RESUMEN
Bullous pemphigoid (BP) is the most frequent autoimmune bullous skin disease, characterised by auto-antibodies against the hemidesmosome complex. Recently, regulatory T cells (Tregs) have been implicated in the development of several autoimmune diseases; few data are available in BP, failing to demonstrate a role of this subset in disease pathogenesis. The aim of this study was to investigate the expression and phenotypes of different Tregs (CD4+ CD25brightFOXP3+ and CD8+ CD28- cells) in BP to clarify whether the depletion of this subset constitutes one mechanism of tolerance loss. The CD4+ CD25brightFOXP3 and CD8+ CD28- circulating subsets were determined by flow-cytometry in 26 untreated BP patients and compared with a group of age- and sex-matched healthy controls (HC, n = 30). Absolute and percentage values of the CD4+ CD25brightFOXP3+ cells were significantly reduced in BP compared with HC (median CD25brightFOXP3+ expression within CD4+ cells: 1.8 vs. 3.5%, p = 0.002); conversely, BP patients were characterised by a significant expansion of the CD25brightFOXP3- "activated" T-cell subset. CCR4 and CD62L were expressed on the majority of CD4+ CD25brightFOXP3+ cells (75.2 and 82.3%, respectively). No differences in the CD8+ CD28- subset were found between BP and HC. This is the first report showing a significant reduction of circulating CD4+ CD25brightFOXP3+ Treg frequency in BP patients.
Asunto(s)
Penfigoide Ampolloso/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T Reguladores/inmunología , Anciano , Anciano de 80 o más Años , Circulación Sanguínea/inmunología , Antígenos CD4/metabolismo , Separación Celular , Femenino , Citometría de Flujo , Factores de Transcripción Forkhead/metabolismo , Humanos , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: Psoriasis is sustained by pro-inflammatory CD4+ T helper cells mainly belonging to the Th1, Th17 and Th22 lineage. OBJECTIVE: To identify whether treatment with the anti-tumour-necrosis-factor antagonist etanercept is able to induce significant modulations in transcription factor and cytokine mRNA gene expressions related to the different T cell immune response polarization (Th1, Th2, Th17 and regulatory T cells, Treg and to correlate them with clinical response. METHODS: The study population included 19 psoriasis patients treated with etanercept and 19 healthy subjects. Blood samples were collected at baseline and every 4 weeks during treatment. Taqman quantitative real-time polymerase chain reaction was applied to analyse the expression of: Stat-4, T-bet, IL-12p35 and IFN-γ (Th1-related); GATA-3, IL-4 (Th2-related); Stat-3, RORγt, IL-23p19 (Th17-related); Foxp3, IL-2 (Treg-related). Flow cytometry was applied to analyse CD4+CD25+(bright)Foxp3+ cells in peripheral blood. RESULTS: Upregulation of Th1 and Th17 and downregulation of Treg subsets was found at baseline. The response to etanercept could be associated with a significant reversal of the Th1/Th17 activation, and a concomitant upregulation of Th2 and Treg subsets. CONCLUSION: Our data may contribute to a better understanding of the mechanisms underlying the achievement of clinical response in psoriasis and could be helpful for the identification of early predictive markers of response.
Asunto(s)
Inmunoglobulina G/uso terapéutico , Factores Inmunológicos/uso terapéutico , Psoriasis/tratamiento farmacológico , Receptores del Factor de Necrosis Tumoral/uso terapéutico , Linfocitos T Reguladores/metabolismo , Células TH1/metabolismo , Células Th17/metabolismo , Células Th2/metabolismo , Adulto , Estudios de Casos y Controles , Citocinas/genética , Etanercept , Femenino , Perfilación de la Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Psoriasis/inmunología , ARN Mensajero/metabolismo , Factores de Transcripción/metabolismo , Resultado del TratamientoRESUMEN
BACKGROUND: Systemic sclerosis (SSc) and morphoea are connective tissue diseases characterized by fibrosis of the skin. Although to date their pathogenesis has not been clearly defined, it is thought that autoimmunity may play a role in the development of the skin lesions observed in both these diseases. As regulatory T cells (Tregs) play a key role in the modulation of immune responses, it has recently been suggested that Treg impairment may lead to the development of autoimmune diseases. OBJECTIVES: To investigate the presence of Tregs and their immunomodulatory cytokines, transforming growth factor (TGF)-beta and interleukin (IL)-10, in patients with SSc and morphoea. PATIENTS/METHODS: Fifteen patients with SSc and 15 with morphoea were enrolled. Immunohistochemistry was applied to identify FoxP3+ (forkhead/winged helix transcription factor) Tregs, TGF-beta+ cells and IL-10+ cells in the skin, cytofluorometry to detect CD4+CD25+FoxP3+ Tregs in the blood, and enzyme-linked immunosorbent assays to analyse TGF-beta and IL-10 serum levels. RESULTS: Fewer FoxP3+ Tregs and TGF-beta+ and IL-10+ cells were found in the skin of patients with scleroderma than in controls. Similarly, there were reduced TGF-beta and IL-10 serum levels and fewer circulating CD4+CD25brightFoxP3+ cells in patients with SSc or morphoea, than in controls. CONCLUSIONS: The quantitative reduction of Tregs, together with that of TGF-beta and IL-10 serum levels, may be responsible for the loss of tolerance observed in both SSc and morphoea.
Asunto(s)
Esclerodermia Sistémica/inmunología , Piel/inmunología , Linfocitos T Reguladores/inmunología , Adulto , Recuento de Linfocito CD4 , Femenino , Factores de Transcripción Forkhead/análisis , Humanos , Interleucina-10/análisis , Interleucina-10/sangre , Subunidad alfa del Receptor de Interleucina-2/análisis , Masculino , Persona de Mediana Edad , Esclerodermia Localizada/inmunología , Factor de Crecimiento Transformador beta/análisis , Factor de Crecimiento Transformador beta/sangreRESUMEN
BACKGROUND: Regulatory T-cell (T(reg)) modulation is one of the potential mechanisms of anti-tumour-necrosis-factor biological agents. However, literature data on psoriasis patients are lacking. OBJECTIVE: To analyse the circulating CD4+CD25(bright)FOXP3+ subset in 30 patients with psoriasis vulgaris/arthropathic psoriasis treated with biologicals and to investigate its relationship with the clinical response. METHODS: The CD25(bright)FOXP3+ expression within the CD4+ subset was determined by multi-parameter flow cytometry at baseline and during treatment. FOXP3 mRNA expression was analysed by real-time reverse transcription PCR. RESULTS: A response was obtained in 16/17 patients (91.1%) with increased CD25(bright)FOXP3+ values and in only 3/11 patients (27.3%) who showed a CD25(bright)FOXP3+ decrease during biological treatment (p = 0.0001). Responders showed significantly higher values than did non-responders as from the first 2 months of treatment (p = 0.0032). A significantly higher posttreatment expression of mRNA FOXP3 was observed in responders compared to non-responders. CONCLUSION: Biological drugs induce a circulating T(reg) up-regulation in a significant percentage of patients; such an increase is an early predictive marker of response.
Asunto(s)
Antígenos CD4/inmunología , Factores de Transcripción Forkhead/inmunología , Subunidad alfa del Receptor de Interleucina-2/inmunología , Psoriasis/inmunología , ARN Mensajero/genética , Linfocitos T/inmunología , Regulación hacia Arriba/genética , Adulto , Anciano , Factores Biológicos/uso terapéutico , Femenino , Citometría de Flujo , Factores de Transcripción Forkhead/biosíntesis , Factores de Transcripción Forkhead/genética , Humanos , Masculino , Persona de Mediana Edad , Psoriasis/tratamiento farmacológico , Psoriasis/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Resultado del TratamientoRESUMEN
The mechanisms of action of extracorporeal photochemotherapy (ECP) in cutaneous T-cell lymphoma (CTCL) are poorly understood. Recently, ECP has been shown to induce an increase in regulatory T cell (Treg) expression and functional activities in Graft-versus-host-disease (GvHD), whereas no data are available in CTCL patients. The aim of this study is to evaluate whether ECP is able to modulate the expression levels of the circulating CD4+CD25+bright subset in CTCL patients and whether these modifications are related to the disease course. The patient population included 43 CTCL and 15 chronic GvHD patients treated by ECP at our institutions since 1992. The expression of the circulating CD4+CD25+bright subset was analysed at baseline and sequentially during treatment by flow-cytometry. Fifty healthy donors were used as controls. The baseline circulating CD4+CD25+bright percentage values in CTCL (median: 4.3 percent) were similar to those of healthy donors, whereas GvHD showed significantly lower values (median: 1.5 percent; p<0.001). During treatment, CTCL patients were characterised by an early decrease (from 4.3 percent to 2.4 percent median after 6 months). The CD4+CD25+bright decrease was associated to the disease course, as it occurred in 91.3 percent of responding but in only 25 percent of PD patients (p=0.0001). On the other hand, a significant increase of CD4+CD25+bright cells was observed in GvHD. ECP induces a reciprocal modulation of the circulating CD4+CD25+bright cells in CTCL and GvHD, with a downregulation in CTCL potentially associated with the response mechanisms.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Enfermedad Injerto contra Huésped/terapia , Subunidad alfa del Receptor de Interleucina-2/análisis , Linfoma Cutáneo de Células T/terapia , Fotoféresis , Adulto , Anciano , Anciano de 80 o más Años , Recuento de Linfocito CD4 , Estudios de Casos y Controles , Enfermedad Crónica , Femenino , Citometría de Flujo , Enfermedad Injerto contra Huésped/inmunología , Humanos , Linfoma Cutáneo de Células T/inmunología , Masculino , Persona de Mediana Edad , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
BACKGROUND: Primary cutaneous T-cell lymphomas (CTCLs) are a heterogeneous group of lymphomas where the tumour population emerges within a multiple subclone pattern. Mycosis fungoides (MF) and Sézary syndrome (SS) are characterized by the expansion of clonal CD4+/CD45RO+ memory T cells. Lymphomatoid papulosis (LyP) is a chronic, lymphoproliferative disorder included in the CD30+ primary CTCL spectrum. Several studies have suggested a role of viral infection for super-antigenic activation of T lymphocytes; however, evidence of their association with CTCLs is still lacking. Human herpesvirus (HHV) 7 is a CD4+ T-lymphotropic herpesvirus; its restricted cellular tropism and the ability to induce cytokine production in infected cells could make it an important pathogenic cofactor in lymphoproliferative disorders. OBJECTIVES: To investigate the presence of HHV7 DNA on CTCL and healthy skin donors (HD). METHODS: We used quantitative real time polymerase chain reaction to evaluate the potential pathogenic role of HHV7. RESULTS: Twenty-seven of 84 (32.1%) HD were positive for HHV7 DNA. Twenty-one of 148 (14.2%) patients with CTCLs were positive for HHV7 DNA: nine of 39 (23.1%) SS, six of 14 (42.9%) CD30+ CTCLs and six of 24 (25.0%) LyP, and HHV7 DNA was negative in all 71 patients with MF. CONCLUSIONS: These results seem to exclude a pathogenic role of HHV7 in CTCLs, suggesting the possibility of skin as a latency site.
Asunto(s)
Herpesvirus Humano 7/aislamiento & purificación , Linfoma Cutáneo de Células T/virología , Neoplasias Cutáneas/virología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Genes Codificadores de la Cadena gamma de los Receptores de Linfocito T/genética , Herpesvirus Humano 7/genética , Humanos , Linfoma Cutáneo de Células T/patología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Neoplasias Cutáneas/patología , Linfocitos T/inmunología , Adulto JovenRESUMEN
BACKGROUND: Chronic dermatoses, as well as Sézary syndrome (SS), the erythrodermic and leukaemic cutaneous T-cell lymphoma, display a T-cell memory pattern. Recent findings suggest that different memory T-cell subsets can be recognized based on CD27 and CD45RO/RA expression. No data are reported as to CD27 expression in SS. OBJECTIVES: To evaluate different memory T-cell subsets, i.e. central memory (TCM), effector memory (TEM) and terminally differentiated cells in SS and inflammatory erythroderma (IE). MATERIALS AND METHODS: Forty SS and 137 IE patients were included. CD27 and CD45RO/CD45RA expression was analysed by flow cytometry on peripheral blood lymphocytes and immunohistochemistry. RESULTS: A significantly higher expression of the CD4+CD27+CD45RA- TCM subset was observed in SS whilst IE patients were characterized by increased CD4+CD27-CD45RA- TEM levels. The Vbeta-restricted population was homogeneously CD4+CD26-CD27+ in the SS subjects. CONCLUSIONS: SS and IE are characterized by a different memory T-cell subset expression; CD27 expression could be used as an additional diagnostic tool in the differential diagnosis.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Dermatitis Exfoliativa/inmunología , Síndrome de Sézary/inmunología , Subgrupos de Linfocitos T/inmunología , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/análisis , Anciano , Análisis de Varianza , Linfocitos T CD4-Positivos/metabolismo , Dermatitis Exfoliativa/diagnóstico , Dermatitis Exfoliativa/etiología , Diagnóstico Diferencial , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Memoria Inmunológica/inmunología , Inmunofenotipificación , Antígenos Comunes de Leucocito/análisis , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Valores de Referencia , Síndrome de Sézary/diagnóstico , Piel/inmunología , Piel/patología , Estadísticas no Paramétricas , Subgrupos de Linfocitos T/metabolismoRESUMEN
BACKGROUND: The finding of a suspicious urinary cytology is not uncommon in melanoma patients, in as much as morphology alone is often unable to distinguish the variable cytological features of melanoma cells. To date, although tyrosinase reverse transcription (RT)-PCR assay has been used to identify melanoma cells in peripheral blood and tissues, this method has not been applied to the analysis of urine samples. METHODS: RT-PCR mRNA tyrosinase expression was analysed in 79 urine samples from patients with metastatic melanoma and correlated with standard morphology/immunocytology. The results were compared with the disease course and presence of genito-urinary involvement. RESULTS: A positive RT-PCR expression was found in 18/79 urine samples from patients with metastases; four of the 18 patients had positive cytology, nine had atypical cytology, and five had negative cytology. Genito-urinary metastases were demonstrated in 27.8% tyrosinase-positive patients but in only 9.8% of the negative patients. The majority of tyrosinase-positive patients had a progressive disease unresponsive to chemotherapy. Urine samples from 20 patients with non-melanoma cancer and 20 healthy subjects were all negative. CONCLUSIONS: Our data demonstrate the higher sensitivity of RT-PCR compared with standard cytology in detection of urinary melanoma cells, and suggest that this assay could be used as an additional tool in the presence of negative or suspicious cytology.
Asunto(s)
Biomarcadores de Tumor/orina , Melanoma/diagnóstico , Melanoma/secundario , Neoplasias Urogenitales/diagnóstico , Neoplasias Urogenitales/secundario , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Citodiagnóstico/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Monofenol Monooxigenasa/sangre , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/orina , ARN Mensajero/genética , ARN Neoplásico/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad , Manejo de Especímenes/métodos , Urinálisis/métodosRESUMEN
BACKGROUND: A dominant T-cell clone can be detected by polymerase chain reaction (PCR) in 40-90% of cutaneous samples from patients with cutaneous T-cell lymphoma (CTCL). MATERIALS AND METHODS: From 1996 to 2003 we analysed 547 cutaneous biopsies performed to exclude CTCL (mycosis fungoides, MF/Sézary syndrome, SS). The final diagnosis was benign inflammatory disease (BID) in 353 samples (64.5%) and CTCL in 194 (35.5%). T-cell receptor (TCR)-gamma gene rearrangement was studied by using a multiplex PCR/heteroduplex (HD) analysis. The PCR results were correlated with the clinical picture, the histological pattern and the presence of T-cell lineage antigen loss, using univariate and multivariate logistic regression analyses. OBJECTIVE: To determine the sensitivity and specificity of the multiplex PCR/HD analysis and to identify which are the clinical, histopathological or immunophenotypical features significantly associated with a positive T-cell clonality. RESULTS: A clonality was demonstrated in 83.5% of CTCL and in 2.3% of BID (P < 0.001). A significantly higher percentage of clonal cases was associated with the cutaneous T-score (71.4% in T1, 76.1% in T2 and 100% in nodular and erythrodermic MF samples) and with the presence of a T-cell lineage antigen loss (93.9% vs. 77.4%). Moreover, clonality was closely related to an increase in the histopathological score (51.3% in the samples with a score < 5, compared with 92% in the lesions with > or = 5). No significant difference in the percentage of clonal cases was found between T1/T2 and T3/T4 lesions with a histopathological score > or = 5. The multivariate logistic regression showed that the density and extent of the cell infiltrate, the degree of epidermotropism and the presence of cytological atypia share an independent predictive value for clonality in T1/T2 samples, even if the highest odds ratios (3.6) were associated with the density of the cell infiltrate. The disease course of T1/T2 patients was analysed according to the PCR findings. All the PCR-negative patients showed a long-standing stable disease course; on the other hand, a disease progression occurred in 12/87 (13.8%) positive patients. CONCLUSIONS: The multiplex PCR/HD analysis is associated with a high diagnostic accuracy (92.7%) in CTCL patients. The finding of a clonal T-cell rearrangement is more closely associated with the histological pattern (in particular with the density and extent of the cell infiltrate) rather than with the MF cutaneous T-score or immunophenotype.
Asunto(s)
Reordenamiento Génico de la Cadena gamma de los Receptores de Antígenos de los Linfocitos T , Micosis Fungoide/genética , Síndrome de Sézary/genética , Neoplasias Cutáneas/genética , Linfocitos T CD8-positivos/inmunología , Distribución de Chi-Cuadrado , Células Clonales , Dermatitis/genética , Dermatitis/inmunología , Dermatitis/patología , Progresión de la Enfermedad , Análisis Heterodúplex , Humanos , Inmunofenotipificación , Antígenos Comunes de Leucocito/análisis , Modelos Logísticos , Micosis Fungoide/inmunología , Micosis Fungoide/patología , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Síndrome de Sézary/inmunología , Síndrome de Sézary/patología , Piel/inmunología , Piel/patología , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patologíaRESUMEN
Reverse transcription polymerase chain reaction (RT-PCR) of tyrosinase mRNA has been applied for the detection of melanoma cells in the peripheral blood, lymph nodes and bone marrow of melanoma patients. We evaluated the diagnostic accuracy of RT-PCR in comparison to standard cytology and immunocytochemistry (ICC) for the identification of melanoma cells in biological fluids other than blood. Tyrosinase expression was evaluated together with standard cytology and ICC (anti-S100, HMB-45 and Melan-A antibodies) in biological fluid samples collected from 17 melanoma patients according to the site of metastatic involvement or clinical suspicion (eight cerebrospinal fluid (CSF) samples; three pleural effusions; four ascites; one bile sample, one pericardial effusion); 17 samples collected from patients with non-melanoma metastatic cancer were used as controls. Tyrosinase expression in the biological fluid sample was compared with the expression determined at the same time in peripheral blood. Positive tyrosinase expression was found in 12/17 melanoma and 3/17 non-melanoma cancer patients. Cytology/ICC showed the presence of neoplastic cells in only 7/12 melanoma samples with positive tyrosinase expression: radiological evidence of disease involvement was found in all these patients (three meningeal, two pleural, two peritoneal). Clear-cut radiological evidence of disease involvement at the sampling site was found in the five patients with negative cytology/ICC and positive RT-PCR (one CSF; four serous membrane effusions); all patients died of disease progression within four months of sampling. The five patients who were negative for both cytology/ICC and RT-PCR did not show any clinical evidence of disease recurrence at the sampling site. Only five of the 12 metastatic patients with positive tyrosinase expression in biological fluid showed positivity for tyrosinase in the peripheral blood. These preliminary results suggest that the analysis of biological fluids other than blood could be considered as a new potential clinical field of application for the tyrosinase mRNA assay.
Asunto(s)
Líquidos Corporales/metabolismo , Pruebas Diagnósticas de Rutina/métodos , Melanoma/diagnóstico , Melanoma/enzimología , Monofenol Monooxigenasa/genética , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/genética , Femenino , Humanos , Masculino , Melanoma/sangre , Melanoma/genética , Persona de Mediana Edad , Monofenol Monooxigenasa/análisis , Monofenol Monooxigenasa/metabolismo , Metástasis de la Neoplasia/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Reverse transcription polymerase chain reaction (RT-PCR) of tyrosinase mRNA has been applied for the detection of melanoma cells in the peripheral blood, lymph nodes and bone marrow of melanoma patients. We evaluated the diagnostic accuracy of RT-PCR in comparison to standard cytology and immunocytochemistry (ICC) for the identification of melanoma cells in biological fluids other than blood. Tyrosinase expression was evaluated together with standard cytology and ICC (anti-S100, HMB-45 and Melan-A antibodies) in biological fluid samples collected from 17 melanoma patients according to the site of metastatic involvement or clinical suspicion (eight cerebrospinal fluid (CSF) samples; three pleural effusions; four ascites; one bile sample, one pericardial effusion); 17 samples collected from patients with non-melanoma metastatic cancer were used as controls. Tyrosinase expression in the biological fluid sample was compared with the expression determined at the same time in peripheral blood. Positive tyrosinase expression was found in 12/17 melanoma and 3/17 non-melanoma cancer patients. Cytology/ICC showed the presence of neoplastic cells in only 7/12 melanoma samples with positive tyrosinase expression: radiological evidence of disease involvement was found in all these patients (three meningeal, two pleural, two peritoneal). Clear-cut radiological evidence of disease involvement at the sampling site was found in the five patients with negative cytology/ICC and positive RT-PCR (one CSF; four serous membrane effusions); all patients died of disease progression within four months of sampling. The five patients who were negative for both cytology/ICC and RT-PCR did not show any clinical evidence of disease recurrence at the sampling site. Only five of the 12 metastatic patients with positive tyrosinase expression in biological fluid showed positivity for tyrosinase in the peripheral blood. These preliminary results suggest that the analysis of biological fluids other than blood could be considered as a new potential clinical field of application for the tyrosinase mRNA assay. (Int J Biol Markers 2005; 20: 11-7).
RESUMEN
Vascular endothelial growth factor (VEGF) is known to play a crucial role in the growth and metastatization of solid tumours. In cancer patients, high VEGF serum levels correlate with tumour status and prognosis, but to date few data have been reported concerning VEGF in melanoma patients. In the present study, immunoenzymatic and reverse transcription-polymerase chain reaction (RT-PCR) techniques were used to detect VEGF-165 serum levels and the presence of tyrosinase mRNA, respectively, in the peripheral blood of a cohort of 155 melanoma patients at different clinical stages (30 stage I, 40 stage II, 40 stage III and 45 stage IV; AJCC classification). Data were compared with both the extent of the disease and the clinical course. The aim was to assess the relationship between VEGF serum levels, the presence of detectable circulating melanoma cells and melanoma progression. A significant increase in VEGF serum levels was found in melanoma patients, in particular in those with metastatic disease; a higher incidence of relapses was found in stage I-III disease-free patients who showed an increase in VEGF during follow-up. VEGF serum levels were significantly higher in patients with detectable circulating melanoma cells than in those with negative tyrosinase mRNA expression. The finding of both an increase in VEGF and the presence of detectable melanoma cells during follow-up was associated with a relapse rate of 81%. The relapse rate was significantly lower when either of the two parameters were present separately. Multivariate analysis of both overall survival and time-to-progression selected baseline tyrosinase expression in peripheral blood but not VEGF serum levels as an independent prognostic factor.
Asunto(s)
Biomarcadores de Tumor/sangre , Factores de Crecimiento Endotelial/sangre , Péptidos y Proteínas de Señalización Intercelular/sangre , Linfocinas/sangre , Melanoma/sangre , Monofenol Monooxigenasa/sangre , Proteínas de Neoplasias/sangre , Adulto , Anciano , Estudios de Cohortes , Progresión de la Enfermedad , Supervivencia sin Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Tablas de Vida , Masculino , Melanoma/enzimología , Melanoma/mortalidad , Melanoma/patología , Persona de Mediana Edad , Análisis Multivariante , Metástasis de la Neoplasia , Estadificación de Neoplasias , Células Neoplásicas Circulantes , Pronóstico , ARN Mensajero/sangre , ARN Neoplásico/sangre , Recurrencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Supervivencia , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
Peripheral blood involvement in mycosis fungoides (MF) patients is more frequent in the advanced stages and is associated with a worse prognosis. We report a MF patient with limited patch lesions on her shoulders, upper chest and thighs (T2N0M0) and peripheral blood involvement. Clonality in the peripheral blood was demonstrated by the PCR assay and confirmed by the expansion of the same restricted variable region of the TCR beta-chain (vbeta17) expressed in the cutaneous infiltrate. The patient was treated with fludarabine achieving a complete hematological response followed by an early relapse, whilst the cutaneous lesions remained unchanged. The soluble interleukin-2 receptor levels showed a decrease from baseline levels down to normal values at hematological remission, followed by a further increase. The low sLex/CLA expression in the cutaneous lymphoid infiltrate could have given rise to a higher recruitment in the peripheral blood.
Asunto(s)
Micosis Fungoide/sangre , Micosis Fungoide/patología , Neoplasias Cutáneas/sangre , Neoplasias Cutáneas/patología , Vidarabina/análogos & derivados , Anciano , Femenino , Citometría de Flujo/métodos , Humanos , Inmunohistoquímica/métodos , Inmunosupresores/uso terapéutico , Leucocitos/citología , Linfocitos/patología , Microscopía Electrónica/métodos , Micosis Fungoide/inmunología , Fenotipo , Reacción en Cadena de la Polimerasa/métodos , Receptores de Interleucina-2/efectos de los fármacos , Hombro , Neoplasias Cutáneas/inmunología , Muslo , Tórax , Vidarabina/uso terapéuticoRESUMEN
Various enzymatic or mechanical methods have been proposed in the past to dissociate cells from different solid tissues. An automated mechanical disaggregation device (Medimachine) has recently been proposed. Unfortunately, most of these techniques are associated with a high cellular damage and a low cell recovery and are difficult to apply to skin biopsies. In this paper, we propose a combined enzymatic and mechanical method based on Medimachine, useful for the isolation of skin infiltrating T-lymphocytes from small cutaneous biopsies. As this method is easy and allows for a more correct qualitative and quantitative cytofluorimetric analysis of the lymphocyte subsets, it may be useful in the immunophenotyping of cutaneous T-cell lymphomas.