Inhibitory effect of prolactin on Toxoplasma proliferation in peripheral blood mononuclear cells from patients with hyperprolactinemia.

Despite many years of studies on the mechanisms of immunological defence responses induced in host organisms by Toxoplasma, no satisfactory immunoprophylaxis or chemotherapy have yet been established for humans. Thus, alternative methods to prevent toxoplasmosis and to enhance the efficacy of currently used antitoxoplasmic drugs are under evaluation. In this work, we strove to determine the influence of human prolactin (endogenous present in serum--sPRL and recombinant--rhPRL) on the course of Toxoplasma infection of peripheral blood mononuclear cells (PBMC) originating from female hyperprolactinemia patients. This study revealed that exogenous rhPRL as well as autologous sPRL from inactivated sera significantly restricted intracellular growth of Toxoplasma in PBMC cultures. Moreover, analysis of IL-10 production by PBMC infected with Toxoplasma and cultured in the presence of sPRL showed a positive correlation between sPRL concentration and the level of IL-10. The obtained results could indicate the possible protective action of PRL in a host organism infected with Toxoplasma and suggest that a significant increase in the serum PRL level, during pregnancy for instance, might significantly limit the risk of Toxoplasma spreading and could play an important role in natural protection against toxoplasmosis. The mechanism of inhibitory effect of PRL needs further detailed study.

Osteopontin and endostatin concentrations in peripheral blood of patients with adrenal tumors undergoing unilateral adrenalectomy.

Peripheral blood osteopontin (OPN) and endostatin (END) levels were studied in 35 patients with adrenal cortex tumors and 10 patients with pheochromocytoma before unilateral adrenalectomy, as well as in 10 healthy subjects (controls). Thirty days after surgery, OPN and END were evaluated again in 16 patients with adrenal cortex tumors and 4 female patients with pheochromocytoma. Before surgery, OPN blood concentrations increased in the group of patients with adrenal cortex carcinomas as compared to controls (p < 0.001) and the group with Conn syndrome (p < 0.05); they did not change after surgery. Before adrenalectomy, OPN blood levels in pheochromocytoma patients were also lower than in Conn syndrome subjects (p < 0.05). After adrenalectomy, the normal concentrations of END decreased only in the group of patients with hormonally inactive cortical adenomas (p < 0.05). We were unable to demonstrate any relationships between removed tumor volumes and OPN or END blood levels.

Resorcylic acid lactone L-783,277 inhibits the growth of the human adrenal cancer cell line H295R in vitro.

The resorcylic acid lactone L-783,277, isolated from a Phoma sp. (ATCC 74403), is a potent and specific inhibitor of MEK (Map kinase kinase) that exerts very interesting pharmacological activities including anti-neoplastic properties. However, the role of this compound in the regulation of endocrine-related cancer cell growth and tumor progression remains unknown. In the present study we have evaluated the effect of L-783,277 on the viability, proliferation and cell cycle of the human adrenocortical carcinoma cell line H295R. L-783,277 inhibited viability (IC50 of 22 microM) and cell proliferation (IC50 of 21 microM) of H295R. At concentrations of 10(-6)-10(-8)M this effect was associated with the accumulation of H295R cells in S-phase, whereas at concentrations of 10(-9)-10(-10)M a prolonged G1-phase and reduced transition into S-phase were observed. Our findings demonstrate for the first time the anti-proliferative action of L-783,277 on the human adrenocortical H295R cell line.

Effects of somatostatin-14 and the receptor-specific somatostatin analogs on chromogranin A and alpha-subunit (alpha-SU) release from "clinically nonfunctioning" pituitary adenoma cells incubated in vitro.

The aim of the study was to examine the effect of somatostatin (SST) and its analogs on the release of chromogranin A (CgA) and alpha-subunit (alpha-SU) from clinically non-functioning pituitary adenomas incubated in vitro. Seven pituitary macroadenomas surgically removed were investigated. All of the tumors were diagnosed before surgery as non-functioning, but they expressed either gonadotropins or their subunits as detected by immunohistochemistry. Two tumors additionally expressed prolactin and growth hormone. All adenomas also expressed chromogranin A (CgA) and at least 3 of 5 subtypes of somatostatin receptors. The cells isolated from the examined tumors were exposed in vitro to either native SST-14 or the following receptor-specific SST analogs: BIM-23926 (agonist of sst1 receptor), BIM-23120 (agonist of sst2 receptor), BIM-23206 (agonist of sst5 receptor) and BIM23A387 (somatostatin/dopamine chimera). The concentration of CgA was measured by means of ELISA method and of alpha-SU was measured by an immunoradiometric method. It was found that the exposure on SST-14 resulted in the decrease of CgA and alpha-SU release from tumor cells in majority of samples, and the effect on CgA was positively correlated with the expression of sst3 and also with the sst2A/sst2B expressions ratio. The inhibitory effect of SST-14 on CgA and alpha-SU seems also to correlate negatively with the expression of sst2B. CgA inhibition also correlates positively with sst5 expression. Among the other compounds studied, only the sst2 agonist decreased the release in all the investigated samples. The remaining substances (agonists of sst1 and sst5 and SST/DA chimera) produced the divergent changes (increased or decreased release, depending on the sample). The data suggest that the inhibition of CgA (and possibly of alpha-SU) release by SST is mediated via subtypes sst2A, sst3 and sst5, whereas sst2B subtype may induce the opposite effect.

Effect of the growth hormone-releasing hormone [GHRH(1-44)NH2] on IL-6 and IL-8 secretion from human peripheral blood mononuclear cells in vitro.

OBJECTIVE: Bidirectional communication between the neuroendocrine and immune systems is now a subject of an intensive investigation. Growth hormone-releasing hormone (GHRH) is synthesized by the hypothalamus, but is present also in the immune cells. Some recent data indicate also an immunomodulatory role of the neuropeptide. The aim of the study was to examine the influence of GHRH(1-44)NH2 on interleukin-6 and interleukin-8 secretion from human peripheral blood mononuclear cells cultured in vitro. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated by centrifugation using Böyum technique and cultured in a humidified atmosphere of 5 % CO2 and 95 % O2 at 37 degrees C for 24 hours in the presence of lipopolysaccharide (LPS) at the concentration of 2 microg/ml and GHRH(1-44)NH2 (the final neuropeptide concentrations to be tested were 10(-12) to 10(-6) M). ELISA methods were used to measure IL-6 and IL-8 concentrations in the supernatants of cultured cells RESULTS: GHRH(1-44)NH2 influenced IL-6 secretion from cultured cells, but significant inhibition of IL-6 release was observed at 10-6 M (p < 0.001). The negative correlation between the GHRH concentration studied and the IL-6 level in the supernatants was found (r = -0.759; p < 0.001). GHRH had no influence on the secretion of IL-8 from activated PBMC. CONCLUSIONS: Our results demonstrate that GHRH in vitro modulates IL-6 secretion from the human peripheral blood mononuclear cells, without any significant effect on IL-8 secretion.

Effect of growth hormone-releasing hormone (GHRH) and GHRH antagonist (MZ-4-71) on interferon-gamma secretion from human peripheral blood mononuclear cells in vitro.

Numerous reports indicate close interactions between the neuroendocrine and the immune systems. Hypothalamic neuropeptide, growth hormone-releasing hormone (GHRH) stimulates growth hormone (GH) secretion from the anterior pituitary gland, but recently some immunomodulatory properties of this peptide have also been demonstrated. In the present studies we evaluated the effect of human synthetic GHRH(1-44)NH(2) and GHRH antagonist (MZ-4-71) on interferon (IFN)-gamma secretion from human peripheral blood mononuclear cells (PBMC). GHRH(1-44)NH(2) at 10(-10), 10(-8) and 10(-6) M concentrations significantly (p < 0.05) increased the IFN-gamma level in supernatants of cultured cells, as compared with the controls. GHRH antagonist (MZ-4-71) at 10(-10), 10(-8) and 10(-6) M concentrations diminished the IFN-gamma level in supernatants in a dose-dependent manner, but statistically significant differences were observed only at 10(-8) M and 10(-6) M (p < 0.05 vs controls). Our results demonstrate that GHRH and GHRH antagonist MZ-4-71 can modulate IFN-gamma secretion in vitro by human peripheral blood mononuclear cells.

GH-RH antagonist (MZ-4-71) inhibits VEGF secretion and proliferation of murine endothelial cells.

Angiogenesis plays a key role in solid tumor formation, invasiveness and metastasis. Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that is necessary in the process of neovascularisation. Antagonists of growth hormone-releasing hormone (GH-RH) have been shown to suppress both in vivo and in vitro growth and metastasis of many human cancer cell lines. The mechanisms that mediate the antitumorigenic actions of these antagonists involve direct and indirect pathways, but are not completely elucidated. We have examined the effect of GH-RH antagonist MZ-4-71 on proliferation activity and VEGF release from cultured murine endothelial cells HECa10 in vitro. MZ-4-71 at 10(-8) to 10(-6) M concentrations inhibited the proliferative activity of cultured cells and suppressed the release of VEGF into supernatants of 72 h endothelial cell cultures. To our knowledge this is the first study reporting antiangiogenic properties of GH-RH antagonists.

Effect of somatostatin and octreotide on proliferation and vascular endothelial growth factor secretion from murine endothelial cell line (HECa10) culture.

Angiogenesis, development of new blood vessels, is required for normal tissue repair and also for tumor cell proliferation, extracellular matrix invasion, and hematogenous metastases. Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that has been shown to play a key role in neovascularization. Inhibition of angiogenesis in vitro and in vivo was documented by administration of native neuropeptide somatostatin and its analog octreotide. We have studied the effect of somatostatin-14 (SRIF) and ocreotide (sandostatin) on proliferation activity and VEGF release from cultured murine endothelial cells HECa10 in vitro. SRIF in concentrations from 10(-9) to 10(-5) M and ocreotide in concentrations from 10(-9) to 10(-5) M diminished the proliferative activity of cultured cells vs controls. SRIF and ocreotide in concentrations from 10(-14) to 10(-6) M did not change the release of VEGF into supernatants of 24 or 72 h endothelial cell cultures. Although we showed the antiproliferative effect of SRIF and ocreotide on mouse endothelial cells, we were unable to demonstrate the inhibitory effect of tested peptides on VEGF secretion in vitro.