RESUMEN
Antiphospholipid syndrome is characterized by multiple arterial and/or venous thrombotic events, recurrent fetal losses in the presence of antiphospholipid antibodies (aPL). Catastrophic antiphospholipid syndrome is a life-threatening, rare subset of antiphospholipid syndrome when the thrombotic events affect at least three organs, and clinical manifestations develop simultaneously or within a week. Diagnostically, small vessel occlusions can be detected by histopathology in the presence of aPL. Our case report describes an 18-year-old man who has been treated for antiphospholipid syndrome associated with systemic lupus erythematosus (SLE) since 2011. The clinical findings were dominated by recurrent deep vein thrombosis, and severe proteinuria caused by lupus nephritis, accompanied by mild serological and laboratory findings. The patient was hospitalized in March 2014 because of severe thrombocytopenia and infective diarrhoea. At this time the renal functions deteriorated rapidly. Simultaneously, left upper extremity paresis was observed; computed tomography showed ischaemic lesions in the territory of the middle cerebral artery. Abdominal discomfort and pain occurred. On computed tomography scan ischaemic lesions were seen in the spleen, the right kidney and the coeliac trunk. Laboratory and serological findings verified the presence of aPL and anti-DNA antibodies, anaemia and thrombocytopenia. Based on the above-mentioned clinical and laboratory findings, the diagnosis of catastrophic antiphospholipid syndrome was established. Anticoagulation, corticosteroids and plasma exchange treatment, as well as haemodiafiltration were initiated. Although the thrombotic cascade decelerated following these interventions, we could not see an improvement in the renal function. Rituximab treatment was started, leading to a significant improvement in renal function. After 5 weeks of treatment the patient was discharged from hospital.
Asunto(s)
Síndrome Antifosfolípido/complicaciones , Factores Inmunológicos/uso terapéutico , Nefritis Lúpica/complicaciones , Rituximab/uso terapéutico , Trombosis/inmunología , Síndrome Antifosfolípido/tratamiento farmacológico , Síndrome Antifosfolípido/patología , Humanos , Riñón/ultraestructura , Nefritis Lúpica/tratamiento farmacológico , Nefritis Lúpica/patología , Masculino , Trombosis/tratamiento farmacológico , Trombosis/patología , Adulto JovenRESUMEN
Little is known about the altered lipid metabolism-related transcriptional events occuring in sebaceous glands of patients with acne vulgaris. Peroxisome proliferator-activated receptor (PPAR)γ, a lipid-activated transcription factor, is implicated in differentiation and lipid metabolism of sebocytes. We have observed that PPARγ and its target genes, ADRP (adipose differentiation related protein) and PGAR (PPARγ angioprotein related protein) are expressed at lower levels in sebocytes from patients with acne than in those from healthy controls (HCs) Furthermore, endogenous PPARγ activator lipids such as arachidonic acid-derived keto-metabolites (e.g. 5KETE, 12KETE) are increased in acne-involved and nonacne-involved skin of patients with acne, compared with skin from healthy individuals. Our findings highlight the possible anti-inflammatory role of endogenous ligand-activated PPARγ signaling in human sebocyte biology, and suggest that modulating PPARγ- expression and thereby signaling might be a promising strategy for the clinical management of acne vulgaris.
Asunto(s)
Acné Vulgar/metabolismo , PPAR gamma/metabolismo , Glándulas Sebáceas/metabolismo , Transducción de Señal/fisiología , Adulto , Análisis de Varianza , Proteína 4 Similar a la Angiopoyetina , Angiopoyetinas/metabolismo , Estudios de Casos y Controles , Eicosanoides/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Inmunohistoquímica , Masculino , Perilipina-2/metabolismo , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Factor XIII subunit A (FXIII-A) is used as a diagnostic marker in a wide range of dermatological diseases ranging from inflammatory lesions to malignancies, although neither the cell types responsible for its expression nor the mechanism(s) resulting in its local accumulation in pathological conditions have been characterized. OBJECTIVE: In this study, we aimed to gain information on the cells showing an immunohistochemical reaction for FXIII-A and answer the question whether macrophages and/or dendritic cells are labelled for FXIII-A. METHODS: We carried out our studies on samples of granuloma annulare (GA) and necrobiosis lipoidica (NL), the prime examples for granulomatous skin lesions with a non-infectious background in which extracellular matrix remodelling is a key feature without any sign of malignant transformation. We used markers for macrophages and dendritic cells in combination with the detection of FXIII-A in double labelling immunohistochemical reactions. RESULTS: We demonstrated that FXIII-A positivity clearly distinguishes macrophages (CD163+/FXIII-A+) from dendritic cells (CD11c+/FXIII-A-) not only in the normal dermis as previously described by Zaba et al. (J Clin Invest 2007; 117: 2517-2525) but also in the pathological conditions of GA and NL. Detecting the expression of DC-SIGN/CD209 and mannose receptor molecules on FXIII-A+ macrophages we confirmed that FXIII-A is expressed in the alternatively activated macrophages. However, while DC-SIGN/CD209 was invariably expressed on FXIII-A+ cells both in normal and pathological conditions of GA/NL (98.7% vs. 93.5/96%), mannose receptor was only partially coexpressed with FXIII-A (94.8% vs. 74.7/52.2%), suggesting that FXIII-A+ macrophages do not represent a homogenous population. CONCLUSIONS: FXIII-A selectively marks macrophages and distinguishes them from dendritic cells. The presence of FXIII-A is not a disease-specific marker but indicates a possible common mechanism of macrophage activation in various dermatological diseases.
Asunto(s)
Células Dendríticas/clasificación , Factor XIIIa/análisis , Granuloma Anular/inmunología , Macrófagos/clasificación , Técnica del Anticuerpo Fluorescente , HumanosRESUMEN
BACKGROUND: The formation of metastases and the efficacy of systemic therapies in cutaneous malignant melanoma (CMM) depend on the characteristics of the tumour cells and the host immune response. Aberrant expression of metallothionein (MT) has been observed in several types of cancers with poor prognoses. OBJECTIVE: To perform an immunohistochemical study on primary CMM comparing the MT expression of tumours without metastases (n = 23) to that of samples with haematogenous metastases (n = 23) and to examine the correlation between MT staining and immunological markers relevant in CMM progression. METHODS: The immunohistochemical labelling of different tumour sections was analysed using tissue microarrays for the evaluation of the suitability of this method in future studies. RESULTS: Our results suggest that MT overexpression is significantly more frequent in primary CMM with haematogenous metastases (P = 0.018) and that the overexpression is independent of the Breslow tumour thickness (R = 0.102, P = 0.501). Interestingly, MT overexpression of the tumour cells was correlated with the presence of tumour-infiltrating CD68(+) macrophages (P = 0.003), a known predictive factor for melanoma progression, thereby suggesting a role for MT in the development of a defective host immune response. Furthermore, the presence of CD163(+) macrophages infiltrating the tumours correlated with metastasis formation (P < 0.001), whereas the presence CD1a(+) dendritic cells surrounding the tumours was associated with a lower risk of haematogenous spread (P = 0.003). CONCLUSION: Our results demonstrate that MT may represent a suitable prognostic factor that can characterize the metastasising ability of CMM and the tumour-promoting host immune response.
Asunto(s)
Macrófagos/patología , Melanoma/metabolismo , Metalotioneína/metabolismo , Neoplasias Cutáneas/metabolismo , Antígenos CD/inmunología , Progresión de la Enfermedad , Femenino , Humanos , Macrófagos/inmunología , Masculino , Melanoma/inmunología , Melanoma/patología , Metástasis de la Neoplasia , Factores de Riesgo , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patologíaRESUMEN
The aim of the present study was to assess the autoantibody profile, dominant clinical symptoms and cluster characteristics of different mixed connective tissue disease (MCTD phenotypes. Two-hundred-and-one patients with MCTD were followed-up longitudinally. Five clinical parameters, Raynaud's phenomenon, pulmonary artery hypertension (PAH), myositis, interstitial lung disease (ILD), erosive arthritis and five auto-antibodies besides anti-U1RNP, antiendothelial cell antibodies (AECA), anti-CCP, anti-cardiolipin (anti-CL), anti-SSA/SSB and IgM rheumatoid factor (RF) were selected for cluster analysis. The mean age of patients was 52.9 ± 12.4 years and the mean follow-up of the disease was 12.5 ± 7.2 years. Patients were classified into three cluster groups. Cluster 1 with 77 patients, cluster 2 with 79 patients and cluster 3 with 45 patients. In cluster 1 the prevalence of PAH (55.8%; p < 0.001), Raynaud's phenomenon (92.2%; p < 0.001) and livedo reticularis (24.6%, p < 0.001) was significantly greater than in cluster 2 and 3. In cluster 2, the incidence of ILD (98.7%; p < 0.001), myositis (77.2%; p < 0.001), and esophageal dysmotility (89.8%; p < 0.001) was significantly greater than that in cluster 1 and 3. In cluster 3, anti-CCP antibodies were present in 31 of 45 patients (68.8%) with erosions. Anti-CCP antibodies were present in 37 of 42 patients (88.0%) with erosions. PAH, angina, venous thrombosis was observed in cluster 1 and pulmonary fibrosis in cluster 2, musculosceletal damage, gastrointestinal symptoms and osteoporotic fractures were most frequent in cluster 3. Cumulative survival assessment indicated cluster 1 patients having the worst prognosis. Cluster analysis is valuable to differentiate among various subsets of MCTD and useful prognostic factor regarding the disease course.
Asunto(s)
Enfermedad Mixta del Tejido Conjuntivo/epidemiología , Adulto , Anciano , Análisis de Varianza , Artritis/epidemiología , Autoanticuerpos/sangre , Biomarcadores/sangre , Distribución de Chi-Cuadrado , Análisis por Conglomerados , Progresión de la Enfermedad , Hipertensión Pulmonar Primaria Familiar , Femenino , Humanos , Hungría/epidemiología , Hipertensión Pulmonar/epidemiología , Incidencia , Estudios Longitudinales , Enfermedades Pulmonares Intersticiales/epidemiología , Masculino , Persona de Mediana Edad , Enfermedad Mixta del Tejido Conjuntivo/clasificación , Enfermedad Mixta del Tejido Conjuntivo/diagnóstico , Enfermedad Mixta del Tejido Conjuntivo/inmunología , Enfermedad Mixta del Tejido Conjuntivo/mortalidad , Miositis/epidemiología , Fenotipo , Prevalencia , Pronóstico , Enfermedad de Raynaud/epidemiología , Análisis de Supervivencia , Factores de TiempoRESUMEN
We describe a rare case of concurrent polymyositis and Crohn's disease in a female patient. A 69-year-old female presented in December 2007 with a 5-month history of proximal muscle weakness, pain, fatigue and difficulty in walking and swallowing. Blood tests revealed elevated creatine kinase (3,429 U/l) and lactate dehydrogenase (2,013 U/l) levels. Magnetic resonance imaging found lumbar disc protrusion. Review by immunologists showed a diagnosis of idiopathic inflammatory myopathy. Though electromyography and muscle biopsy at this point were non-specific, corticosteroid treatment was commenced. Her condition worsened precipitously leading to hospitalisation under immunologists. As the provisional diagnosis was polymyositis, we commenced 1.5 mg/kg per day corticosteroid but her muscle power did not improve. Recurrent abdominal symptoms lead to ultrasonography showing intestinal inflammation. While tumour markers were elevated, thorough investigation failed to identify a tumour. Corticosteroid therapy was continued. Persistent abdominal symptoms lead to repeat colonoscopy and biopsy confirming Crohn's disease. Repeat electromyography and muscle biopsy confirmed the diagnosis of polymyositis. Her corticosteroids were tapered off and 5-aminosalicylic acid and azathioprine were started. Her myositic symptoms gradually abated with improvement in her Crohn's disease. She is now able to walk independently and takes 8 mg/day corticosteroids and her muscle enzyme levels are normal. Remember rare systemic associations when dealing with immune-mediated disease. Consider myositis in the differential diagnosis of Crohn's disease associated myopathy. Treating Crohn's disease may lead to improvement in steroid-resistant myositis where the two are associated.
Asunto(s)
Enfermedad de Crohn/complicaciones , Polimiositis/complicaciones , Anciano , Antiinflamatorios/uso terapéutico , Antiinflamatorios no Esteroideos/uso terapéutico , Azatioprina/uso terapéutico , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/tratamiento farmacológico , Femenino , Glucocorticoides/uso terapéutico , Humanos , Inmunosupresores/uso terapéutico , Desplazamiento del Disco Intervertebral/patología , Vértebras Lumbares/patología , Imagen por Resonancia Magnética , Mesalamina/uso terapéutico , Polimiositis/diagnóstico , Polimiositis/tratamiento farmacológico , Resultado del TratamientoRESUMEN
BACKGROUND: Extravascular activation of the coagulation system and consequent fibrin deposition is involved in the pathomechanism of chronic bronchoalveolar inflammatory diseases. The turnover of extravascular fibrin is attenuated by its cross-linking with activated factor XIII (FXIII). OBJECTIVES: Determination of cellular and plasmatic forms of FXIII and their correlation with D-dimer level in the bronchoalveolar lavage fluid (BALF) from healthy children and from children with bronchoalveolar inflammation. PATIENTS AND METHODS: Highly sensitive immunoassays were used for the quantitation of cellular and plasma FXIII and D-dimer in the BALF of children with recurrent wheezy bronchitis and fibrosing alveolitis. BALF was investigated for FXIII-containing cells by flow cytometry. RESULTS AND CONCLUSIONS: In the BALF of controls a low amount of the cellular form of FXIII (FXIII A2) and D-dimer were measured, while plasma FXIII (FXIII A2B2) was absent. Alveolar macrophages represented the single cell population in BALF that contained FXIII. In the BALF of both patients' groups the concentration and the total amount of FXIII A2 was significantly elevated, and plasma FXIII also appeared in the BALF of most patients. The D-dimer concentration was also elevated in the patients' groups and it correlated both with plasma FXIII and neutrophil count. These findings suggest that FXIII A2 is released from activated or injured alveolar macrophages into the bronchoalveolar lining fluid and in bronchoalveolar inflammatory diseases, FXIII A2B2 also leaks out from the capillaries. By cross-linking fibrin and inhibitors of fibrinolysis to fibrin, FXIII might be a key regulator of fibrin turnover in the extravascular compartment.
Asunto(s)
Bronquios/patología , Factor XIII/metabolismo , Inflamación/patología , Alveolos Pulmonares/patología , Adolescente , Bronquitis/patología , Líquido del Lavado Bronquioalveolar , Capilares/patología , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Factor XIII/biosíntesis , Deficiencia del Factor XIII/diagnóstico , Factor XIIIa/biosíntesis , Femenino , Fibrina/metabolismo , Productos de Degradación de Fibrina-Fibrinógeno/biosíntesis , Fibrinólisis , Citometría de Flujo , Humanos , Lactante , Macrófagos/metabolismo , Masculino , Neutrófilos/metabolismo , Factores de TiempoRESUMEN
Twenty temporal bones (TBs) were removed from autopsy cases and prepared for immunohistochemical examination. Ten TBs were free of ear disease whereas the other ten TBs showed the signs of chronic otitis media. Expression of markers for monocyte-macrophages (25F9, 27E10) and natural killer cells (anti-Leu-11) was examined immunohistochemically. There were no specific positive stainings with 25F9 or anti-Leu-11 antibodies in any of the specimens. Staining for 27E10 was found to be negative in each section obtained from normal cochlea. However, 27E10 positivity was detected in three of ten TBs with signs of chronic ear inflammation. This positivity can be explained by two theories: (1) activated monocytes can enter the inner ear from the systemic circulation as a consequence of chronic antigen challenge; (2) mesothelial cells could become activated as a result of a cross-reaction, with resultant positivity. Development of sensorineural hearing loss in some cases of chronic otitis media may be due to these immunological reactions.
Asunto(s)
Biomarcadores/análisis , Pérdida Auditiva Sensorineural/inmunología , Otitis Media/inmunología , Adulto , Anciano , Cadáver , Enfermedad Crónica , Técnicas de Cultivo , Oído Interno/inmunología , Oído Interno/patología , Pérdida Auditiva Sensorineural/patología , Humanos , Inmunohistoquímica , Células Asesinas Naturales/inmunología , Persona de Mediana Edad , Monocitos , Otitis Media/patología , Valores de Referencia , Sensibilidad y Especificidad , Hueso TemporalRESUMEN
The aim of this study was to obtain baseline data on the recently described special form of single cell death, apoptosis, in normal human inner ears. For this purpose, in situ end-labeling of the fragmented DNA was applied, in conjunction with apoptosis-related markers, to detect cellular elements showing programmed cell death in decalcified and paraffin-embedded tissues. Over 20 specimens were analyzed which were obtained from autopsy cases with no history of acoustic lesions confirmed by histopathology. Based on staining results, we saw no apoptotic signs in the majority of normal adult inner ears. An apoptotic cell captured in the Reissner's membrane of the cochlea from an old patient may, however, indicate an age-related subtle cell loss with the process of apoptosis. Nevertheless, the fact that more apoptosis was not found in our cases suggests that this phenomenon does not contribute significantly to the tissue homeostasis in the adult inner ear under normal conditions. These data are in accordance with our immunohistochemical findings on the p53 nucleoprotein, and proliferating cell nuclear antigen expression since there was no staining in any of the cellular elements, including the mesenchymal cells. This reflects a stationary and stable condition of cells of the vestibular and the cochlear structures, probably to maintain their integrity and the fine sensory functions. As opposed to the above findings, during inner ear development, the epithelial cells lining the cochlear lumen, the ossifying cartilage of the temporal bone, and the mesenchymal cells show different degrees of proliferation in combination with single cell death as signs of maturation of the vestibular and the cochlear apparatus. In addition, apoptosis has been demonstrated in cells of the cochlear stria vascularis from an adult patient treated with high doses of cisplatin, vinblastine and bleomycin prior to death. Furthermore, a wide range of apoptosis could be induced experimentally in a normal ear by an external perfusion of actinomycin D (ActD), which is known to produce programmed cell death in many cell types of different origins. The potential role of cytostatic agents in the apoptotic process of the inner ear needs, however, to be confirmed in large-scale specimens from patients treated with genotoxins. The fact, however, that apoptotic cells are also seen in association with ActD indicates that the fine sensory structure of the cochlea may also be a target for certain chemotherapeutic agents when administered in high doses.
Asunto(s)
Envejecimiento/patología , Apoptosis , Cóclea/citología , Adulto , Anciano , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Biomarcadores/análisis , Bleomicina/farmacología , Cisplatino/farmacología , Cóclea/química , Cóclea/embriología , Fragmentación del ADN , Células Epiteliales/química , Células Epiteliales/citología , Femenino , Regulación del Desarrollo de la Expresión Génica , Técnicas Genéticas , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Órgano Espiral/citología , Órgano Espiral/embriología , Adhesión en Parafina , Antígeno Nuclear de Célula en Proliferación/análisis , Transglutaminasas/análisis , Proteína p53 Supresora de Tumor/análisis , Vestíbulo del Laberinto/química , Vestíbulo del Laberinto/citología , Vestíbulo del Laberinto/embriología , Vinblastina/farmacologíaRESUMEN
We have previously demonstrated that local tumor irradiation effectively enhanced the therapeutic effect of interleukin 2 (IL-2) therapy in an experimental murine renal adenocarcinoma model. Based on these preclinical studies, we have designed and initiated a Phase II trial of irradiation combined with IL-2 for the treatment of metastatic renal cell carcinoma. Patients received 800 cGy to the primary or metastatic lesions on days 1 and 15 followed by IL-2 (600,000 IU/kg i.v.) every 8 h on days 4-8 and 18-22. Sixteen patients were entered; all completed treatment and are therefore evaluable for toxicity and response. Two partial remissions were seen for a response rate of 12.5% (95% confidence interval, 0-28.7). There was no increase in toxicity over that which is anticipated from IL-2 alone. The antitumor activity seen in this trial is consistent with what would be expected from high-dose IL-2 alone.
Asunto(s)
Antineoplásicos/uso terapéutico , Carcinoma de Células Renales/tratamiento farmacológico , Carcinoma de Células Renales/radioterapia , Interleucina-2/uso terapéutico , Neoplasias Renales/tratamiento farmacológico , Neoplasias Renales/radioterapia , Adulto , Anciano , Carcinoma de Células Renales/patología , Terapia Combinada , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Neoplasias Renales/patología , Masculino , Persona de Mediana Edad , Metástasis de la NeoplasiaRESUMEN
The signal transduction of the formyl-Met-Leu-Phe (FMLP) receptor in polymorphonuclear leukocytes (PMNLs) from patients with non-insulin-dependent diabetes mellitus (NIDDM) was compared to that of PMNLs obtained from healthy volunteers. According to our previous studies in this group of patients neither the decrease in insulin binding capacity nor the enhanced insulin-degrading enzyme activity was involved. In control PMNLs, 10 nM FMLP induced a pertussis toxin-sensitive increase in phosphatidyl inositol (PI) cleavage and a subsequent Ca2+ signaling from the intracellular pools. On the other hand, the FMLP-induced protein kinase C (PKC) activation and translocation into the membrane could not be detected in these cells via the measurement of 32P incorporation into histone. In contrast, in PMNLs of this special group of patients suffering from NIDDM the FMLP stimulus produced a significantly low increase in PI cleavage and Ca2+ signaling from the intracellular pools. Moreover, in resting PMNLs of these patients with NIDDM, not only the [Ca2+]i but also the membrane-bound PKC activity was found to be significantly increased. In addition, PKC translocation into the cell membrane of diabetic PMNLs could be further triggered with FMLP as judged by the measurement of 32P incorporation into histone. Based on these results, it appears that the signaling of FMLP receptors in PMNLs of some NIDDM patients may have an alternative pathway through Ca2+ influx from extracellular medium, arachidonic acid cascade, and PKC activation.
Asunto(s)
Membrana Celular/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , N-Formilmetionina Leucil-Fenilalanina/farmacología , Neutrófilos/metabolismo , Receptores Inmunológicos/metabolismo , Receptores de Péptidos/metabolismo , Transducción de Señal , Ácido Araquidónico/metabolismo , Calcio/fisiología , Células Cultivadas , Diabetes Mellitus Tipo 2/inmunología , Activación Enzimática/efectos de los fármacos , Humanos , Fosfatos de Inositol/metabolismo , Insulina/metabolismo , Masculino , Persona de Mediana Edad , Neutrófilos/efectos de los fármacos , Toxina del Pertussis , Proteína Quinasa C/metabolismo , Receptor de Insulina/metabolismo , Receptores de Formil Péptido , Receptores Inmunológicos/efectos de los fármacos , Receptores de Péptidos/efectos de los fármacos , Estallido Respiratorio/efectos de los fármacos , Superóxidos/metabolismo , Factores de Virulencia de Bordetella/farmacologíaRESUMEN
In situ distribution of CD2+ T-lymphocytes, CD4+ and CD8+ T-cell subsets, CD14+ macrophages, interleukin-2 receptor alpha-chain (IL-2R alpha) and class II major histocompatibility complex antigen (major histocompatibility complex class II, HLA-DR) expressing cells were determined in 14 chronic human periapical granulomas by immunohistochemical method using monoclonal antibodies. CD2+ lymphocytes were rather evenly distributed within the classical granulation tissue and comprised 55% of the mononuclear cells. Macrophages were distributed all over the periapical area, but their proportion was much less than that of T lymphocytes. Both small, lymphocyte-like mononuclear cells and larger mononuclear cells resembling macrophages displayed mild to strong circumferential staining with the anti-HLA-DR antibody. The majority of lymphocytes expressed IL-2R alpha indicating the activated state of T cells within the lesion.
Asunto(s)
Granuloma Periapical/inmunología , Receptores de Interleucina-2/análisis , Anticuerpos Monoclonales , Antígenos CD2/análisis , Relación CD4-CD8 , Enfermedad Crónica , Antígenos HLA-DR/análisis , Humanos , Inmunohistoquímica , Activación de Linfocitos , Macrófagos , Receptores de Antígenos de Linfocitos T/análisis , Subgrupos de Linfocitos TRESUMEN
We have demonstrated that tumor irradiation enhanced the therapeutic effect of interleukin 2 (IL-2) on pulmonary metastases from a murine renal adenocarcinoma, Renca. To investigate the mechanism of interaction between tumor irradiation and IL-2 therapy, we have histologically evaluated the effects of each therapy alone or in combination on Renca pulmonary metastases. Following treatment of established lung metastases with irradiation and IL-2 therapy, lung sections were processed for H&E or immunohistochemical staining. We found that tumor irradiation or IL-2 therapy locally induced vascular damage, resulting in multifocal hemorrhages and mononuclear cell mobilization in the lung tissue. This effect was amplified in lungs treated with the combined therapy. Immunohistochemistry showed that irradiation produced a macrophage influx into irradiated tumor nodules, and systemic IL-2 therapy induced T-cell infiltration in tumor nodules. Lungs treated with the combined therapy exhibited massive macrophage, T-cell, and natural killer cell mobilization in disintegrating tumor nodules and in the lung tissue. This combined therapy caused a decrease in the number of proliferating tumor cells and an increase in the number of apoptotic cells, which were more marked than with either therapy alone. We suggest that the macrophages mobilized by radiation-induced tissue injury could play a role in phagocytosis of apoptotic tumor cells, processing and presenting of tumor antigens for a systemic immune response activated by IL-2. Tumor destruction may result from the concomitant action of activated T cells, natural killer cells, and macrophages infiltrating the tumor nodules.
Asunto(s)
Carcinoma de Células Renales/terapia , Interleucina-2/uso terapéutico , Neoplasias Renales/terapia , Animales , Carcinoma de Células Renales/patología , Carcinoma de Células Renales/rehabilitación , División Celular/efectos de los fármacos , División Celular/efectos de la radiación , Terapia Combinada , Inmunohistoquímica , Neoplasias Renales/patología , Neoplasias Renales/rehabilitación , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/patología , Pulmón/citología , Pulmón/efectos de los fármacos , Pulmón/patología , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos BALB CRESUMEN
We have previously demonstrated that local tumor irradiation effectively enhanced the therapeutic effect of IL-2 therapy on pulmonary metastases from a murine renal adenocarcinoma, Renca. Irradiation with 300 rad to the left lung only, followed by systemic IL-2 therapy, results in increased tumor reduction in both lungs, suggesting that radiation enhances the systemic effect of immunotherapy. In this study, we show that irradiation of the tumor-bearing organ is essential for the combined effect of both modalities. This effect is radiation dose-dependent as increases in the radiation dosage result in greater tumor reduction in the irradiated field as well as systemically in nonirradiated fields when combined with immunotherapy. We find that irradiation has a direct inhibitory effect on Renca cell growth in vitro. Irradiation of Renca cells also causes an upregulation in H-2Kd class I MHC antigen detectable at 300 rad and more pronounced with 800 rad. By in vivo selective depletion of lymphocyte subsets, we demonstrate the involvement of Lyt-2+ and L3T4+ T cell subsets and AsGM1+ cells, including NK cells, in the antitumor effect mediated by tumor irradiation and IL-2 therapy. Immunohistochemistry studies, performed on lung sections, showed a significant infiltration of CD3+ T cells and macrophages in the tumor nodules following treatment with tumor irradiation and IL-2 therapy. Our studies indicate that the mechanism of interaction between tumor irradiation and immunotherapy may include radiation-induced alterations in the tumor growth and antigenicity which may enhance or trigger an anti-tumor response elicited by IL-2 and mediated by T cells, AsGM1+ cells, and macrophages.
Asunto(s)
Adenocarcinoma/terapia , Carcinoma de Células Renales/terapia , Interleucina-2/uso terapéutico , Neoplasias Renales/terapia , Pulmón/efectos de la radiación , Adenocarcinoma/inmunología , Animales , Carcinoma de Células Renales/inmunología , División Celular/efectos de la radiación , Terapia Combinada , Relación Dosis-Respuesta en la Radiación , Femenino , Antígenos H-2/análisis , Neoplasias Renales/inmunología , Células Asesinas Naturales/inmunología , Depleción Linfocítica , Ratones , Ratones Endogámicos BALB CRESUMEN
Alveolar macrophages protect the lungs against noxious agents. Proteases and peptidases are essential for this defense and many metabolic activities. Human alveolar macrophages were evaluated for the presence of six important peptidases. Deamidase, a serine peptidase identical with the lysosomal protective protein and possibly with cathepsin A, had high specific activity in alveolar macrophages and is also present in cultured mouse J774A.1 and human U937 cells, used for the sake of comparison. In fractionated J774A cells, most of the deamidase activity was in the lysosomal fraction and in the final supernatant. Deamidase in human alveolar macrophages, obtained by bronchoalveolar lavage from 23 patients, cleaved dansyl-Phe-Leu-Arg at a rate of 2.26 mumol/h/mg protein and hydrolyzed the chemotactic peptide N-f-Met-Leu-Phe even faster, at a rate of 53.1 mumol/h/mg protein, the highest activity for this enzyme with any of the cells we tested. Rabbit antiserum, elicited with the recombinant partial sequence of the enzyme, immunoprecipitated 77-88% of the macrophage deamidase. In immunocytochemistry, this antiserum localized deamidase within the human macrophages. The enzyme was inhibited by diisopropylfluorophosphate (DFP; 1 mM) and by ebelactone B (10 microM), noncompetitively. The mRNA of deamidase was detected in mouse macrophages by Northern blot; the two protein chains of deamidase were shown in human macrophages by Western blot. In addition, two other serine peptidases were also highly active in macrophages: dipeptidyl peptidase IV (1.38 mumol/h/mg protein) and prolylcarboxypeptidase (0.72 mumol/h/mg protein). The activity of plasma membrane zinc metallopeptidases, neutral endopeptidase 24.11 and carboxypeptidase M, in contrast, was low or absent (angiotensin I converting enzyme; kininase II).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Carboxipeptidasas/metabolismo , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/metabolismo , Lisosomas/enzimología , Macrófagos Alveolares/enzimología , Secuencia de Aminoácidos , Animales , Northern Blotting , Western Blotting , Carboxipeptidasas/antagonistas & inhibidores , Catepsina A , Células Cultivadas/enzimología , Factores Quimiotácticos/metabolismo , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/antagonistas & inhibidores , Humanos , Hidrólisis , Inmunohistoquímica , Proteínas de la Membrana/metabolismo , Ratones , Datos de Secuencia Molecular , Péptidos/metabolismo , Pruebas de Precipitina , Especificidad por SustratoRESUMEN
Local tumor irradiation enhances the effect of interleukin-2 (IL-2) therapy in the Renca murine renal adenocarcinoma model. To investigate the mechanism(s) of this interaction, we studied the in vitro and in vivo effects of irradiation on the tumor cells. Tumor cells from in situ irradiated renal tumors had diminished proliferation in vitro. A similar growth inhibition was noted following injection of irradiated Renca cells into naive mice, but this effect could be overcome by injecting more cells. Histologic evaluation of tumors derived from irradiated cells revealed a decrease in mitosis and an increase in multinucleated giant cells, apoptosis and micronecrosis. The presence of irradiated tumor reduced the growth of nonirradiated tumor cells when both were injected into separate flanks of the same animal, suggesting that irradiated tumor cells may trigger a systemic antitumor response. Interleukin-2 therapy given after injection of irradiated tumor cells caused a significant increase in leukocytic infiltrates and micronecrosis. Our findings indicate that radiation directly affects tumor growth and induces a systemic mechanism which could be enhanced by IL-2.
Asunto(s)
Carcinoma de Células Renales/patología , Interleucina-2/uso terapéutico , Neoplasias Renales/patología , Animales , Carcinoma de Células Renales/secundario , Carcinoma de Células Renales/terapia , División Celular/efectos de los fármacos , División Celular/efectos de la radiación , Femenino , Inmunoterapia , Interleucina-2/farmacología , Neoplasias Renales/terapia , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos BALB C , Radioterapia/métodosRESUMEN
This study was designed to test the specific binding to human ovarian serous adenocarcinomas of a drug-antibody conjugate [daunorubicin (DNR-OC-125], made from a new analog (PIPP-DNR) of daunorubicin that chemically links the drug to monoclonal antibodies. We recently reported that the DNR-OC-125 conjugate is selectively toxic in vitro to dividing cell populations of the human ovarian cancer cell lines SK-OV-3 and OVCAR-3 that express the CA-125 antigen [F. Sweet, L. O. Rosik, G. M. Sommers, and J. L. Collins, Gynecol. Oncol. 34, 305-311 (1989)]. In the present study, immunofluorescence data show that the DNR-OC-125 conjugate has high affinity and specificity for proliferating malignant cells from human ovarian tumors. The results demonstrate that the DNR-OC-125 conjugate retains the specific binding to CA-125 antigenic sites characteristic of the OC-125 monoclonal antibody moiety. The DNR-OC-125 conjugate selectively binds to CA-125 antigen-positive ovarian cancerous tissue in both cryostat and paraffin-embedded tissue sections. This is consistent with the earlier in vitro data from dividing populations of two human ovarian cancer cell lines that revealed retention by the DNR-OC-125 conjugate of both the specificity due to OC-125 and the cytotoxicity of daunorubicin. The present immunofluorescence studies in the DNR-OC-125 conjugate is tested on human ovarian serous tumors indicate that the OC-125 monoclonal antibody can indeed serve as a cancer-targeting carrier for daunorubicin and its analogs.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores/análisis , Cistadenocarcinoma/metabolismo , Daunorrubicina/metabolismo , Inmunotoxinas/metabolismo , Neoplasias Ováricas/metabolismo , Anticuerpos Monoclonales/metabolismo , Antígenos de Carbohidratos Asociados a Tumores/inmunología , Cistadenocarcinoma/inmunología , Portadores de Fármacos , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Neoplasias Ováricas/inmunologíaRESUMEN
Mesenchymal renal tumors in F-344 newborn rats were induced by a single dose of dimethylnitrosamine. The induced tumors were successfully transplanted into adult rats under the renal capsule. Neither the primary nor the transplanted neoplasms from various generations of grafts changed their morphological features during the tumor passage, having the same cellularity with high mitotic activity and the tendency to invade the host kidney rapidly. On the basis of lectin histochemistry and immunohistology, the tumor proved to be a mesenchymal neoplasm without any obvious capacity of the proliferating cells to differentiate into any well-known organoid element normally found in mature renal parenchyma. However, the proliferating neoplastic cells were found to have a strong vimentin positivity with desmin expression. Ultrastructurally, myofilaments with attachment bodies characteristic of smooth muscle cells were generally present in various amounts in many tumor cells. In addition, on the basis of the physiological data and on kidney/tumor renin activity obtained, it is interesting to note that the tumor-graft-invaded kidneys retained their enzyme activity, despite the obvious loss of renal tissue including glomeruli. However, the immunohistochemical findings with anti-renin antibody have clearly shown that this is not due to a renin-producing tumor but rather to the surviving (probably) non-neoplastic arterioles retaining the capacity to produce renin. Although these arterioles have mostly been found next to necrotic areas, commonly occurring in dimethylnitrosamine-induced transplantable renal tumors, the question of a possible physiological role of renin in tumor necrosis or in angiogenesis has remained open.
Asunto(s)
Dimetilnitrosamina , Neoplasias Renales/patología , Mesenquimoma/patología , Animales , Animales Recién Nacidos , Femenino , Neoplasias Renales/análisis , Neoplasias Renales/inducido químicamente , Masculino , Mesenquimoma/análisis , Mesenquimoma/inducido químicamente , Trasplante de Neoplasias , Ratas , Ratas Endogámicas F344 , Renina/análisis , Organismos Libres de Patógenos Específicos , Factores de TiempoRESUMEN
Authors survey the cyto-biological effect of extra-renal renin-angiotensin system on the basis of literature data and of their own previous results. It is established that renin and angiotensins in extra-renal localisation take part mainly in inflammatory process. In this respect, one of the important target cell group of angiotensin system is the certain elements of mononuclear phagocyte system, on which angiotensin II has cytokine-like effect. Renin-angiotensins detected by authors in non-activated alveolar mono-phages and monocytes raise the possibility, that these cells also have independent, intra-cellular regulating renin-angiotensin.
Asunto(s)
Fagocitos/fisiología , Sistema Renina-Angiotensina/fisiología , Factores Biológicos/fisiología , Citocinas , Humanos , Leucocitos Mononucleares/fisiologíaRESUMEN
The presence of angiotensins was demonstrated in normal unstimulated alveolar macrophages and monocytes from both mice and rats. These peptides were partially purified from cell homogenates by ion exchange chromatography and identified as being [Ile5] angiotensin I (Ang I), [Ile5] angiotensin II (Ang II) and to a lesser extent [Ile4] angiotensin III (Ang III) using high performance liquid chromatography (HPLC). Based on the present data both alveolar macrophages and monocytes expressed Ang I as quantified by a specific and sensitive radio-immunoassay (RIA) from the HPLC eluates. In contrast to this, alveolar macrophages from both mice and rats exhibited a fairly low, if detectable Ang II content. It seems reasonable to suggest that, in contrast to monocytes, macrophages do not generate and/or incorporate Ang II appreciably, at least in their resting stage. Although it is still not obvious whether these mononuclear phagocytes generate or simply capture angiotensin(s) from the blood pool or from the tissues; they must serve as in vivo target cells for the angiotensin system, at least for the plasma or tissue clearance of these molecules.