as heat shock protein (HSP)90α and high-mobility group box 1 (HMGB1). A dual stimulation of protein-bound polysaccharides isolated from Coriolus versicolor (TLR2 agonist) and penicillin-inactivated Streptococcus pyogenes (TLR4 agonist) led human monocyte-derived DCs to produce HSP90α and multiple cytokines such as IL-12p70 and IL-10. Interestingly, incorporating ethanol-treated tumor cells and TLRs-stimulated DCs during the fusion process promoted fusion efficiency and up-regulated MHC class II molecules on a per fusion basis. Moreover, fusions of ethanol-treated tumor cells and dual TLRs-stimulated DCs (E-tumor/FCs) inhibited the production of multiple immune-suppressive soluble factors including TGF-ß1 and up-regulated the production of IL-12p70 and HSP90α. Most importantly, E-tumor/FCs activated T cells capable of producing high levels of IFN-γ, resulting in augmented MUC1-specific CTL induction. Collectively, our results illustrate the synergy between ethanol-treated whole tumor cells and dual TLRs-stimulated DCs in inducing augmented CTL responses in vitro by FC preparations. The alternative system is simple and may provide a platform for adoptive immunotherapy.
Asunto(s)
Células Dendríticas/inmunología , Inmunoterapia Adoptiva , Interleucina-12/biosíntesis , Neoplasias/terapia , Receptores Toll-Like/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Fusión Celular , Línea Celular Tumoral , Citocinas/metabolismo , Etanol/farmacología , Humanos , Factores Inmunológicos/metabolismo , Activación de Linfocitos , Mucina-1/inmunología , Neoplasias/inmunología , Fragmentos de Péptidos/inmunología , Fenotipo , Linfocitos T Citotóxicos/inmunología , Receptores Toll-Like/agonistasRESUMEN
Induction of antitumor immunity by dendritic cell (DC)-tumor fusion cells (DC/tumor) can be modulated by their activation status. In this study, to address optimal status of DC/tumor to induce efficient antigen-specific cytotoxic T lymphocytes (CTLs), we have created various types of DC/tumor: 1) un-activated DC/tumor; 2) penicillin-killed Streptococcus pyogenes (OK-432; TLR4 agonist)-activated DC/tumor; 3) protein-bound polysaccharides isolated from Coriolus versicolor (PSK; TLR2 agonist)-activated DC/tumor; and 4) Combined OK-432- and PSK-activated DC/tumor. Moreover, we assessed the effects of TGF-ß1 derived from DC/tumor on the induction of MUC1-specific CTLs. Combined TLR2- and TLR4-activated DC/tumor overcame immune-suppressive effect of TGF-ß1 in comparison to those single activated or un-activated DC/tumor as demonstrated by: 1) up-regulation of MHC class II and CD86 expression on DC/tumor; 2) increased fusion efficiency; 3) increased production of fusions derived IL-12p70; 4) activation of CD4(+) and CD8(+) T cells that produce high levels of IFN-γ; 5) augmented induction of CTL activity specific for MUC1; and 6) superior efficacy in inhibiting CD4(+)CD25(+)Foxp3(+) T cell generation. However, DC/tumor-derived TGF-ß1 reduced the efficacy of DC/tumor vaccine in vitro. Incorporating combined TLRs-activation and TGF-ß1-blockade of DC/tumor may enhance the effectiveness of DC/tumor-based cancer vaccines and have the potential applicability to the field of adoptive immunotherapy.
Asunto(s)
Células Dendríticas/inmunología , Linfocitos T Citotóxicos/inmunología , Receptor Toll-Like 2/agonistas , Receptor Toll-Like 4/agonistas , Factor de Crecimiento Transformador beta1/farmacología , Antígeno B7-2/genética , Antígeno B7-2/inmunología , Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/inmunología , Fusión Celular , Línea Celular Tumoral , Células Dendríticas/citología , Células Dendríticas/efectos de los fármacos , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interferón gamma/biosíntesis , Interferón gamma/inmunología , Interleucina-12/biosíntesis , Interleucina-12/inmunología , Activación de Linfocitos/efectos de los fármacos , Mucina-1/genética , Mucina-1/inmunología , Picibanil/farmacología , Proteoglicanos/farmacología , Transducción de Señal , Linfocitos T Citotóxicos/citología , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunología , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Fusobacterium varium is an elusive pathogenic factor in ulcerative colitis (UC); conventional methods of fecal culture rarely recover F. varium. We have developed a nested culture method to recover Fusobacterium and we used it to investigate whether F. varium could be isolated from UC patients. We enrolled 50 consecutive patients in this study; 26 received combination antibiotic therapy that included amoxicillin, tetracycline, and metronidazole (ATM) for 2 weeks and were thus assigned to the ATM group, and the remaining 24 were assigned to the non-ATM group and did not receive any antibiotics. Stool samples were added to 10 ml of GAM broth that contained neomycin and crystal violet. The samples were vortexed and incubated under anaerobic conditions. The preincubated broth was streaked onto a Fusobacterium-selective agar plate and then incubated under anaerobic conditions. The species of the colonies isolated were identified using the Vitek Automated system and PCR analysis. We recoverd F. varium from 7 of the 24 non-ATM patients (29.2%) and none from the ATM patients (0%) (P = 0.0035). All of the F. varium isolates were susceptible to ATM. This study suggests that the recovery of F. varium is related to UC, which aligns with results from previous studies that used mucosal culture, immunostaining, real-time PCR, and serological studies.
Asunto(s)
Técnicas Bacteriológicas/métodos , Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/microbiología , Heces/microbiología , Fusobacterium/aislamiento & purificación , Adulto , Anciano , Técnicas de Tipificación Bacteriana , Medios de Cultivo/química , Femenino , Fusobacterium/crecimiento & desarrollo , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , Adulto JovenRESUMEN
BACKGROUND AND OBJECTIVES: The sudden change in the dietary habits of the Japanese population towards a European/American-style diet since the 1960s is thought to be responsible for the recent increase in the incidence of inflammatory bowel disease (IBD) in Japan. Dietary fatty acid intake influences the fatty acid profiles of vital cell membranes, which might be a source of inflammatory mediators. METHODS: We investigated the fatty acid composition of the erythrocyte membrane in 90 healthy Japanese and 43 initial-onset IBD patients (ulcerative colitis, UC: 25; Crohn's disease, CD: 18) who had not undergone any dietary intervention to examine the role fatty acids play in the onset of IBD. RESULTS: The erythrocyte membrane n-3/n-6 ratio of the initial-onset IBD patients was 0.42 ± 0.13, which was not significantly different from that of the healthy Japanese subjects (0.41 ± 0.13). However, the CD patients displayed a significantly lower mean percentage weight (MPW) of linoleic acid (LA) than the healthy subjects (8.25 ± 1.75 vs. 9.90 ± 1.29; p < 0.001), while their MPW of arachidonic acid (AA) was significantly higher than those of the healthy subjects and UC patients (11.22 ± 2.18 vs. 9.76 ± 1.64, p < 0.01; vs. 9.58 ± 1.97, p < 0.01, respectively). The mean delta 6-desaturation index of the CD patients was significantly higher than that of the healthy subjects (1.61 ± 0.65 vs. 1.11 ± 0.26; p < 0.001). CONCLUSIONS: The CD patients displayed significantly higher and lower MPW of AA and LA, respectively, than the healthy subjects, suggesting that delta 6-desaturase is hyperactivated in CD. The cell membrane fatty acid profile might be a therapeutic target in CD.
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Membrana Eritrocítica/metabolismo , Ácidos Grasos/metabolismo , Enfermedades Inflamatorias del Intestino/metabolismo , Fosfolípidos/metabolismo , Adulto , Pueblo Asiatico , Estudios de Casos y Controles , Femenino , Humanos , Japón , Linoleoil-CoA Desaturasa/metabolismo , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Pancreatic cancer is a highly aggressive and notoriously difficult to treat. As the vast majority of patients are diagnosed at advanced stage of the disease, only a small population is curative by surgical resection. Although gemcitabine-based chemotherapy is typically offered as standard of care, most patients do not survive longer than 6 months. Thus, new therapeutic approaches are needed. Pancreatic cancer cells that develop gemcitabine resistance would still be suitable targets for immunotherapy. Therefore, one promising treatment approach may be immunotherapy that is designed to target pancreatic-cancer-associated antigens. In this paper, we detail recent work in immunotherapy and the advances in concept of combination therapy of immunotherapy and chemotherapy. We offer our perspective on how to increase the clinical efficacy of immunotherapies for pancreatic cancer.
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Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer , Quimioterapia , Inmunoterapia , Neoplasias Pancreáticas/tratamiento farmacológico , Animales , Linfocitos T CD8-positivos/inmunología , Terapia Combinada , Citotoxicidad Inmunológica , Células Dendríticas/inmunología , Humanos , Inmunoterapia/tendencias , Activación de Linfocitos , Neoplasias Pancreáticas/inmunología , Resultado del Tratamiento , Escape del TumorRESUMEN
BACKGROUND: N-3 polyunsaturated fatty acids (PUFA) are considered important pharmaconutrients for modulating mucosal immunity and therapeutic responses in patients with inflammatory bowel disease (IBD). We investigated the influence of diet therapy involving the use of an "n-3 PUFA food exchange table" (n-3DP) on the fatty acid composition of the erythrocyte membranes of IBD patients and its remission-maintaining effects. METHODS: We analyzed the fatty acid composition of the erythrocyte membrane before and after n-3DP intervention in 20 initial-onset IBD patients who had not undergone any dietary intervention. We then analyzed it again and evaluated disease activity after 12-18 months intervention in 230 IBD patients (168 ulcerative colitis, 62 Crohn's disease; follow-up group) in whom n-3DP was introduced after remission had been achieved. The follow-up group was divided into remission and relapse groups. RESULTS: In the 20 initial-onset patients, the mean n-3/n-6 ratio significantly increased after intervention (0.41 ± 0.16 versus 0.70 ± 0.20; P < 0.001). In the follow-up group the ratio in the remission group (n = 145) was significantly higher than that in the relapse group (n = 85) (0.65 ± 0.28 versus 0.53 ± 0.18; P < 0.001). The ratio significantly decreased in those who suffered a relapse after the beginning of treatment (P < 0.01). CONCLUSIONS: N-3DP significantly increased the erythrocyte membrane n-3/n-6 ratio in IBD patients, and this ratio was significantly higher in the remission group, suggesting that n-3DP alters the fatty acid composition of the cell membrane and influences clinical activity in IBD patients.
Asunto(s)
Colitis Ulcerosa/dietoterapia , Enfermedad de Crohn/dietoterapia , Ácidos Grasos Omega-3/administración & dosificación , Adulto , Colitis Ulcerosa/sangre , Enfermedad de Crohn/sangre , Dieta , Membrana Eritrocítica/metabolismo , Ácidos Grasos Omega-3/metabolismo , Femenino , Estudios de Seguimiento , Humanos , Masculino , PronósticoRESUMEN
Marshall and Warren were the first to succeed in culturing Helicobacter pylori (H. pylori) from the gastric mucosa of patients with gastritis in 1983. H. pylori is a spiral-shaped bacterium that resides in the gastric mucosa and is one of the most common infections worldwide. H. pylori infection causes gastritis and peptic ulcers and is associated with the development of gastric cancer and MALT lymphoma. Now, a variety of accurate diagnostic tests are widely available. Both invasive tests (bacterial culture, histopathology, and RUT) and non-invasive tests (UBT and serological test) are conducted for the diagnosis of H. pylori infection. This review provides a general overview of the diagnostic methods and tests the characteristics (sensitivity and specificity) for H. pylori infection.