RESUMEN
In order to determine the molecular basis of cytoplasmic male sterility (CMS) in alloplasmic lines of eggplant, the genomic structures and transcription patterns of mitochondrial ATP synthase subunit (atp) and cytochrome oxidase subunit (cox) genes were studied for wild and cultivated eggplants. Alloplasmic eggplant lines with cytoplasms of wild Solanum species showing either anther indehiscent type of CMS or non-pollen production type of CMS were studied with the cultivated eggplant Solanum melongena, used as a control. Southern hybridization of the mitochondrial genes indicated the difference between the two types of CMS and showed complete identity within each type. The cytoplasmic patterns of all wild species differed from that of the cultivated eggplant. Thus, the cytoplasm of the six wild eggplants and the one cultivated eggplant was classified into three groups. Male sterile plants of both types of CMS showed novel transcription patterns of atp1, whereas a different transcription pattern of cox2 was observed only in the anther indehiscent type. Based on these differences, we determined the DNA sequences of about a 4 kbp segment in the atp1 region. Although the coding and 3' flanking regions were almost identical among the cytoplasms, the 5' flanking region was completely different and novel open reading frames (orfs) were found for each of the CMS types and the cultivated eggplant. The cytoplasm of Solanum kurzii inducing the anther indehiscent type CMS had orf312, and those of Solanum aethiopicum and Solanum grandifolium of non-pollen production type CMS had orf218. The correspondence between the transcription patterns of these orfs and phenotypic expression of male sterility strongly suggests that these orfs are causal genes for each type of CMS.
Asunto(s)
Genes Mitocondriales , Genes de Plantas , Infertilidad Vegetal/genética , Solanum/genética , Transcripción Genética , Secuencia de Bases , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADNRESUMEN
Little is known about the correlation between genotype and drug susceptibility in Mycobacterium avium (Mav) strains isolated from patients with Mav infections. To examine whether drug susceptibility profile of Mav is associated with genotype, we carried out variable-number tandem-repeat (VNTR) typing and drug susceptibility testing for Mav isolates from Japanese with nodular-bronchiectasis (NB)-type and cavitary disease (CA)-type diseases. We performed M. avium tandem repeat (MATR)-VNTR typing and drug susceptibility testing by the broth dilution method, using macrolides, rifamycins, ethambutol, isoniazid, aminoglycosides, and quinolones, for Mav isolates from patients with NB and CA-type diseases (NB-Mav and CA-Mav). Based on the VNTR genotyping, the Mav strains were grouped into three clusters. There was no difference with respect to the distribution of NB-Mav and CA-Mav among the clusters. We observed a strong association between VNTR genotype and susceptibility to quinolones (levofloxacin, moxifloxacin, gatifloxacin, sitafloxacin, and garenoxacin) and ethambutol. There was essentially no significant difference in drug susceptibility between NB- and CA-Mav strains, although NB-Mav was somewhat more resistant to fluoroquinolones, especially gatifloxacin, than CA-Mav. There was a significant association between VNTR genotype and susceptibility to quinolones and ethambutol in Mav isolates from Japanese patients.
Asunto(s)
Antituberculosos/farmacología , Repeticiones de Minisatélite , Tipificación Molecular , Mycobacterium avium/clasificación , Mycobacterium avium/efectos de los fármacos , Tuberculosis/microbiología , Anciano , Anciano de 80 o más Años , Análisis por Conglomerados , Femenino , Genotipo , Humanos , Japón , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Mycobacterium avium/genética , Mycobacterium avium/aislamiento & purificaciónRESUMEN
Mycobacterium avium (Mav) lung infections, called nodular-bronchiectasis (NB)-type M. avium complex (MAC) disease, are globally increasing. To elucidate whether there are unusual populations of Mav, causing NB-type disease rather than cavitary (CA)-type disease, we compared the virulence of Mav isolates from patients with NB-type (NB-Mav) and those from CA-type (CA-Mav) diseases, based on intracellular growth in various types of human cells. Five strains each of NB-Mav and CA-Mav were compared with each other for their invasiveness and ability to intracellularly replicate in various types of cultured cells of human origin. The two types of Mav isolates showed a similar ability, on average, to replicate in macrophages and lung epithelial cells. Moreover, they showed a similar ability to induce the production of reactive nitrogen intermediates and reactive oxygen intermediates by macrophages and susceptibility to antimicrobial molecules. Therefore, it appears that there is no essential difference in virulence in terms of infectivity to human macrophages and lung cells between Mav strains isolated from NB-MAC disease and those from CA-MAC disease. These findings indicate the importance of further studies to elucidate the mechanism for the establishment of NB-type MAC diseases based on host immunological conditions rather than the pathogenic nature of MAC organisms themselves.
Asunto(s)
Complejo Mycobacterium avium/aislamiento & purificación , Complejo Mycobacterium avium/patogenicidad , Infección por Mycobacterium avium-intracellulare/microbiología , Infección por Mycobacterium avium-intracellulare/patología , Antiinfecciosos/farmacología , Bronquiectasia/microbiología , Bronquiectasia/patología , Línea Celular , Células Epiteliales/microbiología , Femenino , Humanos , Absceso Pulmonar/microbiología , Absceso Pulmonar/patología , Macrófagos/microbiología , Masculino , Pruebas de Sensibilidad Microbiana , Complejo Mycobacterium avium/crecimiento & desarrollo , Complejo Mycobacterium avium/inmunología , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , VirulenciaRESUMEN
Descending necrotizing mediastinitis (DNM) originating from deep cervical infection is a rare and serious clinical condition with a high mortality rate. Clinical feature of 5 patients undergone surgical drainage for DNM, between 2006 and 2009 were assessed. There were 3 male and 2 female patients whose age ranged from 57 to 83 years old (mean 69.8). All 5 patients had no underlying disease except for 1 patient with severe dental caries. The primary infections of these patients were tonsillitis and pharyngitis. The mean duration from onset of symptom to the referral to our hospital was 14 days (ranged 2 to approximately 41). Two patients underwent cervical drainage for upper mediastinum, and 3 patients were required mediastinal drainage by thoracotomy. There was no post-operative death. Early and aggressive surgical drainage of the neck and mediastinum by a multidisciplinary team of surgeons is very important in the treatment of DNM.
Asunto(s)
Mediastinitis/cirugía , Anciano , Anciano de 80 o más Años , Drenaje , Femenino , Humanos , Masculino , Mediastinitis/etiología , Persona de Mediana Edad , Faringitis/complicaciones , Tonsilitis/complicacionesRESUMEN
The mechanisms underlying cellular drug resistance have been extensively studied, but little is known about its regulation. We have previously reported that activating transcription factor 4 (ATF4) is upregulated in cisplatin-resistant cells and plays a role in cisplatin resistance. Here, we find out a novel relationship between the circadian transcription factor Clock and drug resistance. Clock drives the periodical expression of many genes that regulate hormone release, cell division, sleep-awake cycle and tumor growth. We demonstrate that ATF4 is a direct target of Clock, and that Clock is overexpressed in cisplatin-resistant cells. Furthermore, Clock expression significantly correlates with cisplatin sensitivity, and that the downregulation of either Clock or ATF4 confers sensitivity of A549 cells to cisplatin and etoposide. Notably, ATF4-overexpressing cells show multidrug resistance and marked elevation of intracellular glutathione. The microarray study reveals that genes for glutathione metabolism are generally downregulated by the knockdown of ATF4 expression. These results suggest that the Clock and ATF4 transcription system might play an important role in multidrug resistance through glutathione-dependent redox system, and also indicate that physiological potentials of Clock-controlled redox system might be important to better understand the oxidative stress-associated disorders including cancer and systemic chronotherapy.
Asunto(s)
Factor de Transcripción Activador 4/genética , Resistencia a Antineoplásicos/genética , Transactivadores/genética , Transcripción Genética , Factor de Transcripción Activador 4/metabolismo , Antineoplásicos/farmacología , Northern Blotting , Western Blotting , Proteínas CLOCK , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Inmunoprecipitación de Cromatina , Cisplatino/farmacología , Etopósido/farmacología , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Glutatión/metabolismo , Humanos , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patología , Oxidación-Reducción , Interferencia de ARN , Transactivadores/metabolismoRESUMEN
Acute lung injury (ALI) is one of most serious complications to occur after an esophagectomy for esophageal cancer. However, the pathogenesis of ALI is still unclear. The cytokine levels of pleural drainage fluid as well as peripheral blood were measured in 27 patients who had undergone an extended radical esophagectomy. Both the clinical factors and cytokine levels were compared between 11 patients with (group I) and 16 without ALI (group II). ALI occurred more frequently in patients who underwent colon interposition than in those who received a gastric tube reconstruction (86%vs 25%, P = 0.009). The operation time of group I was significantly longer than that of group II. A logistic regression analysis revealed colon interposition to be an independent factor associated with the ALI (P < 0.05). Postoperative anastomotic leakage and systemic inflammatory response syndrome (SIRS) occurred more frequently in group I than in group II (P < 0.01). Both the serum interleukin-6 (IL-6) and IL-8 levels of group I were significantly higher than those of group II. IL-1beta and tumor necrosis factor-alpha were undetectable in the peripheral blood, whereas they were detectable in the pleural effusion. The IL-1beta of pleural effusion was higher in group I than group II. In conclusion, greater surgical stress, such as a longer operative time, is thus considered to be associated with the first attack of ALI. The adverse events developing in the extra-thoracic site, such as necrosis and local infection around anastomosis may therefore be the second attack. Furthermore, ALI may cause not only SIRS but also other complications such as anastomotic leakage.
Asunto(s)
Neoplasias Esofágicas/cirugía , Esofagectomía , Complicaciones Posoperatorias , Síndrome de Dificultad Respiratoria/etiología , Anciano , Anastomosis Quirúrgica/efectos adversos , Carcinoma de Células Escamosas/cirugía , Colon/trasplante , Citocinas/análisis , Esofagectomía/métodos , Esofagoplastia , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Derrame Pleural/química , Respiración Artificial , Síndrome de Respuesta Inflamatoria Sistémica/etiología , Factores de TiempoRESUMEN
Somatically acquired mutations in the epidermal growth factor receptor (EGFR) gene in lung cancer are associated with significant clinical responses to gefitinib, a tyrosine kinase inhibitor (TKI) that targets EGFR. In our previous report, 42.2% of adenocarcinoma patients has EGFR mutations, and these mutations were more frequently found in women than in men, in well differentiated tumors than poorly differentiated tumors, and in patients who were never smokers than in patients who were current/former smokers. Retrospectively, we screened the EGFR gene of tumors in 37 NSCLC patients who had been treated with gefitinib. EGFR mutations were found in 22 patients. Gefitinib was effective (CR/PR) in 15 of 22 (68.2%) patients with mutations compared with none of 15 patients without mutations. Patients with EGFR mutations survived for a longer period than without the mutations after initiation of gefitinib treatment (p = 0.0005). Gefitinib was not effective in 3 patients with K-ras mutations. Three of 4 tumors obtained from patients with acquired resistant to gefitinib, had a secondary T790M mutation. No T790M mutation was detected in pretreatment tumors. Molecular targeted therapy using TKI indicates an effective therapy specifically in lung cancer patients with EGFR mutations, and analyses of mechanisms of resistance to TKI are necessary for establishment of tailor-made therapy.
Asunto(s)
Antineoplásicos/administración & dosificación , Genes erbB-1 , Neoplasias Pulmonares/tratamiento farmacológico , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Quinazolinas/administración & dosificación , Sistemas de Liberación de Medicamentos , Femenino , Gefitinib , Humanos , Masculino , Estudios RetrospectivosRESUMEN
Few reports on recurrence after thoracoscopic bullectomy for spontaneous pneumothorax specify the follow-up period and follow-up ratio. Because of the variation in follow-up periods, many reported recurrence rates were not comparable. Some reports compared simple recurrence rate (number of recurrent cases/number of operated cases) of different groups with different follow-up periods. In this study, we employ the Kaplan-Meier method along with a set of optimal follow-up periods and ratios in order to determine a more reliable recurrence rate. Consecutive 68 patients (74 surgical procedures) underwent thoracoscopic bullectomy for spontaneous pneumothorax at our institution between November 2000 and December 2005. A follow-up survey was conducted by phone to determine the rate of recurrent pneumothorax. The follow-up ratio and the mean follow-up period were 92.6% and 1,316 +/- 481 days, respectively. Postoperative recurrence was confirmed for 4 patients. The interval up to recurrence was 144, 345, 476 and 616 days after the bullectomy, respectively. All cases of recurrent pneumothorax occurred within 2 years following the bullectomy. The 1-year, 2-year and 3-year cumulative recurrence rate was 3.0%, 6.3% and 6.3%, respectively. In light of these findings, we feel that comparison analysis of pneumothorax recurrence rates should be evaluated using the Kaplan-Meier method, furthermore, our data suggests that a follow-up period of 2 or more years is advisable.
Asunto(s)
Neumotórax/cirugía , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias , Recurrencia , Adulto JovenRESUMEN
Tyrosinase is a rate-limiting enzyme in melanin biosynthesis and is specifically expressed in differentiated melanocytes. We have identified the enhancer element in the 5'-flanking region of the human tyrosinase gene that is responsible for its pigment cell-specific transcription and have termed it tyrosinase distal element (TDE) (positions -1861 to -1842). Transient expression assays showed that TDE confers efficient expression of a firefly luciferase reporter gene linked to the tyrosinase gene promoter in MeWo pigmented melanoma cells but not in HeLa cells, which do not express tyrosinase. TDE was specifically bound by nuclear proteins of MeWo and HeLa cells, the binding properties of which were indistinguishable in gel mobility shift assays. TDE contains the CATGTG motif in its center, and mutation analysis indicates that the CA dinucleotides of this motif are crucial for protein binding and pigment cell-specific enhancer function. The CATGTG motif is consistent with the consensus sequence recognized by a large family of transcription factors with a basic helix-loop-helix structure, which prompted us to examine the possible involvement of a ubiquitous transcription factor, USF, and a novel factor, microphthalmia-associated transcription factor (MITF), recently cloned as the human homolog of the mouse microphthalmia (mi) gene product. The mi phenotype is associated with a mutant mi locus and characterized by small eyes and loss of melanin pigments. Both USF and MITF are predicted to contain a basic helix-loop-helix structure and a leucine zipper structure. We provide evidence that USF binds to TDE, whereas we were unable to detect the DNA-binding activity of MITF. Transient coexpression assays showed that MITF specifically transactivates the promoter activity of the tyrosinase gene through the CATGTG motif of TDE but not the promoter of the ubiquitously expressed heme oxygenase gene, while USF is able to activate both promoters. These results indicate that MITF is a cell-type-specific factor that is capable of activating transcription of the tyrosinase gene.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Melanocitos/metabolismo , Monofenol Monooxigenasa/genética , Factores de Transcripción/fisiología , Secuencia de Bases , Sitios de Unión , Clonación Molecular , Proteínas de Unión al ADN/metabolismo , Elementos de Facilitación Genéticos , Regulación Enzimológica de la Expresión Génica , Secuencias Hélice-Asa-Hélice , Humanos , Técnicas In Vitro , Leucina Zippers , Factor de Transcripción Asociado a Microftalmía , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Regiones Promotoras Genéticas , Unión Proteica , ARN Mensajero/genética , Transcripción Genética , Células Tumorales Cultivadas , Factores Estimuladores hacia 5'RESUMEN
A novel cis-acting regulatory element (designated BTE for basic transcription element) was found in the region proximal to the TATA sequence of the P-450c gene by the use of deletion mutations. This DNA element is considered to be involved in the basic transcription of the gene and does not show distinct enhancer activity in itself. Together with the XRE sequence (A. Fujisawa-Sehara, K. Sogawa, M. Yamane, and Y. Fujii-Kuriyama, Nucleic Acids Res. 15:4179-4191, 1987), however, this sequence is required for a high inducible expression of the P-450c gene in response to xenobiotic inducers. The BTE sequence contained the GC box consensus sequence and half of the NF-1-binding consensus or CAT box sequence, but their synthetic oligonucleotides, used as competitors in the gel mobility shift assays, did not compete with the BTE sequence for the binding protein, suggesting that the BTE sequence functions as a different recognition sequence from that for Sp1 or NF-1. Analogous sequences to BTE are found in the region proximal to the TATA sequence of other genes, especially other P-450 genes with different modes of regulation, suggesting that the BTE sequence plays a common regulatory role in basic transcription of genes including a group of the P-450 superfamily. The ubiquitous distribution of nuclear factor(s) binding to this element supports this suggestion.
Asunto(s)
Sistema Enzimático del Citocromo P-450/genética , Regulación de la Expresión Génica , Genes , Regiones Promotoras Genéticas , Animales , Secuencia de Bases , Línea Celular , Cloranfenicol O-Acetiltransferasa/genética , Deleción Cromosómica , Datos de Secuencia Molecular , Mutación , Sondas de Oligonucleótidos , Ratas , Homología de Secuencia de Ácido Nucleico , TransfecciónRESUMEN
C57BL/6 mice inoculated i.p. with 3LL tumor cells were treated by local combination therapy with Nocardia rubra cell wall skeleton (N-CWS) and recombinant interleukin 2 (rIL-2). The combination treatment significantly prolonged their survival and augmented lymphokine-activated killer (LAK) activity of peritoneal cavity lymphocytes (PCL), compared with treatments with rIL-2 alone or N-CWS alone. After in vitro culture of peritoneal exudate mononuclear cells with rIL-2, the nonadherent population derived from N-CWS-injected tumor-bearing mice showed a significantly higher LAK activity than did that population derived from saline solution-injected mice. When N-CWS-induced PCL were cocultured with either N-CWS-induced macrophages or control macrophages in the presence of rIL-2, their LAK activity was higher than that of control PCL. Therefore, it was suggested that N-CWS-induced PCL themselves have a more potent ability as precursors of LAK cells. Phenotypic analysis on PCL populations revealed that N-CWS-induced PCL contained increased proportions of CD3+CD4-CD8- cells and asialo GM-1+ cells compared with control PCL. These results suggest that N-CWS selectively accumulates potent LAK precursors, namely, CD3+CD4-CD8- T-cells and asialo GM-1+ natural killer cells, at the injection site and that LAK cells are efficiently induced by subsequent administration of rIL-2.
Asunto(s)
Toxinas Bacterianas/farmacología , Citoesqueleto/inmunología , Inmunoterapia , Interleucina-2/farmacología , Células Asesinas Activadas por Linfocinas/efectos de los fármacos , Neoplasias Pulmonares/terapia , Activación de Linfocitos/efectos de los fármacos , Melanoma/terapia , Nocardia , Animales , Antígenos de Diferenciación de Linfocitos T/biosíntesis , Complejo CD3 , Antígenos CD4/análisis , Antígenos CD8/análisis , Citotoxicidad Inmunológica/efectos de los fármacos , Modelos Animales de Enfermedad , Inmunofenotipificación , Células Asesinas Activadas por Linfocinas/inmunología , Neoplasias Pulmonares/inmunología , Masculino , Melanoma/inmunología , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Receptores de Antígenos de Linfocitos T/biosíntesis , Receptores de Interleucina-2/metabolismo , Proteínas Recombinantes/farmacologíaRESUMEN
Antitumor activities of pleural cavity macrophages (PCM) and pleural cavity lymphocytes (PCL) in lung cancer patients were examined. The effect of coculture supernatants of PCL and autologous tumor cells on the cytostatic activity of macrophages was also examined. Cytostatic activity of PCM was not affected by an advance of metastasis to regional lymph nodes or increase of tumor size and difference of histological type. However, the cytostatic activity of PCM was markedly augmented when pleural invasion was limited to within the visceral pleura although it was low when pleural invasion was absent or extended beyond the visceral pleura. On the other hand, PCL did not exert any cytolytic activity against various tumor target cells. However, coculture supernatants of PCL and autologous tumor cells exhibited the activity of macrophage-activating factor against guinea pig peritoneal macrophages. Furthermore, the higher the cytostatic activity of PCM, the higher the macrophage-activating factor activity of the coculture supernatant of PCL and autologous tumor cells was. These results suggested that antitumor activity of PCM was controlled by specifically sensitized PCL through lymphokines.
Asunto(s)
Neoplasias Pulmonares/patología , Linfocitos/inmunología , Macrófagos/inmunología , Pleura/citología , Animales , Citotoxicidad Inmunológica , Cobayas , Humanos , Metástasis Linfática , PronósticoRESUMEN
The purpose of this study is to clarify the significance of antitumor activity of alveolar macrophages (AM) in lung cancer patients. AM from tumor-bearing and non-tumor-bearing segments were obtained separately by the lavage of bronchoalveolar tracts of resected lungs of 74 patients with primary lung cancer. Cytostatic activity (CTS) of AM obtained from non-tumor-bearing segments was stable in spite of enlargement of tumor size or progression of N factor. In contrast, CTS of AM obtained from tumor-bearing segments may be augmented at Stage II as compared with Stage I, and suppressed with an advance of stage from II through IV, although the number of Stage II patients was as small as three. Moreover, CTS of AM from tumor-bearing segments was suppressed in N2 as compared with N0 or N1, and also it was suppressed as compared with that of AM from non-tumor-bearing segments in N2 disease. CTS of AM from smokers was suppressed as compared with that of nonsmokers in both tumor-bearing and non-tumor-bearing segments. These results suggest that lung cancer cells or their products may suppress antitumor activity of AM in the tumor-bearing segments at advanced stages, and cigarette smoking is a suppressive factor on antitumor activity of AM.
Asunto(s)
Citotoxicidad Inmunológica , Neoplasias Pulmonares/inmunología , Macrófagos/inmunología , Alveolos Pulmonares/inmunología , Femenino , Humanos , Masculino , Estadificación de Neoplasias , FumarRESUMEN
The percentage of tumor-associated macrophages recovered (TAMR) and antitumoral activity of tumor-associated macrophages (TAM) were examined in 77 patients with resectable primary lung cancer. TAM was obtained by plastic adherence following trypsinization. TAMR increased from Stage I to Stage II and decreased in Stage III. It also increased in N1 as compared with N0 and N2 but was unrelated to tumor size. However, the cytostatic activity of TAM declined with advance in stage of the disease and an increase of tumor size, but it was relatively unaffected by the presence of metastasis to regional lymph nodes. There was no correlation between TAMR and the recurrence rate; however, cytostatic activity of TAM was correlated significantly with the prognosis of totally resected cases. TAMR and cytostatic activity of TAM tended to be lower in palliatively resected cases. These results suggest that the assessment of the antitumor activity of TAM, but not merely TAMR, may give prognostic information for lung cancer patients.
Asunto(s)
Neoplasias Pulmonares/inmunología , Macrófagos/inmunología , Adenocarcinoma/inmunología , Carcinoma de Células Escamosas/inmunología , Femenino , Humanos , Neoplasias Pulmonares/cirugía , Metástasis Linfática , Masculino , Pronóstico , Timidina/metabolismo , TritioRESUMEN
High- (Cl-33H) and low- (Cl-35L) metastatic clones were established from a methylcholanthrene-induced rat fibrosarcoma (FMQ-100). The modal chromosome numbers of the two clones were different. These clones grew in in vitro culture, showing similar growth rate and saturation density. However, in in vivo experiments, Cl-33H exhibited a higher tumor growth rate, tumorigenicity, spontaneous metastatic potential, and experimental metastatic potential than did Cl-35L. Alveolar macrophages obtained from normal syngeneic rats stimulated growth of these clones in vitro, as assessed by [3H]thymidine uptake. Moreover, this effect was greater on Cl-33H than Cl-35L. The growth-promoting effect of macrophages was also observed under the in vitro condition of lack of direct contact between macrophages and tumor cells. These results suggested the possibility that alveolar macrophage-derived growth-promoting factors play some role in the development of pulmonary metastasis in this tumor system, and the difference of susceptibility to the growth-promoting factors might be one of the causes of the different metastatic potentials of Cl-33H and Cl-35L.
Asunto(s)
Fibrosarcoma/patología , Metástasis de la Neoplasia , Animales , Línea Celular , Sustancias de Crecimiento/fisiología , Cariotipificación , Neoplasias Pulmonares/secundario , Macrófagos/fisiología , Metilcolantreno , Alveolos Pulmonares/inmunología , Alveolos Pulmonares/patología , RatasRESUMEN
Previous studies demonstrated that injecting mice with the cytokine interleukin 12 (IL-12) could significantly suppress the growth of a number of tumors, including murine B16 melanoma. In this report, the persistence of the antitumor effects of IL-12 is investigated. The i.p. injection of IL-12 (0.1 microg) on days 14, 16, 18, 20, and 22 was found to significantly suppress the growth of s.c. inoculated B16 melanoma for up to 2 weeks after the last injection of IL-12. Interestingly, the IL-12 serum level 4 days after the last injection of IL-12 was significantly elevated in tumor-bearing mice compared with that of IL-12-treated normal mice. The in vivo depletion of either CD4+ or CD8+ T cells abrogated the antitumor activity of IL-12 and diminished the apparent autocrine stimulation of IL-12 release seen after IL-12 treatment. Resection of the tumor-draining lymph nodes (LNs) but not of the spleen abrogated the antitumor effect of IL-12 treatment as well as the elevation of serum IL-12. Expression of mRNA encoding IL-12 as well as CD40 ligand (CD40L) was detected in the tumor-draining LNs but not in the spleen of tumor-bearing mice after IL-12 treatment. Furthermore, the antitumor activity observed after IL-12 treatment was diminished by the in vivo administration of either anti-IL-12 or anti-CD40L monoclonal antibodies. Collectively, these results suggest that the endogenous production of IL-12 resulting from the CD40-CD40L interaction between antigen-presenting cells and CD4+ T cells in the tumor-draining LNs may play a role in the persistence of the antitumor effects seen after IL-12 treatment.
Asunto(s)
Inmunoterapia , Interleucina-12/administración & dosificación , Melanoma Experimental/terapia , Animales , Linfocitos T CD4-Positivos/inmunología , Antígenos CD40/inmunología , Antígenos CD40/metabolismo , Linfocitos T CD8-positivos/inmunología , Femenino , Interleucina-12/biosíntesis , Interleucina-12/sangre , Interleucina-12/inmunología , Melanoma Experimental/sangre , Melanoma Experimental/inmunología , Ratones , Ratones Endogámicos C57BL , Células Tumorales CultivadasRESUMEN
A randomly controlled study of chemotherapy versus chemoimmunotherapy was performed in patients with operable lung cancer from November 1977 to June 1981. The immunotherapy consisted of an intrapleural instillation of Nocardia rubra cell wall skeleton (N-CWS) followed by serial intradermal N-CWS. A total of 119 patients were entered into this trial. There were 64 evaluable patients in the control group and 52 evaluable patients in the N-CWS group. N-CWS treatment was effective in terms of prolongation of duration of remission for all operable patients. Although significant improvement in the survival rate was not observed in patients at Stages I and II (p less than 0.10), it was observed in the curative operation group (p less than 0.05). The mode of recurrence was classified as local recurrence and distant metastasis in the curative operation group. The rates of distant metastasis were 34.0 and 18.9%, respectively, in the control and the N-CWS groups. The rate of local recurrence was 14.9% in the control group; however, no local recurrence was observed in the N-CWS group. These results indicate the clinical effectiveness of the N-CWS treatment, especially in curatively resectable lung cancer. No serious side effect was observed during this trial.
Asunto(s)
Inmunoterapia/métodos , Neoplasias Pulmonares/terapia , Mucoproteínas/inmunología , Nocardia/inmunología , Adulto , Anciano , Esqueleto de la Pared Celular , Ensayos Clínicos como Asunto , Terapia Combinada , Femenino , Humanos , Neoplasias Pulmonares/mortalidad , Neoplasias Pulmonares/cirugía , Masculino , Persona de Mediana Edad , Mucoproteínas/efectos adversos , Ácidos Micólicos/efectos adversos , Recurrencia Local de Neoplasia , Distribución AleatoriaRESUMEN
Intrapleural instillations of recombinant interleukin 2 (RIL-2) were performed in 11 patients with malignant pleurisy due to lung cancer. Kinetic studies on RIL-2 concentration in the pleural effusion and serum revealed relatively long-term maintenance of detectable levels of RIL-2 (over 24 h in the pleural effusion and over 8 h in the serum). Clinically, pleural effusions and cancer cells in the effusions disappeared in 9 of the 11 patients 4 to 10 days after the start of the treatment. Lymphokine-activated killer cells were induced in the effusions of responders who exhibited the disappearance of pleural effusion and cancer cells from the effusion, but not in those of the nonresponders. This induction of lymphokine-activated killer cells may result in the disappearance of cancer cells and pleural effusions. Cytological examination of pleural effusions revealed increases of lymphoblasts, immunoblastic large lymphocytes, and eosinophiles in number and proportion in the responder, although such a phenomenon could not be observed in the nonresponders. Main and frequent side effects of intrapleural instillations of RIL-2 were fever up to 39 degrees C, transient increase of pleural effusion, and eosinophilia. No serious side effect was encountered in our experience.
Asunto(s)
Interleucina-2/uso terapéutico , Células Asesinas Naturales/inmunología , Neoplasias Pulmonares/terapia , Linfocinas/farmacología , Pleuresia/terapia , Adulto , Anciano , Citotoxicidad Inmunológica , Femenino , Humanos , Interleucina-2/efectos adversos , Interleucina-2/análisis , Neoplasias Pulmonares/inmunología , Masculino , Persona de Mediana Edad , Derrame Pleural/metabolismo , Pleuresia/inmunología , Proteínas Recombinantes/uso terapéuticoRESUMEN
Adhesion molecules are involved in intracellular signaling in various physiological and pathological processes including metastasis and growth of tumor cells. Tumor cells interact with various host cells as well as with extracellular matrices through certain adhesion molecules such as integrins. We here propose that stimulation of beta1 integrin reduces intercellular adhesion molecule (ICAM)-1-mediated interaction of lung cancer cells with CTLs. This concept is based on the following findings: (a) engagement of beta1 integrins on certain lung cancer cells by a specific antibody or by ligand matrices such as fibronectin and collagen markedly reduced ICAM-1 expression on the cell surface and induced sICAM-1; (b) down-regulation of ICAM-1 by stimulation of beta1 integrins was abrogated by tyrosine kinase inhibitors or by transfection of dominant negative truncations of focal adhesion kinase (FAK); (c) engagement of beta1 integrins also reduced ICAM-1-dependent adhesion of lung cancer cells to T cells, a process completely inhibited by tyrosine kinase inhibitors and by transfection of dominant negative forms of FAK; and (d) stimulation of beta1 integrins prevented killing of lung cancer cells by autologous CTLs. In malignant tumors, cancer cells, including lung cancer cells, are surrounded by extracellular matrix proteins such as fibronectin and collagen. This suggests that the engagement of beta1 integrins by matrix proteins potentially occurs in cancer cells in vivo and that continuous stimulation via beta1 integrins reduces ICAM-1-expression, ICAM-1-mediated adhesion of cancer cells to CTLs and their killing by CTLs. Our results suggest that such processes can lead to the escape of lung cancer cells in vivo from immunological surveillance.
Asunto(s)
Integrina beta1/fisiología , Molécula 1 de Adhesión Intercelular/biosíntesis , Neoplasias Pulmonares/patología , Proteínas Tirosina Quinasas/fisiología , Anticuerpos Monoclonales/farmacología , Adhesión Celular/fisiología , Colágeno/metabolismo , Colágeno/farmacología , Citotoxicidad Inmunológica , Regulación hacia Abajo , Fibronectinas/metabolismo , Fibronectinas/farmacología , Quinasa 1 de Adhesión Focal , Proteína-Tirosina Quinasas de Adhesión Focal , Regulación Neoplásica de la Expresión Génica , Humanos , Integrina beta1/biosíntesis , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/fisiología , Neoplasias Pulmonares/metabolismo , Antígeno-1 Asociado a Función de Linfocito/fisiología , Fosforilación , Proteínas Tirosina Quinasas/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Linfocitos T/patología , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/patologíaRESUMEN
Tumor-reactive T cells, known as tumor-infiltrating lymphocyte(TIL)s are known to infiltrate various tumors. Although TILs exert cytotoxic activities against tumor cells, only a small percentage of tumors usually contain TILs that specifically react to tumor antigens. Because the exact role of these lymphocytes is unclear, we investigated the mechanisms of migration and adhesion of TILs to bone metastatic tumors, particularly to osteoblasts and bone marrow-derived stromal cell(BMSC)s. Histopathological examination showed that most TILs in secondary bone metastatic tumors (from primary tumors in the lung or breast) were found in the supporting tissue stroma between the bone and tumor mass. Cultured TILs (obtained from breast tumors) adhered spontaneously to osteoblasts and BMSCs (obtained from patients with osteoarthritis) without exogenous stimulation. Adhesion was further enhanced by chemokines macrophage inflammatory protein (MIP)-1alpha and MIP-1beta. TILs highly expressed activation antigens CD25 and CD69. A spontaneous activation of an integrin, lymphocyte function-associated antigen-1 (LFA-1), was also detected on TILs. TILs produced high concentrations of MIP-1alpha and MIP-1beta and spontaneous polymerization of cytoskeletal F-actin was observed in these cells. Adhesion of TILs to osteoblasts and BMSCs via LFA-1 and very late antigen-4 was associated with the production of osteoclastogen interleukin 6 by the latter cells. Our results indicate that integrins on TILs are activated in an autocrine manner by MIP-1alpha and MIP-1beta, and that treatment with the chemokines increases the binding of TILs on osteoblasts and stromal cells via a mechanism involving intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 as targets for the integrin. Our data also indicated that interactions between TILs and osteoblasts/stromal cells lead to the secretion by the latter of the osteoclastogenic cytokine interleukin 6.