SYSTEMIC INFLAMMATORY INDICES IN PATIENTS WITH MALIGNANT GLIOMAS AND EFFECTS OF PLATELET SECRETOME IN VITRO.

BACKGROUND: To date, no significant clinical progress has been achieved in the treatment of brain malignant gliomas (MG), and the active search for non-invasive circulating biomarkers continues. The prognostic significance of the ratio of the main peripheral blood cell populations of patients with MG is evaluated. Considerable attention is paid to the secretome of platelets (Pt) of peripheral blood. AIM: To evaluate the indicators of the peripheral blood cell population ratios in patients with brain MG and to study the influence of the secretome of Pt (SPt) of the peripheral blood of patients with brain MG in cell cultures in vitro. MATERIALS AND METHODS: We studied samples of peripheral blood from patients with glioma CNS WHO grade G2 (n = 5), G3 (n = 12), and G4 (n = 20). The peripheral blood cell counts were analyzed in the preoperative period on an automatic hematology analyzer. The in vitro study of SPt was performed on the U251 human glioblastoma cell line cultured with SPt from MG patients or SPt pre-incubated with anti-TGF-ß1 antibody. Cell cultures were observed for 72 h, and mitotic index (MI) was calculated. RESULTS: In MG patients, the count of peripheral blood leukocytes and neutrophils increased (p < 0.05). The neutrophil-to-lymphocyte ratio (NLR) and systemic immune-inflammation index (SII) increased by 2-3 times compared to control. Nevertheless, correlation analysis did not reveal significant relationships between quantitative indicators of peripheral blood cells and the tumor malignancy degree in MG patients. The MI in U251 cells increased under the influence of SPt from patients with MG (p < 0.021), correlated with the tumor degree of malignancy (r = 0.246, p = 0.014). Pre-incubation of SPt with anti-TGF-ß1 antibody tends to neutralize this promitotic effect. CONCLUSION: In MG patients, the integral indicators of NLR and SII increased but no significant relationship with the degree of tumor malignancy was found. In U251 cells, promitotic effects of SPt of MG patients partially decreased by anti-TGF-ß1 antibody.

TGF-ß1 expression by glioma C6 cells in vitro.

The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor ß1 (TGF-ß1) and p53 in C6 cells of rat glioma in vitro. MATERIALS AND METHODS: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-ß1 monoclonal antibody. Immunocytochemical staining for TGF-ß1 and p53 was performed. RESULTS: The proportion of TGF-ß1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-ß1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-ß1 monoclonal antibody, the above effects of FRBCS were abrogated. CONCLUSION: The obtained results suggest that TGF-ß1 seems to be responsible for decrease in TGF-ß1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS.

[THE INFLUENCE OF PROGENITOR NEURO- CELLS SUPERNATANT ON THE LYMPHO- CYTES CYTOTOXIC FUNCTION IN RATS WITH GLIOMA].

The impact of rat neurogenic progenitor cells supernatant (RPNS) on the cytotoxic function of lymphocytes in rats under conditions of physiological norm and experimentally modeled tumor (brain glioma strain 101.8) was studied. The research was carried out in animals with inoculated tumor without RPNS injection and with different regimes of RPNS injection (thrice repeated from 5th to 10th day after glioma inoculation as well as 1 week and 1 month before tumor inoculation). Comparison groups included rats without glioma who triple injected with RPNS; and intact animals (control). RPNS was received from suspension of neurogenic progenitor cells (NPC) of rat brain on 14th day of gestation and injected intraperitoneally (0,12 mg per animal). Cytotoxic function of lymphocytes of experimental rats was evaluated in MTT-colorimetric test with allogeneic glioma cells. RPNS administration increased the cytotoxic activity of lymphocytes in vitro tests with allogeneic tumor cells in intact animals (to 37-38%) as well as in rats with glioma (to 11-22%). Under the RPNS influence the life expectancy and median survival of tumor-bearing animals increased (an average of 3-4 days). RPNS input modes such as triple injection from 5th to 10th day after glioma inoculation and 1 week before inoculation were the most effective. Thus, indirect tumor-inhibiting effect under intraperitoneal. RPNS administration in rats with glioma is demonstrated, which is obviously due to increased efficiency of cytotoxic function of immune cells of animals with inoculated tumor under the influence of the factors produced by NPC.

Anticancer efficacy of allogeneic vaccine modified with progenitor neural cells supernatant in rats with glioma 101.8.

UNLABELLED: The aim of the work was to investigate the antitumor efficacy of allogeneic tumor vaccine (ATV) modified with rat progenitor neural cell supernatant (RPNS) in rats with glioma 101.8. MATERIALS AND METHODS: The study was performed on 74 white random-bred rats. ATV was developed on the basis of glioma 101.8 cell suspension modified with RPNS (0.02 or 0.10 mg/ml). RPNS was prepared from suspension made from whole rat brain tissue on 14(th) (E14) day of gestation. Model of brain glioma 101.8 was reproduced by intracerebral injection of glioma 101.8 cell suspension. ATV was injected intraperitoneally in a volume of 0.2 ml per animal once on the 10(th) day after tumor transplantation. For survival analysis Kaplan - Mayer multiple assessments method was used. Cytotoxic activity of rat lymphocytes (effector cells) was evaluated in MTT-colorimetric test by determining the state of mitochondrial dehydrogenase enzymes in target cells (allogeneic glioma 101.8 cells). RESULTS: Intraperitoneal administration of ATV modified with 0.10 mg/ml RPNS significantly increased mean survival time and median survival of glioma-bearing rats compared with unvaccinated group (MST (19.9 ± 2.4), 21.4 days; versus MST (14.6 ± 2.8); 14 days; p = 0.0002, Gehan's - Wilcoxon test). Intraperitoneal administration of ATV modified with 0.10 mg/ml RPNS resulted in increased cytotoxic activity of immune cells of rats with glioma in vitro compared with this index in unvaccinated group (p = 0.026, U-Mann - Whitney test). CONCLUSION: Antitumor effect of vaccination with RPNS-modified ATV is realized via increased cytotoxic activity of immune cells, what could be used for further optimization of whole tumor cell vaccine.

[Effect of spacer arms on the properties of trypsin and papain immobilized on viscose fibers]. / Vliianie mediatorov na svoistva preparatov tripsina i pepsina, immobilizovannykh na viskoznom volokne.

Properties of the enzymes--trypsin and papain immobilized on viscose fibers were studied, using two spacer arms: maleic dialdehyde and cyanuric chloride. The enzyme thermostability, urea denaturation and resistance to autolysis were examined. Greater denaturation stability was achieved with the use of cyanuric chloride as a spacer arm.