Olive (Olea europaea) leaf extract alters the cytokine profile of Leishmania major-infected macrophages: New insight into the underlying mechanism.

This study aimed to identify the effects of olive leaf extract (OLE) on IFNγ, TNFα, TGFß and nitric oxide (NO) resulted from macrophages infected with Leishmania major (L. major) amastigotes in the culture medium. High-performance liquid chromatography (HPLC) was used to analyse the level of Oleuropein in plant extract. To evaluate the immunomodulatory effects of OLE, the isolated BALB/c mice peritoneal macrophages were infected with L. major promastigotes and treated with 6.25, 12.5 and 25 µg/mL concentrations of OLE. To assess the cytokines, supernatants of cell cultures were harvested after 12, 24 and 48 hours. Cytokine production was evaluated by ELISA. Nitrite accumulation in the culture medium was assessed using the Griess reaction. The level of Oleuropein in the extract was 18.45% by HPLC. According to results, the production of IFNγ and TNFα was significantly increased when the infected and/or not infected macrophages with L. major promastigotes were affected by different concentrations of OLE. Conversely, the production of TGFß was significantly decreased under the same conditions. Furthermore, the colorimetric determination of NO accumulation in the culture medium indicated that OLE has no effect on NO production. The study corroborates the immunomodulatory effects of OLE on L. major-infected macrophages.

Vaccination with human amniotic epithelial cells confer effective protection in a murine model of Colon adenocarcinoma.

As a prophylactic cancer vaccine, human amniotic membrane epithelial cells (hAECs) conferred effective protection in a murine model of colon cancer. The immunized mice mounted strong cross-protective CTL and antibody responses. Tumor burden was significantly reduced in tumor-bearing mice after immunization with hAECs. Placental cancer immunotherapy could be a promising approach for primary prevention of cancer. In spite of being the star of therapeutic strategies for cancer treatment, the results of immunotherapeutic approaches are still far from expectations. In this regard, primary prevention of cancer using prophylactic cancer vaccines has gained considerable attention. The immunologic similarities between cancer development and placentation have helped researchers to unravel molecular mechanisms responsible for carcinogenesis and to take advantage of stem cells from reproductive organs to elicit robust anti-cancer immune responses. Here, we showed that vaccination of mice with human amniotic membrane epithelial cells (hAECs) conferred effective protection against colon cancer and led to expansion of systemic and splenic cytotoxic T cell population and induction of cross-protective cytotoxic responses against tumor cells. Vaccinated mice mounted tumor-specific Th1 responses and produced cross-reactive antibodies against cell surface markers of cancer cells. Tumor burden was also significantly reduced in tumor-bearing mice immunized with hAECs. Our findings pave the way for potential future application of hAECs as an effective prophylactic cancer vaccine.

Effects of hormone replacement therapy on immunological factors in the postmenopausal period.

BACKGROUND: Despite valuable evidence documented on immunological changes in postmenopausal women, particularly following hormone replacement therapy (HRT), it is difficult to explain whether immunological changes during menopause are caused by HRT. This systematic review aimed to summarize the results of studies available on postmenopausal immunological changes and to determine any potential effects of HRT on the immunological profile of postmenopausal women. METHODS: For this systematic review, we primarily explored 751 papers about the immune system status of postmenopausal women published during 1955-2015. Scientific databases including Web of Science, MEDLINE, Scopus, Embase, Google Scholar, and the Cochrane database were searched for a number of relevant key terms. Of 209 papers that met the initial search criteria, 13 papers were potentially retrievable and included descriptions of changes in immunological factors during the postmenopausal period and the effects of HRT on such changes. RESULTS: HRT resulted in a range of immunological changes in postmenopausal women. These changes included reductions in interleukin-2 (IL-2), IL-6, and insulin-like growth factor-1 levels and increments in IL-1 and IL-4 levels. Elevations in B-cell production and estrogen receptor alpha, CD19+ cells, and C3 and C4 complement levels were also documented. Decreased CD8+ counts were also a constant finding in most reviewed papers. However, data on the changes in other factors such as tumor necrosis factor-alpha, interferon-gamma, CD4+, and CD25+ were contradictory. Levels of some immunological factors, e.g. immunoglobulin G (IgG), IgM, and IL-10, remained unchanged following HRT. CONCLUSION: Postmenopausal women are prone to impaired immune responses. HRT during the menopausal period can mediate immunological responses by inducing significant changes in immunological mediators.

Evaluation of immunological interaction between spermatozoa and fallopian tube epithelial cells.

Toll-like receptors (TLR) are one of the major compartments of innate immune system. It was revealed that the TLR have relevance in ovulation, sperm capacitation and fertilisation. So, in this study, the expression of TLR, their adaptor molecules and cytokines in human fallopian tube cell line under the effect of human normal spermatozoa was evaluated. TLR mRNA and protein were evaluated in OE-E6/E7 cell line. Semen samples from 10 donors were collected and co-incubated with OE-E6/E7 cell line and used as sperm group, and cell line without spermatozoa was used as control group. Afterwards, the level of TLR, their adaptor molecule and cytokine mRNA expression was compared using qPCR in sperm and control groups, and supernatant was used for ELISA. To determine whether elevated cytokine reaction to spermatozoa in OE-E6/E7 cell line is mediated via TLR, TLR3 function-blocking antibody was used. OE-E6/E7 cell line expressed TLR1-6 genes and proteins. TLR expressions, especially TLR3 and TLR5, in OE-E6/E7 cell line under the effect of spermatozoa were significantly higher. Also, levels of adaptor molecules and cytokine production were increased in sperm group than in control group (P < 0.05). So, it may be hypothesised that TLR are essential for spermatozoa and fallopian tube immunological interaction and for preparing safe environment for important events in fallopian tube.

Gene regulation of pteridine reductase 1 in leishmania promastigotes and amastigotes using a full-length antisense construct.

BACKGROUND: Pteridine metabolic pathway is unusual features of Leishmania, which is necessary for the growth of parasite. Leishmania has evolved a complex and versatile pteridine salvage network which has the ability of scavenging a wide area of the conjugated and unconjugated pteridines especially folate and biopterin. In this study, we focus on the inhibition of ptr1 gene expression. METHODS: L. major ptr1 gene was cloned into pcDNA3 and digested using KpnI and BamHI. The gene was subcloned so that antisense will transcribe and called pcDNA-rPTR. Leishmania major was cultured and late logarithmic-phase promastigotes were harvested. The promastigotes were divided into two groups. One group was transfected with 50 µg of pcDNA-rPTR, whereas the other group was transfected with pcDNA3. Transfected cells were cultured and plated onto semi-solid media. Mouse pritonean macrophages were transfected using pcDNA-rPTR-tansfected promastigotes. Western blotting was performed on mouse transfected pritonean macrophages and extracts from transfected promastigotes of L. major using a L. major ptr1 antibody raised in rabbits. RESULTS: The PTR1 protein was not expressed in pcDNA-rPTR- tansfected promastigotes and mouse macrophage transfected with pcDNA-rPTR- tansfected promastigotes. CONCLUSION: This approach might be used to study the pteridine salvage pathway in Leishmania or to assess the possibility of using gene expression inhibition in the treatment of leishmaniasis.

An efficient procedure for purification of recombinant human ß heat shock protein 90.

BACKGROUND AND THE PURPOSE OF THE STUDY: Heat Shock Protein 90 (Hsp90) is typically the most abundant chaperone in the eukaryotic cell cytoplasm, and its expression is essential for loading immunogenic peptides onto major histocompatibility complex molecules for presentation to T-cells. Therefore, it may act as a good candidate as an adjuvant molecule in vaccine technology. METHODS: Initially the human Hsp90ß gene was cloned into the heat inducible expression vector pGP1-2 and then the recombinant protein was isolated by ion exchange chromatography. After intradermal injection of confirmed purified band of protein to rabbits and isolation of the serum IgG antibody, for its affinity purification, the rabbit's purified Hsp90 specific IgG was coupled to the cyanogen bromide-activated Sepharose 4B. RESULTS: The recovery of the purified protein of interest by affinity chromatography was 50%. CONCLUSION: This research enabled purification of human heat shock protein by a laboratory prepared column chromatography.

Seroprevalence of Acanthamoeba Antibodies in Rheumatoid Arthritis Patients by IFAT, Tehran, Iran 2007.

BACKGROUND: This preliminary study was conducted to discriminate the prevalence of Acanthamoeba antibodies in rheumatoid arthritis (RA) patients and healthy controls to analyze the correlation between these two groups. METHODS: From October 2006 to August 2007 a total of 121 serum samples from RA patients attending the Rheumatolgy Department at Shariati Hospital in Tehran were obtained and stored at -20°C until using by indirect fluorescent-antibody test (IFAT). RA was diagnosed according to the American Collage of Rheumatology classification criteria. The organism used in this study was isolated from various water resources in Tehran, Iran cultured axenically and then went on a PCR assay based on 18S rRNA to identify the genus Acanthomoeba. Indirect immunofluorescence antibody (IFA) staining of serum samples was carried out to detect anti Acanthomoeba antibodies. RESULTS: In culture, out of 22 samples, 13(59%) were grown in xenic but only two in axenic medium. PCR amplified a 904bp fragment, specific for Acanthamoeba. Of examined serum samples, Acanthamoeba antibodies were present in 70 (57.8%) and 52 (41.2%), respectively. The highest titer of antibodies (1:320) was detected in one patient with RA. CONCLUSION: Our study supports the hypothesis that some parasitic microorganisms can involve and contribute toward the development of rheumatoid syndromes.

Polymorphonuclear leucocyte respiratory burst activity correlates with serum zinc level in type 2 diabetic patients with foot ulcers.

Patients with diabetes mellitus (DM) are prone to infection, in part due to phagocyte dysfunction and impaired polymorphonuclear (PMN) leucocyte superoxide generation. Another frequently mentioned factor in the pathogenesis of infection in DM patients is altered zinc status. This study aims to evaluate the association between serum zinc level and PMN respiratory burst activity in patients with type 2 DM. Thirty-nine type 2 DM patients (19 with foot ulcers) and 20 healthy controls are studied. Respiratory burst activity is evaluated at baseline and in stimulated states by a nitro blue tetrazolium (NBT) reduction test. Serum zinc level is evaluated by atomic absorption spectrophotometry. Although not statistically significant, PMNs from diabetics with foot ulcers appeared to be slightly hyperactivated at the baseline state. The NBT index was significantly lower in DM patients with foot ulcers after stimulation. Mean serum zinc level was significantly lower in diabetics with foot ulcers compared to those without foot ulcers. A significant negative correlation between serum zinc level and NBT index at baseline was seen in patients with foot ulcers, but this changed to a significant positive correlation after stimulation. These findings may be explained by PMN hyperactivity at baseline and by respiratory burst dysfunction following stimulation in diabetic patients.

The immunostatus of T helper and T cytotoxic cells in the patients ten years after exposure to sulfur mustard.

The treatment and cure of patients exposed to sulfur mustard is a remaining challenge despite ongoing research in this field. A severe suppression of the immune system still remains the major cause of opportunistic infections, septicemia, and death in these patients. The patients evaluated in this study were classified into three groups; severe, moderate, and mild according to the severity of exposure to sulfur mustard. We measured the total leukocyte and the status of T helper and T cytotoxic cells in such patients 10 year after being exposed to sulfur mustard. The total leukocyte was measured by using anti CD45+ and the amount of T helper and T cytotoxic were evaluated by anti CD3, anti CD4, and anti CD8, while the activity of T helper was measured by anti CD4 and anti CD25. T helper and T cytotoxic cells were evaluated in lymphocyte and leukocyte gates through forward scatter and side scatter flow cytometer. The results showed that the percentage of CD45+ were normal in all of the groups while the percentage of T helper and T cytotoxic cells were significantly (P < 0.05) decreased in the severe group comparing to mild group. Also the results indicate that CD4+/CD25+ cells in the most severely affected patients were significantly (P < 0.05) increased comparing to the other groups. Ten years following exposure to sulfur mustard, the immune system of the patients is still impaired and this might be related to their present health problems.