Blood Cell Cultures Of Patients Research Articles

Оценка показателей секреции цитокинов и пролиферативной активности в культурах мононуклеарных клеток крови больных клещевыми инфекциями

Associations between the immune response and the severity of tick-borne infections are poorly understood. Objective. Assessment of the parameters of cytokines secretion, proliferative activity and cytogenetic instability in the mononuclear blood cell cultures of patients with tick-borne encephalitis febrile form and ixodid tick-borne borreliosis nonerythemal form at the onset of the disease. Patients and methods. We assessed 24 parameters in the phytohemagglutinin-stimulated mononuclear blood cell cultures from 25 patients with tick-borne encephalitis, 24 patients with ixodid tick-borne borreliosis, and from 14 healthy donors. Statistical analysis included the Mann–Whitney test, Pearson’s goodness-of-fit test, Spearman’s rank correlation, and ROC analysis. Results. The patients compared with the controls showed a suppression of the stimulated blast transformation, increases in the levels of the micronucleated T-lymphocytes and the secretion of not only pro-inflammatory cytokines – tumor necrosis factor (TNF)-α and interleukin (IL)-6, but IL-10 and IL-4 as well. Pyrexia levels were negatively correlated with the blast transformation and positively correlated with the micronucleated T-cells levels and pro-inflammatory cytokines secretion. The predictors for the diagnosis of tick-borne encephalitis compared with borreliosis were decreases in the blank and/or stimulated levels of blast transformation and the production of TNF-α, IL-6, interferon-γ as well as increases in the micronucleated T-cell levels and the secretion of IL-8 and IL-10. Conclusion. The results represent the features of the immune response reactions and its association with the severity of the clinical picture. Key words: blast transformation, ixodid tick-borne borreliosis, tick-borne encephalitis, micronucleus assay, mononuclear blood cells, phytohemagglutinin, cytokines

33537 Inhibitory effects of ME3183, a novel phosphodiesterase-4 inhibitor, on stimulants-induced production of proinflammatory cytokines and chemokines in whole blood cell cultures of patients with plaque psoriasis

Inhibition of phosphodiesterase-4 (PDE4) has been reported as a therapeutic target for inflammatory diseases. ME3183 is an orally available and selective inhibitor of PDE4, which showed various anti-inflammatory effects in nonclinical studies. We examined the inhibitory effects of ME3183 on stimulants-induced production of proinflammatory cytokines and chemokines in whole blood cell cultures of healthy volunteers and patients with plaque psoriasis (Ps). Tumor necrosis factor alpha (TNF-α), interleukin 17A, interferon-γ, monocyte chemotactic protein-1 and interferon gamma-induced protein-10 were measured in each subject’s whole blood cell cultures stimulated with lipopolysaccharide (LPS) or phytohemagglutinin (PHA) in the presence of ME3183 or apremilast. Correlation between the inhibition of TNF-α production and each clinical parameter of patients were statistically analyzed. ME3183 showed approximately 5- to 40-folds more potent inhibitory activities against production of cytokines than apremilast both in the whole blood cell cultures of Ps patients and healthy subjects. Interestingly, our correlation analysis suggested that high body surface area (%) and long-duration of psoriasis may be associated with the potency (IC50) of ME3183 against TNF-α production. ME3183 showed higher potency than apremilast in the inhibition of proinflammatory cytokines and chemokines production. ME3183 can be a new generation of PDE4 inhibitor with improved therapeutic effect, and the efficacy of ME3183 will be investigated in patients with Ps in the P2a study.

AN ESTIMATION OF FREQUENCY AND SPECTRUM OF CHROMOSOMAL ABERRATIONS IN PATIENTS WITH CONNECTIVE TISSUE DYSPLASIA IN CONDITIONS OF BLOOD CELLS CONTACT WITH NANOSTRUCTURED MATERIALS

In order to evaluate cytogenetic effect of nanocomposite materials used in traumatology and orthopedics an investigation of chromosomal aberration ins in the culture of lymphocytes short-term contacted with artificial material (nanostructured titanium samples of implants bearing bilateral calcium phosphate coating) in patients with dysplasia of connective tissue (DCT) and osteogenesis imperfecta (OI) has been conducted. A reduction of percent of aberrant cells having single or/and paired chromosomal breaks as compared with spontaneous mutagenesis (without model samples) was revealed under introduced biocompatible composites into blood cell cultures of patients with DCT or OI. Analysis of frequency and spectrum of chromosomal aberrations in blood cells can be considered as potential prognostic criterion and a predictor of implant’s efficacy in the surgical treatment of DCT and OI complications.

Effect of Surgical Treatment on Lymphoproliferation in Advanced Supraglottic Laryngeal Cancer

The capacity of cell immunity to act against tumor cells has been presented as a decisive influence in the prognosis of patients with cancer. The aim of this study was to evaluate lymphoproliferation in nonadherent peripheral blood cell cultures of patients with advanced supraglottic laryngeal cancer. Fourteen patients with advanced supraglottic laryngeal cancer were studied prospectively. Lymphoproliferation was quantified by adding 3H-thymidine and measured in counts/minute using liquid scintillation spectrometry. Based on the ratio between stimulated and baseline cultures, the proliferation index was calculated before and 236 +/- 18 days after the surgery. Lymphoproliferation was lower in patients than in healthy controls (P = .01) in the preoperative as well as in the late postoperative period (P = .006 and P = .02, respectively). However, there was no change from preoperative to late postoperative. Pre- and postoperative results show that patients with advanced supraglottic laryngeal cancer present lymphoproliferation diminished before the surgery, and in the late postoperative period, there was no recovery of immune capacity evaluated by lymphoproliferation measurement.

Selective suppression of cytokine secretion in whole blood cell cultures of patients with colorectal cancer.

We have investigated the secretion of interferon alpha (IFN-alpha), IFN-gamma, interleukin-1alpha (IL-1alpha), IL-1beta, IL-2 and tumour necrosis factor alpha (TNF-alpha) in whole blood cell cultures (WBCCs) of colorectal cancer patients upon mitogen stimulation. Whereas the values for IL-1beta and TNF-alpha remained virtually unchanged in comparison with healthy control subjects, WBCCs of colorectal cancer patients secreted significantly lower amounts of IFN-alpha (P < 0.005), IFN-gamma (P < 0.0001), IL-1alpha (P < 0.0001) and IL-2 (P < 0.05). This reduction correlated with the progression of the disease. The total leucocyte and monocyte population were almost identical in both groups. In contrast, a dramatic depletion of lymphocytes was observed in colorectal cancer patients, which affected both lymphocyte counts (P < 0.0005) and their distribution (P < 0.0001). Our results suggest a selective suppression of cytokines in colorectal cancer patients that is related to tumour burden. Several mechanisms might account for this phenomenon, one of which might be lymphocyte depletion.

Cytokine production (IL-6 and TNF alpha) in whole blood cell cultures of patients with systemic lupus erythematosus.

Whole blood cell culture has great advantage over isolated peripheral blood mononuclear cell culture, because it needs only small amounts of blood and is fast to perform. The current report focuses on the measurement of IL-6 and TNF alpha produced by peripheral blood monocytes of patients with systemic lupus erythematosus (SLE) in the whole blood cell culture system. After an initial triggering with lipopolysaccharide (LPS), a specific stimulus for monocytes, a decreased production of IL-6 relative to the controls was observed. Dividing our SLE patients according to treatment with corticosteroids, overall the IL-6 production was decreased in the patients treated with corticosteroids. TNF alpha production was comparable with normals, with the exception of an increased spontaneous production and using LPS stimulus of 4 pg/ml. In the patients treated with corticosteroids a decreased TNF production was observed, in contrast to the non-treated patients in which an increased TNF production was found compared with the controls using LPS doses higher than 62 pg/ml. The impaired acute phase reaction (APR) that has been described in the literature, might be explained by our observation of a decreased production of mainly IL-6. However, also this study showed that treatment has a strong impact on ex vivo IL-6 and TNF production.

Cytokine production in human whole blood cell cultures of patients with multiple sclerosis: Evidence for Th1 or Th2?

Cytokine production in human whole blood cell cultures of patients with multiple sclerosis: Evidence for Th1 or Th2?

Cytokine production in whole blood cell cultures of patients with crohn's disease and ulcerative colitis

Levels of the cytokines interleukin-1-alpha, -1-beta, and -2 (IL-1-alpha, IL-1-beta, IL-2), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) were measured in the mitogen-stimulated whole blood cell cultures from 96 patients with Crohn's disease (48 untreated, 12 treated with sulfasalazine, 36 treated with corticosteroids), 74 patients with ulcerative colitis (21 untreated, 25 treated with sulfasalazine, 28 steroid treated), and 360 healthy controls. The cytokines were measured 4 days after induction by a sensitive immunoenzyme assay. In the blood cell cultures of the untreated and sulfasalazine treated patients with Crohn's disease and ulcerative colitis higher levels of TNF-alpha, IL-1-alpha and IL-1-beta were found whereas IL-2 production was decreased and IFN-gamma-production was not significantly different as compared to the controls. Leukocytes of the corticosteroid-treated patients with both diagnoses showed a lower production of all measured cytokines compared to the untreated patients. The same results were obtained, when the somewhat different counts of mononuclear cells in the peripheral blood of the patients and controls were taken into account. The elevated production of proinflammatory cytokines in the blood cell cultures suggests a systemic immune activation in patients with inflammatory bowel disease.

Impaired cytokine production in whole blood cell cultures of patients with gynaecological carcinomas in different clinical stages

The production of the cytokines IFN-gamma, IL-1-alpha, IL-2 and TNF-alpha was investigated in mitogen-stimulated, whole blood cell culture from 239 untreated patients with primary gynaecological carcinomas (breast, cervix, ovary, endometrium), and 191 healthy female controls. The cytokines were measured in the 4-day post-induction supernatants by a sensitive enzymoimmunological assay. In the blood cell cultures of all four groups of cancer patients, significantly lower values of IFN-gamma (P < or = 0.001), IL-2 (P < or = 0.01) and IL-1 alpha (P < or = 0.01) were found as compared to the controls, although lymphocyte and monocyte counts were almost identical. Grouping the tumour patients into different clinical stages we could show in the four groups of carcinomas a gradual depression of the cytokine production according to growing tumour burden.

Impaired cytokine production in whole blood cell cultures of patients with urological carcinomas.

The production of the cytokines interferon gamma (IFN gamma), interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor alpha (TNF alpha) was investigated in the mitogen-stimulated whole blood cell culture media from 51 patients with urinary bladder carcinomas, 52 patients with renal carcinomas, 31 patients with prostatic carcinomas and 360 healthy controls. The cytokines were measured 4 days after induction by a sensitive enzymo-immunological assay. In the blood cell culture supernatants of the patients with urinary bladder carcinomas significant lower levels of IFN gamma (P < or = 0.001), IL-2 (P < or = 0.01) and TNF alpha (P < or = 0.05) were found as compared to the controls. Blood cells of patients with renal carcinomas had lower production of IFN gamma (P < or = 0.01), IL-2 (P < or = 0.001) and IL-1 alpha (P < or = 0.01), whereas the values of the total group of patients with prostatic carcinomas were not significantly different from those of the controls. Lymphocyte and monocyte counts were almost identical in the control and all tumor patient groups. When the patients with renal carcinomas and prostatic carcinomas were analyzed according to their different clinical stages we could show a gradual depression of the IFN-gamma levels, which was related to tumor burden.