Priming effects of GM-CSF, IFN-gamma and TNF-alpha on human neutrophil inflammatory cytokine production.

Identifying and evaluating the priming agents for cytokine release by neutrophils might be helpful in controlling the innate immune response of the host. In the present study we examined the role of granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) as priming agents for interleukin (IL)-1beta, IL-6 and TNF-alpha production by stimulated neutrophils from control subjects and malignant melanoma patients. When the cells from controls and patients were preincubated with primer agents, opsonized zymosan-stimulated inflammatory cytokine production was enhanced. The major neutrophil-priming factor for IL-6 secretion by polymorphonuclear leukocytes (PMNs) in the control and patient groups was TNF-alpha. However, GM-CSF and IFN-gamma are also significant primers. GM-CSF priming was critical for the release of TNF-alpha from PMNs in control and melanoma patients. The ability of GM-CSF, IFN-gamma and TNF-alpha to serve as effective priming agents for inflammatory mediator production by PMNs revealed a new role for these cytokines in the innate immune response of the melanoma-bearing host.

TNF-alpha, IL-6 and their soluble receptor serum levels and secretion by neutrophils in cancer patients.

Simultaneous evaluation of cytokines and their soluble receptor production and the serum levels can be helpful in understanding the local and systemic immune response of a tumor-bearing host. In the present study we examined serum levels of TNF-alpha, IL-6 and their soluble receptors: sTNFRp55, sTNFRp75 and sIL-6R confronted with their production by the polymorphonuclear neutrophils (PMN) from cancer patients. Examinations were carried out in patients with adenocarcinoma breast cancer and squamous cell carcinoma of the oral cavity and related to the clinical course and to different phases of therapy. Secretion of IL-6, sTNFRp55 and sTNFRp75 by PMN appeared to be dependent on tumor type, clinical progression of disease as well as on therapy, suggesting a significant role of these cells at different phases of the immune response to cancer associated with these mediators. Changes in values of TNF-alpha, IL-6 and their soluble receptors in sera of both cancer groups, dependent on tumor type, clinical progression and cancer therapy, could have a diagnostic and prognostic role in cancer disease.

Effect of IL-15 on the secretion of IL-1beta, IL-1Ra and sIL-1RII by PMN from cancer patients.

In the present study, we demonstrate an effect of rhIL-15 on the simultaneous secretion of IL-1beta and its natural inhibitors IL-1Ra and sIL-1RII by human neutrophils isolated from normal and tumour-bearing hosts (oral cavity cancer and melanoma patients) compared with serum IL-15 levels. We found an rhIL-15 influence on IL-beta and IL-1Ra secreted by PMN from healthy controls. In contrast, the PMNs from cancer patients were not sensitive to rhIL-15 stimulation. However, we found a priming effect of rhIL-15 on IL-1beta production by LPS-stimulated cells in oral cavity cancer. We also found no effect on sIL-1RII release by PMN from cancer patients.

TNFRs and IL-6R transmembrane receptors expression and release of their soluble forms by neutrophils and mononuclear cells from cancer patients.

TNF-alpha and IL-6 are multipotential mediators involved in the control of many host's reactions to tumour. Their biological effects are mediated through the membrane-bound receptors (TNFRp55, TNFRp75 and IL-16R respectively) which can exist in soluble forms. In the present study we compared release of soluble sTNFRp55, sTNFRp75 and sIL-6R with expression of their membrane-bound on PMN and PBMC. Cells were isolated from patients with cancer diseases with a different location and histological classification. We have found that alterations of membrane-bound TNFRp75 expression and in the secretion of soluble TNFRp75 form, as opposed to other receptors examined, are characteristic features of neutrophils and mononuclear cells isolated from cancer patients. The similar changes observed in the expression of TNFRp75 by PMN and PBMC appear to confirm a significant role of PMN in the tumour response mediated by TNF-alpha. Furthermore, the altered membrane-bound TNFRp75 expression and sTNFRp75 secretion appear to depend on the tumour type.

Tumor necrosis factor-alpha and soluble tumor necrosis factor receptors in the culture supernatants of polymorphonuclear cells and peripheral blood mononuclear cells from cancer patients.

Recent clinical and experimental studies have focused on the measurement of cytokines and their regulators, produced by immunocompetent cells. Their estimation may be used as parameters for the immune potential of cancer patients. In the present study we studied the ability of unstimulated and lipopolysaccharide (LPS)-stimulated polymorphonuclear cells (PMN) and peripheral blood mononuclear cells (PBMC) from oral cavity cancer and breast cancer patients to release tumor necrosis factor alpha (TNF-alpha) and soluble tumor necrosis factor receptors (sTNFR). There were significant differences concerning the parameters examined for PMN and PBMC from cancer patients as compared with normal subjects. We found significantly higher concentrations of sTNF-R p75 than sTNF-R p55 in the cell-culture supernatants. The culture supernatants of cells from oral cavity cancer patients contained higher concentrations of TNF-alpha and lower concentrations of sTNF-R p55 and sTNF-R p75 in comparison with breast cancer cell supernatants. In contrast, cells from breast cancer patients secreted lower concentrations of TNF-alpha and higher concentrations of sTNF-R p55 and sTNF-R p75. Although PBMC secreted higher concentrations of mediators than PMN, the quantitative dominance of PMN in the peripheral blood suggests an essential role of these cells in the defense reactions controlled by TNF-alpha.

Soluble TNF receptors release by polymorphonuclear cells and the serum levels in breast cancer patients before and after treatment.

Soluble cytokine receptors by binding to circulating cytokines play an important role in the regulation of immune response of the host. Measurement of their release may be helpful in the evaluation of cytokine-mediated reactions in patients with cancer disease. Soluble tumor necrosis factor receptors (sTNF-Rs) in the culture supernatants of PMNs and in the sera obtained from patients with breast cancer were determined. The examinations were carried out before and after treatment involving surgery and chemotherapy. The highest concentrations of sTNFRp55 and sTNFRp75 were observed in the culture supernatants of PMNs from patients before any treatment, the lower concentrations of them were found in the supernatants of PMNs after treatment. Sera from breast cancer patients presented increased levels of sTNFRp55 and sTNFRp75. Surgery treatment resulted in higher concentrations of sTNFRp55 and sTNFRp75. Decreased sTNF-Rs serum levels were obtained in patients after adjuvant chemotherapy. Different ability of PMNs to the release of TNF-regulatory proteins-sTNFRs appears to confirm PMNs participation in TNF-mediated reactions of the host during malignant process via sTNF-Rs release.

[Activity of cathepsin D in some neoplasms of the gastrointestinal tract]. / Aktywnosc katepsyny D w niektórych nowotworach przewodu pokarmowego.

In the present paper we analyzed cathepsin D activity in digestive tract cancers. Cathepsin D activity was estimated in 10% homogenates of oesophageal cancer, gastric cancer and colon cancer tissues and in the blood serum and expressed as the amount of liberated tyrosine which was assayed acc. to Folin-Ciocalteau. Mean cathepsin D activities in neoplastic tissues and normal counterparts were as follows: oesophaged cancer (218.5 mM Tyr/1/2 h vs 145.0 mM Tyr/1/2 h), gastric cancer (285.4 mM Tyr/1/2 h vs 142.3 mM Tyr/1/2 h) and colon cancer (233.7 mM Tyr/1/2 h vs 159.5 mM Tyr/1/2 h). In all examined neoplastic tissues cathepsin D activity was almost too-fold higher than in the normal counterparts. Cathepsin D activity in the sera of cancer patients was too a lesser degree higher than in the sera of normal subjects. The data indicate that estimating of cathepsin D activity in the neoplastic tissues homogenates and in the blood serum may be of diagnostic value and may constitute an information which is complementary to the analysis of other tumor markers and histopathologic examination.

Evaluation of CA 15-3 serum levels in breast cancer patients.

The usefulness of post operative serum CA 15-3 determination was evaluated in a group of 57 breast cancer patients. CA 15-3 was assayed at baseline and every 3 months for a median follow-up of 34 months. Recurrence-free survival probability for patients with elevated serum CA 15-3 levels before surgery was 53.5% versus 88% for patients with normal serum CA 15-3 levels (p < 0.201). In 12 of the 15 patients with increased trend for CA 15-3 during follow-up, elevated CA 15-3 levels preceded (10 patients) coincided (2 patients) with the clinical detection of tumour recurrence, providing a predictive value of an increased trend of 80%. In the group of 10 patients who showed elevated serum CA 15-3 levels prior to the detection of recurrence, the median load-time was 5 months/(range 3-11 months). We concluded that CA 15-3 had prognostic value in breast cancer. Serial determinations of CA 15-3 was consistent with the treatment results.