Human Bhsp90 as adjuvant in HCV recombinant vaccine

There are more than 350 million individuals with hepatitis C in the world. One of the important problems in vaccine project is development of effective and suitable adjuvant in human vaccines. At present research we applied human BHsp90 protein as an adjuvant in recombinant HCV vaccine design. The thermal vector of pGP1-2 was used for human heat shock protein 90 expression. This protein injected to BalbC mice as an adjuvant together with recombinant protein of HCV core. The combination of these proteins was used and we evaluated the humoral and cellular immunity and the cytokine secretion of inguinal and popliteal lymph nodes lymphocytes were analyzed in vitro and ex vivo conditions. So the combination of Core protein together with hsp90 induced total IgG and IgG2a secretion. The spleen lymphocytes proliferation were increased equal to serum IgG2a level that was constant in second time bleeding with significant different to complexes with freund's adjuvant. At first IL-4 and IL-5 cytokines were increased, after one week it decreased. Production of IL-4 showed there was no hypersensitivity reaction after vaccine injection

in vitro assessment of antimicrobial and cytotoxic effects of nanosilver

The antimicrobial activity of silver nanoparticles has been investigated in medical fields in recent years, but there are few studies regarding its effect on oral microorganisms. The aim of the present study was to evaluate the in vitro antimicrobial and toxicity properties of nanosilver against two dental plaque microorganisms and Human Gin-gival Fibroblast [HGF] cell line. Antibacterial effects of nanosilver colloidal solution were determined by minimal inhibitory concentration [MIC] and minimal bactericidal concentration [MBC] using microdilution method. Standard strains of Streptococcus sanguis and Actinomyces viscosus were used. For toxicity assessment, MTT and LDH tests were performed under controlled conditions. Different concentrations of nanosilver were prepared and their toxic effects on HGF were determined after 24, 48 and 72 hours. The MIC of nanosilver solution for S. sanguis and A. viscosus were 16 and 4 microg/ml, respectively. The MBC of nanosilver was 64 microg/ml for S. sanguis and 16 microg/ml for A. viscosus. MTT results showed that after 24 hours the concentrations of > 0.5 microg/ml of nanosilver solution affected cell viability when compared with control group. After 48 and 72 hours only the concentration of > 5 microg/ml showed significant effect on cultured cell viability. LDH release test demonstrated toxic effect only after 48, 72 hours by 20 and 50 microg/ml of nanosilver. The results demonstrated that beside its antibacterial activity against S. sanguis and A. viscosus, nanosilver mediated a concentration and time dependent cytotoxicity on HGF

effect of MS14 on production of pro-inflammatory cytokines by macrophages

Using herbal medicines as a complementary treatment method is increasing in wide variety of diseases. MS14-an herbal-marine preparation-is reported to have anti-inflammatory and immunomodulatory activities; however, the mechanism underlying its therapeutic effect is not known. Macrophages play an important role in host defense mechanisms and carry out their role by producing various mediators including proinflammatory cytokines [TNF alpha, IL-1beta]. In this study the effects of orally administered MS14 on TNF alpha and IL-1beta production of BALB/c mice peritoneal macrophages were evaluated. MS14 at 100 mg/kg was orally administered for 5 days to BALB/c mice in MS14 group. Sterile normal saline was administered to mice in control group. Peritoneal macrophage were isolated from control and MS14 groups and cultured, then the supernatants were collected and the cytokines IL-1beta and TNF alpha were measured by ELISA test. Significant decrease in TNF alpha and IL-1beta production of macrophages both at the presence and absence of stimulators was observed. TNF alpha levels were 64.7 +/- 4.6 and 51.1 +/- 4.2 pg/ml in drug and control groups respectively [P< 0.05] and 298.7 +/- 31.3 and 177.0 +/- 26.5 pg/ml in stimulated [PMA+fMLP] cultures of drug and control groups respectively [P< 0.007]. The IL-1beta levels was 130.1 +/- 2.8 pg/ml in control and 65.1 +/- 5.6 in MS14 group [P< 0.000]. It could be concluded that MS14 is able to cause a decline in some inflammatory responses of immune system, which could be considered as at least one of its immunomodulatory mechanisms

[Anti-proliferative and pro-apoptotic effects of fractions from pleurotus florida on HT-29 colon cancer cells]

Edible mushroom, Pleurotus florida [P.florida] has been used by mankind in ancient times because of its nutritional values and medicinal benefits. Cytotoxicity of fractions isolated from P.florida has been reported. The aim of this study was to isolate some fractions from P.florida and evaluate its cytotoxicity effects on colon cancer cells. In this basic study, R5, R10, R30 and R100 fractions were prepared from P.florida and their cytotoxicity activity were evaluated on HT-29 and HGF cell lines. Also, pattern of cell death was determined. Tumoral [HT-29] and non-tumoral [HGF] cells were treated with various concentrations of isolated fractions. MTT assay was used for the evaluation of cell viability. Pattern of cell death was determined using annexin V and propidium iodine staining followed by FACS analyses. Obtained results were analyzed by SPSS software using ANOVA test. R5, R10, R30 and R100 fractions inhibited cell viability of HT-29 cells in a concentration-dependent manner, but had less cytotoxicity on normal fibroblast-like cells [HGF]. Their IC50 values were 46%, 46%, 8% and 4%, respectively. R30 and R100 had the most anti-inhibitory effect. These fractions inhibited cell viability mostly via induction of early apoptosis in colon cancer HT-29 cells at 18%, 49%, 64% and 72%.Our results showed less sensitivity to R5, R10, R30 and R100 fraction in normal cells in comparison to tumoral cells. These fractions also had significant cytotoxic effect on colon cancer cells. Thus, isolated fractions may be considered candidates as chemotherapeutic agents in cancer treatment in future

effect of MS14 on body weight, spleen index and structures of kidney, liver, spleen, lung, lymph node and bone marrow histology in Balb/c mice

MS 14 is an Herbal-marine preparation that has been used in experimental studies for the management of Multiple sclerosis, [MS]. In this study the effect of MS 14 on body weight, spleen index and the histological picture of various organs was evaluated. Female Balb/C mice of 6-8 weeks age were divided into control and test groups. MS 14 was orally administrated at a dose of 100 mg/kg for five days to the experimental group and normal saline given to the control group. After euthanasia on day six, the body weight was measured, spleen index was calculated and representative pieces of tissues including kidney, liver, spleen, lung, lymph node and bone marrow were collected in 10% formalin solution and processed through a standard paraffin embedding method. Sections of 5 micrometer thickness were cut and stained with hematoxylin and eosin. MS 14 at 100 mg/kg did not affect body weight and spleen index, but in the test group, at least 50% of spleen and 90% of lymph node micro sections showed lymphoid hyperplasia: no reactive changes were observed in controls. In both groups, histological evaluation of kidney, liver, spleen, lung, lymph node and bone marrow micro sections showed no significant histological alterations in the normal architecture. According to result of this study, it seems that although MS 14 has no effect on body weight and spleen index, it may induce hyperplastic changes in spleen and lymph nodes, thus signaling activation of the immune system

Substance P potentiates TGF-beta1 production in lung epithelial cell lines

Transforming growth factor-beta [TGF-Beta] is one of the most important cytokines implicated in growth, differentiation, repair and also the pathogenesis of the lung fibrosis by its stimulatory effect on extracellular matrix deposition. Pulmonary epithelial cells are considered as a source of TGF-Beta in lung. Substance P [SP], as a neuroimmunomodulator has elevated levels in inflamed airways and although it has significant role in the pathogenesis of the lung fibrosis, but its effect on transforming growth factor -beta [TGF-Beta] production of the lung epithelial cells [and so its regulatory potential] remains unclear. In this study TGF-Beta1 levels in supernatants of the normal [BEAS-2B] and cancerous [A549] lung epithelial cell line cultures at the presence of various concentrations of SP were examined and MTT assay was performed to evaluate cells viability. We have observed that SP [without any other stimulator] significantly augments TGF-Beta production of both BEAS and A54 cells and this effect is inhibited by NK1-receptor antagonist [CP-96345]. We have also observed that the viability of cells did not significantly affect at the presence of SP. It can be concluded that SP can directly modulate the release of TGF-Beta from human bronchial epithelial cell line and thereby participates in various lung functions or pathologic conditions