The effect of perioperative analgesic drugs omnopon and dexketoprofen on the functional activity of immune cells in murine model of tumor surgery.

We aimed to investigate the effect of perioperative analgesia with nonselective cyclooxygenase-2 inhibitor dexketoprofen and opioid drug omnopon on the functional activity of immune cells in tumor excision murine model. Lewis lung carcinoma cells were transplanted into hind paw of C57/black mice. On the 23th day tumor was removed. Analgesic drugs were injected 30 min before and once a day for 3 days after the surgery. Biological material was obtained a day before, 1 day and 3 days after the tumor removal. IFN-γ, IL-4, IL-10 and TGF-ß mRNA levels in splenic cells were assessed by quantitative real-time RT-PCR. Cytotoxic activity of splenocytes was estimated by flow cytometry. We found that in splenocytes of mice received opioid analgesia IL-10 mRNA level was increased 2.3 times on day one after the surgery compared to preoperative level (P < 0.05), while in dexketoprofen group this parameter did not change. IFN-γ gene expression level on day 3 after tumor removal was 40% higher in splenocytes of dexketoprofen treated mice as compared with omnopon treated animals (P < 0.05). Cytotoxic activity of splenocytes on day 3 postsurgery was (62.2 ± 2.4)% in dexketoprofen against (50.2 ± 3.3)% in omnopon group. In conclusion, perioperative analgesia with cyclooxygenase inhibitor dexketoprofen in contrast to opioid analgesia with omnopon preserves higher functional activity of murine immune cells in the experimental model of tumor surgery.

Antineoplastic drug NSC631570 modulates functions of hypoxic macrophages.

Hypoxia is an important factor in the macrophages microenvironment. Many physiological and pathological processes including solid tumor development are characterized by both low oxygen content and presence of macrophages. Tumor-associated hypoxia causes alternative polarization of macrophages in tumor tissue and transformation of these cells into the allies of a malignant neoplasm. The aim of the work was to investigate the effect of NSC631570, a cancer-selective drug that is known to selectively accumulate in the tumor tissue, on hypoxic macrophage function. Murine peritoneal macrophages (PMs) were subjected to hypoxia (3% O2). Nitrite level was assayed by the Griess reaction. Arginase activity was measured by colorimetric method. ROS generation and phagocytosis was estimated by flow cytometry. O2(-) generation was assayed by the NBT reduction method. HMGB1 expression was determined by ELISA. 42 h hypoxia caused alternative polarization of murine PMs with significant arginase prevalence. NSC631570 repolarized arginine metabolism of hypoxic macrophages to NOS dominant and activated their pro-inflammatory functions: recovered ROS production and increased alarmin release NSC631570 can restore pro-inflammatory functions of macrophages, alternatively polarized by hypoxia.

The effect of NSC-631570 (ukrain) alone and in combination with pathogen-associated molecules on cell cycle distribution and apoptosis induction of mouse melanoma cells with different biological properties.

UNLABELLED: Monotherapy and combined application of antitumor drug NSC-631570 (Ukrain) are successfully used for treatment of malignant melanoma since 1996. Melanoma cells of different origin have distinct susceptibility to components of Ukrain. AIM: To carry out comparative investigation of the effect of Ukrain used alone and in combination with pathogen associated molecules (PAM) on mitotic cycle and apoptosis induction in mouse melanoma cell lines with different biological properties. METHODS: Two cell lines with different biological properties (rate of cell division, level of hematogenous metastasis, sensitivity to tumor necrosis factor (TNF)-induced apoptosis) established from B16 mouse melanoma cell line, were used. Apoptosis induction and cell viability were analyzed using trypan blue exclusion test, morphological criteria, DNA gel electrophoresis and flow cytometry. Cell cycle distribution of tumor cells was determined by flow cytometry. Transporters associated with antigen processing (TAP) genes expression was analyzed using reverse transcriptase-polymerase chain reaction (RT-PCR) method. RESULTS: The melanoma cells with different metastatic capabilities differed markedly by the growth rate, sensitivity to apoptosis inducers, and the character of TAP gene expression. Treatment of melanoma cells with Ukrain resulted in apoptosis induction in a dose dependent manner. Melanoma cells with high-metastatic properties were more sensitive to Ukrain than their low metastatic variants. However combined use of drug with PAM induced apoptosis more effectively in melanoma cells with low-metastatic potential. CONCLUSION: Sensitivity to Ukrain in vitro may depend on biological properties of melanoma cells and may be modified by combined treatment of cells with TLR ligands. The results can be useful to optimize the regimen of mono and combined treatment of melanoma with Ukrain.

Comparative investigation of the effect of ukrain on growth of ascites and solid forms of Ehrlich's carcinoma.

UNLABELLED: Ukrain (NSC-631570) is a cytostatic and immunomodulating semisynthetic compound of thiophosphate-modified alkaloids of Chelidonium majus L. It has selective cytotoxicity against cancer cells without healthy cells damaging. Dosage range, methods of introduction and duration of administration of the drug vary. AIM: To carry out comparative investigation of effect of Ukrain on growth of different form of Ehrlich's carcinoma. METHODS: Ehrlich's carcinoma cells were transplanted intraperitoneally and subcutaneously between the scapulas. Ukrain was administered intraperitoneally for 6 days (0.25 mg per mice of 20 g, it's 0.1 LD50). The effect of drug on tumor growth was evaluated by the indexes of tumor growth inhibition (in the case of solid form), total number of tumor cells in ascites, number of viable tumor cells (in the case of ascitic form) and average life span of experimental animals. Cell cycle distribution of cancer cells was determined by flow cytometry. The number of circulating phagocytes was determined by flow cytometry with use of FITC-labelled S. aureus Cowan. RESULTS: Intraperitoneal Ukrain administration in mice with ascite form of Ehrlich's carcinoma resulted in moderate tumor growth retardation, but was accompanied by acute local inflammation and caused reduction of life span of experimental animals. In mice bearing solid form of Ehrlich's carcinoma treatment with Ukrain led to significant tumor growth inhibition and slight increase of life span. In mice bearing both of tumor variants treatment with drug caused restitution of the number of circulating phagocytes in peripheral blood, more expressed in mice bearing ascite tumor variant. CONCLUSION: Thus, anticancer activity of the Ukrain is more expressed in the case of solid variant of Ehrlich's carcinoma. This effect is mediated by direct apoptotic action of drug on Ehrlich's carcinoma cells and positive immunomodulating effect on tumor-bearing organism.

[Tumor-associated macrophages in the prospect of development of targeted cancer therapy].

Tumour-associated macrophages (TAM) make about 80% a total stromal leucocytes in solid tumours. Phenotypic and functional plasticity of this cell population causes its dual effect on tumour grows. Exceptional role of TAM in tumour progression makes them an attractive targets for selective cancer therapy. In this review the main groups of TAM-oriented methods of cancer therapy directed on inhibition of tumour angiogenesis and reactivation of antitumour action of TAM are analysed.

[Responses with participation of toll-like receptors in protective immunity and in pathologic states].

Toll-like receptors (TLRs) are a group of transmembrane receptors which play a key role in both innate and adaptive immune responses. The specific exogenous ligands of TLRs are pathogen-associated molecular patterns such as peptidoglycan, flagellin, teichoic acid, CPG-containing DNA, and others. Stimulation of TLRs induces synthesis and secretion of cytokines, upregulation of co-stimulatory molecules and functional maturation of antigen-presenting cells and leads to the development of both protective and damaging adaptive immune reactions. TLRs are also able to interact with a number of endogenic ligands such as fibronectine, heat shock proteins and extracellular matrix components. Thus, TLRs are involved in the development of many pathological states including sepsis and aseptic inflammation, allergy and autoimmune diseases, cancer. In the recent years several biotechnology and pharmaceutical companies developed new drugs that are either agonists of TLRs to enhance immune responses against tumours and infecious agents or to correct inadequate immune reactions or antagonists designed to reduce the inflammation caused by infection or autoimmune diseases. The paper presents current data concerning TLRs biology, the contribution of TLRs to pathogenesis of human diseases and completed, ongoing and planned clinical trials with immunotherapeutic drugs based on TLR agonists and antagonists.

Influence of teichoic acid from S. aureus on metabolic activity of macrophages and cytotoxic activity of splenocytes of mice bearing Lewis lung carcinoma.

AIM: To investigate the effect of teichoic acid (TA) from the cell wall of S. aureus on some indices of immunological reactivity in mice bearing Lewis lung carcinoma (LLC). METHODS: The teichoic acid at the doses of 1, 2 and 4 microg/g of body weight has been administered subcutaneously simultaneously with tumor cells transplantation and in 7 days. The cytotoxic activity of peritoneal macrophages has been assessed by NBT-test. The splenocyte cytotoxic activity against the LLC cells has been tested by flow cytometry. The evaluation of tumor infiltration by lymphoid cells was carried out as well. RESULTS: TA had no significant effect on oxidative metabolism of peritoneal macrophages in tumor bearing mice. Upon TA administration, the cytotoxic activity of splenocytes against the LLC cells has been augmented in a dose-dependent manner (at the TA dose of 4 microg/g, 2-fold decrease of tumor growth and metastasis has been registered) and leads to decreased tumor infiltration by mononuclear cells. CONCLUSION: TA caused a dose dependent inhibition of growth and metastasis of LLC. It was supposed that TA can influence the tumor grows by activation of splenocytes cytotoxic activity.

Circadian rhythms of cytotoxic activity in peripheral blood mononuclear cells of patients with malignant melanoma.

AIM: To study circadian rhythms (CR) of cytotoxic activity in peripheral blood mononuclear cells of patients with malignant melanoma were compared with those in healthy men. METHODS: The NK-cell and phagocyte cytotoxic activity in five patients with malignant melanoma stage I or II and 12 healthy donors has been assessed by radioimmune assay and NBT-test. RESULTS: The circadian rhythmicity in NK-cells and phagocyte activity in all cancer patients under study has been disrupted. The extent of such disruption tended to increase in patients with more advanced cancer. The most typical alterations were discoordination between the cytotoxicity rhythms of NK-cells and phagocytes (synchronized in healthy persons) and alterations in basic rhythm parameters: phase shifts and amplitude damping. CONCLUSION: In melanoma patients the significant alteration of CR in NK-cells and phagocytes cytotoxic activity was revealed. In spite of individual variations, the degree of the rhythm disruption basically depended on a disease stage. The alteration of CR phase and amplitude and discoordination between the rhythms of NK-cells and phagocyte were registered in all cases studied.

[The effect of electromagnetic radiation with extremely high frequency and low intensity on cytotoxic activity of human natural killer cells]. / O vliianie élektromagnitnogo izlucheniia kraine vysokoi chastoty i nizkoi intensivnotsi na tsitoksicheskuiu aktivnost' estestvennykh killernykh kletok cheloveka.

EHF electromagnetic radiation under short-time action suppresses the cytotoxical activity of the natural killer cells from granulocyte fraction and peripheral blood of healthy volunteers; the observed effect is non-linear. Under the long-time irradiation of the natural killer cells from the mononuclear fraction of blood, the suppressing effect gets a practically linear character after the 20-30 minutes action. Under the long-time irradiation of peripheral blood the insignificant stimulation of natural killers was observed. It is assumed that the radiation applied can suppress the cytotoxic activity of the natural killers, breaking the normal metabolic pathway of phosphatidylinositphosphate.