RESUMEN
Progression of B-cell chronic lymphocytic leukemia (B-CLL) is linked to an abnormal immune system in the host. Recent studies have suggested that polymorphonuclear neutrophils (PMN) play a role in the malignancy process through release of a wide range of mediators, involving nitric oxide (NO). The aim of this study was to examine NO production by PMN and, for comparison of peripheral blood mononuclear cell (PBMC) confronted with the expression and concentration of inducible NO synthase (iNOS) in these cells derived from patients with B-CLL. Results obtained were analyzed according to Rai' staging systems. Our results have shown impaired production of NO by human neutrophils and mononuclear cells. Furthermore, higher expression of iNOS detected by western blot as well as increased concentrations iNOS estimated by ELISA in these cells were observed. We also found higher expression and concentration of iNOS in PMN and PBMC patients in III stage in comparison with patients in I stage of the disease. Additionally we demonstrated a lower production of superoxide anion by neutrophils of patients with B-CLL. Results obtained suggest that impaired NO production, despite of enhanced expression of iNOS, may have a favorable effect on anti-tumor response in patients with B-cell chronic lymphocytic leukemia.
Asunto(s)
Leucemia Linfocítica Crónica de Células B/enzimología , Leucocitos Mononucleares/metabolismo , Óxido Nítrico Sintasa de Tipo II/sangre , Óxido Nítrico/sangre , Adulto , Anciano , Técnicas de Cultivo de Célula , Regulación hacia Abajo , Regulación Leucémica de la Expresión Génica , Humanos , Leucemia Linfocítica Crónica de Células B/inmunología , Leucocitos Mononucleares/inmunología , Persona de Mediana Edad , Neutrófilos/inmunología , Neutrófilos/metabolismo , Óxido Nítrico Sintasa de Tipo II/química , SuperóxidosRESUMEN
In this study we estimated the expression of TLR2 and apoptosis of PMN in patients with Lyme disease. The cells were isolated from heparinized whole blood by Gradisol G gradient and incubated 18 h with rhIL-15 and LPS. Expression of TLR2 was estimated in lysates of PMN by western blot, apoptosis of PMN by immunofluorescent analysis. The results obtained revealed the higher expression of TLR2 in PMN and higher percentage of apoptosis PMN in patients with Lyme disease compared with control. We observed an effect of rhIL-15 on the increased expression of TLR2 in PMN and the increased survival of PMN isolated from patients with Lyme disease. These findings suggest that IL-15 has the ability to modulate of neutrophil response against Borrelia burgdorferi.
Asunto(s)
Apoptosis , Interleucina-15/metabolismo , Enfermedad de Lyme/sangre , Glicoproteínas de Membrana/metabolismo , Neutrófilos/metabolismo , Receptores de Superficie Celular/metabolismo , Adulto , Western Blotting , Estudios de Casos y Controles , Femenino , Técnica del Anticuerpo Fluorescente , Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Proteínas Recombinantes/metabolismo , Receptor Toll-Like 2 , Receptores Toll-LikeRESUMEN
Tumor necrosis factor (TNF) superfamily, involving membrane receptors and ligands are important for the growth and survival of leukemic B cells. In the present study levels of TNF-alpha, sTNFRp55, sTNFRp75 and sCD40 and sCD40L in the serum of patients with B-CLL before and after treatment were measured. In sera of patients before treatment increased concentrations of sCD40 and decreased concentrations of sCD40L were shown. Increased concentrations of TNF-alpha and sTNFRp75 and lack of changes in sTNFRp55 concentrations were also found. Results obtained suggest that the relationships between examined soluble form of TNF family proteins may influence the development of B-cell chronic lymphocytic leukemia. The used therapy with 2CdA and CMC led to a favorable effect for host through changes in the relations between sCD40 and sCD40L. It was also found that sCD40 and sCD40L serum concentrations, which are dependent on the clinical stage and used therapy, are more sensitive tumor markers than TNF-alpha and its soluble receptor in patients with B-CLL treated with 2CdA and CMC.
Asunto(s)
Leucemia Linfocítica Crónica de Células B/sangre , Factores de Necrosis Tumoral/sangre , Antígenos CD40/sangre , Ligando de CD40/sangre , Femenino , Humanos , Leucemia Linfocítica Crónica de Células B/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Receptores del Factor de Necrosis Tumoral/sangre , Factor de Necrosis Tumoral alfaRESUMEN
INTRODUCTION: The tumor-polymorphonuclear neutrophil (PMN) relationship can be altered by the release of toxic molecules, such as nitric oxide (NO). The aim of the present study was to examine the expression of the inducible synthase of NO (iNOS)and NO production by human neutrophils of patients with oral cavity cancer. For comparison we performed similar examinations in autologous peripheral blood mononuclear cells (PBMCs). MATERIAL/METHODS: PMNs and PBMCs were isolated from the whole blood of 27 patients with squamous cell carcinoma of the oral cavity. iNOS protein expression in these cells was detected by Western blot. Total nitrite as an indicator of NO concentrations in the culture supernatants and the serum of patients was measured using a colorimetric assay. RESULTS: The PMNs of oral cavity cancer patients showed a significantly lower intensity of iNOS expression than those of healthy controls. The PBMCs of patients showed a more intensive expression of iNOS than the PMNs, but a lower intensity than the PBMCs of the controls. The expression of iNOS in rhIL-6 and rhIL-15-stimulated PMNs and PBMCs of patients increased in comparison with unstimulated cells. We observed lower productions of NO by PMNs and PBMCs of patients than those of the control group. CONCLUSIONS: The results revealed that altered iNOS expression and NO production are more characteristic of PMNs than of PBMCs of patients with oral cavity cancer. Additionally, this study provided new information about IL-6 and IL-15 activity in a tumor-bearing host.
Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Neoplasias de la Boca/metabolismo , Neutrófilos/metabolismo , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/biosíntesis , Adulto , Carcinoma de Células Escamosas/inmunología , Estudios de Casos y Controles , Humanos , Técnicas In Vitro , Interleucina-15/farmacología , Interleucina-6/farmacología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Persona de Mediana Edad , Neoplasias de la Boca/inmunología , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Nitratos/sangre , Óxido Nítrico Sintasa de Tipo II , Proteínas Recombinantes/farmacologíaRESUMEN
It is known that the relationship between pro-angiogenic and anti-angiogenic factors is responsible for the presence and intensity of neoangiogenesis. The angiogenic factors are produced by tumour cells and/or by tumour-infiltrating inflammatory cells such as macrophages or polymorphonuclear leukocytes (PMN). In the present study we compared VEGF secretion with IL-18 and NO release by PMN derived from oral cavity cancer patients. Knowledge of the relationship between mediators above could help in better understanding the role of PMN in angiogenesis in this patient group. The results from culture supernatants of PMN were confronted with the serum levels of parameters examined. We found an interesting relationship between VEGF and IL-18 concentrations in the culture supernatants of PMN derived from patients with oral cavity cancer. High production of VEGF was associated with low production of IL-18 by PMN derived from patients before treatment. During examinations after treatment we found lower concentrations of VEGF and higher concentrations of IL-18 than those in the study before treatment. In contrast to VEGF and IL-18, the NO production by PMN of cancer patients, before and after treatment, was unchanged. We also demonstrated markedly elevated serum levels of VEGF as well as IL-18 according to the progression of the disease. Results obtained indicate that relations between VEGF and IL-18 released by PMN may promote neoangiogenesis and may be important for benign tumour cells to acquire metastatic phenotype in the early stage of oral cavity cancer. Furthermore, our results suggest that the concentrations of VEGF and IL-18 in the serum are sensitive tumour markers in this patient group before and after treatment.
Asunto(s)
Interleucina-18/sangre , Neoplasias de la Boca/sangre , Neutrófilos/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico/sangre , Factor A de Crecimiento Endotelial Vascular/sangre , Adulto , Células Cultivadas , Humanos , Persona de Mediana EdadRESUMEN
The aim of this study was to estimate the release of IL-6 by human neutrophils (PMN) and peripheral blood mononuclear cells (PBMC) in patients with Lyme disease confronted with the serum levels. The cells were isolated from whole blood of 25 patients and of 10 healthy donors and cultured in the presence of LPS. In the culture supernatants and serum the concentration of IL-6 with ELISA (BioSource) were measured. In patients we observed higher values of IL-6 released by unstimulated PMN and PBMC in compared with control. In contrast to control, we didn't observe increased the release of IL-6 by LPS-stimulated PMN and PBMC as compared to unstimulated cells. In the serum of patient we found increased the concentration of IL-6. The higher ability of PMN and PBMC from patients with Lyme disease to release of IL-6 and the lack response to additional stimulation indicate the activation of PMN and PBMC in vivo.