[Show abstract][Hide abstract] ABSTRACT: The most important long-term morbidity problem of sulfur mustard (SM) toxicity is pulmonary complications but the pathogenesis of these complications is not clearly understood. This study evaluates the peripheral blood mononuclear sub-sets and their correlation with pulmonary function in SM exposed civilian cases 20years post-exposure as gathered in the context of the Sardasht-Iran Cohort Study (SICS). Samples were randomly selected from two groups, SM-exposed (n=372) and control (n=128), with the same ethnicity, culture, and demography. Three color flow cytometry was applied for peripheral blood mononuclear sub-population determination. Results indicated a significant decrease in CD45+/CD3+, CD45+/CD3+/CD4+, and an increase in CD3+/CD16+56+ percentages. It was also found that absolute count of NK cells was highly increased in peripheral blood of exposed cases. There was a significant increase in NK cell count of SM exposed group with pulmonary problems as compared to the same group without pulmonary problems (p-value<0.04) based on the Global Initiative for Chronic Obstructive Lung Disease (GOLD). The findings showed a significant negative correlation between absolute numbers of T lymphocyte and FVC % and positive correlation with FEV1/FVC%. The results also demonstrated that absolute numbers of monocytes had a negative correlation with FVC %. We propose that NK and T cells are probably involved in the pathogenesis or immune reactions to the delayed pulmonary complications induced by SM. This hypothesis should be tested in a more severe pulmonary complicated group.
International immunopharmacology 02/2013; · 2.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The serum levels of four important and well characterized inflammatory chemokines including MCP-1/CCL2, RANTES/CCL5, IL-8/CXCL8 and Fractalkine/CX3CL1 were evaluated in sulfur mustard (SM) exposed Iranian population 20 years after exposure. In this historical cohort study 372 SM exposed participants from Sardasht, and 128 unexposed participants as controls were studied. The serum concentrations of chemokines were measured by a sandwich ELISA technique. The serum concentrations in the exposed comparing to the control group were 201.86 vs 180.60 pg/ml (p=0.002), for MCP-1/CCL2, 1182.6 vs 1393.1pg/ml (p=0.021) for RANTES/CCL5, 12.61 vs 15 pg/ml (p=0.002) for IL-8/CXCL8 and 0.696 vs 0.0648 (p=0.413) for Fractalkine/CX3CL1. In conclusion, elevated levels of MCP-1/CCL2 may suggest an anti inflammatory response and decreased levels of IL-8/CXCL8 and RANTES/CCL5 may represent a different pathophysiology and diverse molecular mechanisms involved in long term clinical manifestations of SM exposure. However, further prospect into their role in the pathogenesis of SM remains to be done.
International immunopharmacology 10/2009; 9(13-14):1471-6. · 2.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Sulfur mustard (SM) has short- and long-term toxicity against various organs including the respiratory system. However, the basic and molecular mechanisms of SM long-term toxicity have not clearly been defined. Thus, the aim of this study was to evaluate the association of soluble Fas ligand (sFasL) as well as nitric oxide (NO) serum levels with long-term pulmonary complications in a SM exposed population 20 years after SM exposure. In this historical cohort study 372 male SM exposed subjects and 128 age-matched unexposed controls were studied. Clinical evaluation and pulmonary function tests were carried out for all participants and serum concentrations of sFasL and NO measured. According to our results, the serum levels of sFasL and NO were not significantly different between the exposed and control groups. However, the serum levels of sFasL in the exposed group with pulmonary problems were significantly higher than their corresponding in the control group (116.711+/-81.166 vs 86.027+/-30.199 and p=0.028). Furthermore a significant elevation in sFasL levels was found in the exposed subjects with pulmonary problems compared to those exposed participants without pulmonary problems (116.711+/-81.166 vs 90.692+/-57.853 and p=0.004). Based on Global Initiative for chronic Obstructive Lung Disease (GOLD) classification analysis a positive correlation was observed between sFasL levels and pulmonary problems. There was also a significant negative correlation between sFasL and the white blood cell (WBC) count in the SM exposed cohort, but not in the control group. No significant association was shown between NO and pulmonary impairment in the SM exposed subjects. Thus, our results indicate that elevated serum levels of sFasL may be associated with progression of pulmonary diseases in the SM exposed subjects.
International immunopharmacology 10/2009; 9(13-14):1489-93. · 2.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Mustard gas, even in low doses, has the ability to inflict damage in multiple organs especially the skin, eyes, as well as the respiratory tract. This damage may cause many complications which persist during the lifespan of exposed subjects. Pro-inflammatory cytokines including TNF, IL-1alpha, IL-1beta and IL-1Ra cause systemic inflammatory reactions and numerous changes including altered cell signaling and migration, changes in cytokine production and fever. The aim of this study was to determine the serum levels of these cytokines in subjects who were exposed to mustard gas 20 years ago in comparison with an unexposed control group. In this historical cohort study 368 sulfur mustard (SM) exposed participants from Sardasht and 126 age-matched unexposed volunteers from Rabat (a nearby town) as controls were chosen by a random systematic sampling. The serum concentrations of IL-1alpha, IL-1beta, IL-1Ra and TNF were measured by a sandwich ELISA technique. Median of the serum levels of cytokines TNF, IL-1alpha, IL-1beta and IL-1Ra in the control group was 23.79, 1.89, 1.91 and 32.9 pg/ml respectively, while in the SM-exposed participants these values were 11.11, 0.81, 1.73 and 26.7 pg/ml respectively. The serum pro-inflammatory cytokine levels were significantly lower in the exposed group than in controls (p<0.01). There was also significant positive correlation between concentration of all of mentioned cytokines, the strongest being between IL-1beta and TNF (r=0.809 in the control group). The observed down-regulation of pro-inflammatory cytokines should be considered in interpretation of diagnosis and therapeutic measures taken to improve clinical complications.
International immunopharmacology 09/2009; 9(13-14):1466-70. · 2.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Sulfur mustard (SM) is a blistering chemical agent which has short and long term toxicity against many organs. The respiratory tract is one of the main targets, and is the most disabling long term complication of SM. Inflammatory mediators especially IL-8 and IL-6 play the primary role in the various chronic pulmonary diseases. Sardasht-Iran Cohort Study (SICS) was designed to evaluate immunological and molecular parameters in SM exposed people 20 years after exposure. In the present study, the association of the serum levels of IL-8, IL-6, C reactive protein (CRP) and rheumatoid factor (RF) with long term pulmonary involvement was evaluated. There were 348 exposed and 120 control participants. The clinical evaluations were done for all subjects and Spirometry was performed according to American Thoracic Society Criteria. Severity of pulmonary involvement was assessed by Global Initiative for chronic Obstructive Lung Disease (GOLD) classification. The serum levels of IL-8, IL-6 and RF were assessed by ELISA assay. CRP was assessed by photometric method. The serum levels of IL-8 and IL-6 significantly decreased in the SM exposed participants compared to the control group. There were no significant associations between the serum levels of IL-8 and pulmonary symptoms (chronic cough, sputum, hemoptysis, and dyspnea), pulmonary findings (crackles, rales, and wheezing) as well as spirometry parameters. IL-6 was associated with wheezing and CRP was associated with wheezing and rales in SM exposed group. We concluded the serum levels of these inflammatory mediators probably do not have any major role in pathogenesis and persistence of pulmonary complications and do not reflect the degree of severity of pulmonary involvement following SM exposure.
International immunopharmacology 09/2009; 9(13-14):1482-8. · 2.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Lipophosphoglycan (LPG) is structurally characterized by a series of phosphoglycan repeat units. Cellular LPG, isolated from promastigotes, has a very similar structure to culture supernatant LPG, but differs in the average number of phosphorylated oligosaccharide repeat units and in glycan composition. Comparison of these LPGs with capillary electrophoresis and immunoblotting indicate that these molecules are highly conserved structurally and composed of galactosylated Gal-Man repeats but their size and molecular weight are very different which is due to glycan portion. There are 30 and 20 repeat units in sLPG and mLPG, respectively. Both LPGs induced nitric oxide in macrophages cell line while sLPG had the higher stimulatory effect. In the presence of anti-TLR2 nitric oxide stimulated by LPG was reduced to control levels. In addition, in the presence of anti-TLR4, nitric oxide stimulated by LPGs was not affected. We propose that lipophosphoglycan induces nitric oxide production via TLR2 signaling pathway.
[Show abstract][Hide abstract] ABSTRACT: Brucellosis is an infectious disease with high impact on innate immune responses which is induced partly by its DNA. In the present study the potential differences of wild type and patients isolates versus attenuated vaccine strains in terms of cytokines, ROS and NO induction on murine splenocytes and peritoneal macrophages were investigated. This panel varied in base composition and included DNA from B. abortus, B. melitensis, B.abortus strain S19 and melitensis strain Rev1, as attenuated live vaccine. Also we included Escherichia coli DNA, calf thymus DNA (a mammalian DNA), as controls. These DNA were evaluated for their ability to stimulate IL-12, TNF-alpha, IL-10, IFN-gamma and ROS production from spleenocytes as well as NO production from peritoneal macrophages. Spleen cells were cultured in 24 well at a concentration of 106 cells/ ml with subsequent addition of 10 microg/ml of Brucella or Ecoli DNAs. These cultures were incubated at 37 degrees C with 5% CO2 for 5 days. Supernatants were harvested and cytokines, ROS and NOx were evaluated. It was observed that TNF-alpha was induced in days 1,3,5 by all Brucella strains DNAs and E. coli DNA, IL-10 only was induced in day 1, IFN- gamma was induced only in day 5 and IL-12 not induced. ROS and NOx were produced by all strains; however, we observed higher production of NOx which were stimulated by DNA of B. melitensis.
Iranian journal of allergy, asthma, and immunology 09/2009; 8(3):127-34. · 0.65 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the present study, we show for the first time that lipophosphoglycan (LPG) stimulated cytokine production by human peripheral blood mononuclear cells is also mediated via Toll-like receptor (TLR2). In addition, in order to verify if TLR2 is involved in recognition of the purified PGs, neutralizing mAbs against TLR2 and TLR4 were used to treat the cells before being stimulated with PGs. We found strong Th1-promoting cytokines induced by sLPG but not by mLPG which was blocked by presence of anti-TLR2 mAb. This finding reveals a mechanism by which the first encounter and recognition of L. major promastigotes by mLPG after interaction with TLR2 provides a cytokine milieu for consequent Th2 differentiation. Moreover, having shown the strong induction of Th1-promoting cytokines and low production of IL-10 in response to sLPG might have vaccine implication since it is recognized by TLR2 providing signals to professional antigen presenting cells that reside in the skin to promote effective T cell responses against Leishmania infection. In addition, it was shown that purified mLPG and sLPG activate reactive oxygen species (ROS) production which is also blocked by anti-TLR2 but not by anti-TLR4. However, no inhibition was seen in PPG-induced cytokine and ROS production in the presence of anti-TLR2 and anti-TLR4, implying involvement of other receptors.
[Show abstract][Hide abstract] ABSTRACT: Until recently, the only tool for detection of latent tuberculosis infection (LTBI) was the tuberculin skin test (TST). QuantiFERON-TB Gold In-Tube test (QFT) is a promising in vitro diagnostic test for LTBI that has potential advantages over the TST. In this study we aimed to compare QFT with TST for diagnosis of LTBI.
A total of 186 BCG-vaccinated subjects enrolled in study. They underwent TST and QFT assay. They divided in two groups. Group 1 includes individuals who were at low risk for exposure to M. tuberculosis (LRG) and Group 2 includes individuals who were likely to have been exposed to M. tuberculosis infections (HRG).
Overall agreement between QFT and TST was 89.3% (kappa = 0.052). In LRG, agreement between the two tests was 52.6% (95% confidence interval, 44-60%) with kappa-values of 0.019. In HRG agreement between the two tests was 63.2% (95% confidence interval, 42-84%) with kappa-values of 0.28.
In conclusion, the QFT assay showed acceptable results for determining latent M. tuberculosis infection in vaccinated population. The decision to select QFT over TST will depend on the population, purpose of testing and resource availability.
Journal of Evaluation in Clinical Practice 03/2009; 15(1):148-51. · 1.51 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Insights into long-term clinical consequences of sulfur mustard have emerged from some investigations but less is known about the basic and molecular mechanisms of these complications. Sardasht-Iran Cohort Study is a comprehensive historical cohort study on Sardasht chemical victims' population which was designed to find out the long-term complications of sulfur mustard exposure and the basic mechanisms underlying clinical manifestations. This paper describes the design and methodology of Sardasht-Iran Cohort Study.
In Sardasht-Iran Cohort Study, 500 individuals including 372 subjects from Sardasht, as the exposed group, and 128 subjects from Rabat, as the unexposed age-matched control group were evaluated. The exposed group was divided into two groups based on the severity of clinical complications at the time of exposure. Different samples including blood, sputum, saliva, tear, urine, and semen were collected for immunologic, hematologic, biochemical, and other laboratory analysis. Data were gathered from medical records, clinical examinations, laboratory tests, and questionnaires for psychological and lifestyle situations.
The important distinctions setting this study apart from the previous ones are discussed. The Sardasht-Iran Cohort Study provides important information on various aspects of long-term consequences of sulfur mustard exposure. This database will provide a better position to suggest guidelines for the diagnosis, treatment, and prevention of delayed complications in the patients exposed to sulfur mustard.
Archives of Iranian medicine 02/2009; 12(1):5-14. · 1.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: With the aim of determining selectivity and the possible target(s) of nitroheteroaryl-1,3,4-thiadiazoles considered as possible leads for the development of anti-leishmanial agents, we studied 5-nitroimidazole, 5-nitrofuran and 5-nitrothiophene analogs of N-substituted-piperazinyl-1,3,4-thiadiazoles. We investigated 21 representative compounds 1-3(a-g) for the following properties: selectivity and efficiency against different Leishmania wild type species and intracellular parasite, toxicity against host cells and inhibition of topoisomerases I and II. Our results indicate that the nitroimidazole analogs 1a and 1f, and nitrofuran derivatives 2a, 2b, 2c, 2f, and 2g exhibited low toxicity against the host cells (IC(50)> or =80 microM), but high selectivity against intracellular amastigotes (selectivity index>12). Leishmania topoisomerases revealed impressive sensitivity to the agents (%inhibition >50 at IC(50) doses of compounds against Leishmania). Our findings showed that at least part of leishmaniacidal effect of the compounds could be attributed to disruption DNA-relaxed activities of topoisomerases I and II, and cleavable-complex formation.
[Show abstract][Hide abstract] ABSTRACT: Protozoan parasites of the genus Leishmania secrete a range of proteophosphoglycans (PPG) known to be important for successful colonization of Leishmania in the sandfly and for virulence in the mammalian host. PPGs are a large family of extensively glycosylated proteins with some unusual and unique features. In this study we purified PPG from culture supernatant of Leishmania major metacyclic promastigotes. In discontinuous SDS-PAGE, PPG could not enter the resolving gel but after mild acid hydrolysis several bands resolved. Agarose gel electrophoresis and immunoblot analysis using monoclonal antibody (WIC 79.3) indicated that the PPG preparation consisted of heterogeneous molecules. Compositional analysis showed that the PPG preparation contained 67% glycan, 28% protein and 5% phosphate. Additionally, the effect of PPG on reactive oxygen species (ROS) production and induction of IL-10, IL-12 and IFN-gamma secretion by human peripheral blood mononuclear cells (PBMCs) isolated from healthy individuals was investigated. The water-soluble secreted form of PPG at a concentration of 1 microg glycan/ml seems to be a potent inducer of ROS and IL-10 and to a lesser extent of IFN-gamma and IL-12. Cytokines and ROS production was decreased in a dose-dependent manner as the concentration of PPG was increased to 100 microg glycan/ml.
[Show abstract][Hide abstract] ABSTRACT: In spite of reports on cytokines induction by the Brucella DNA in murine model, there is no comparison between pathogenic and appropriate vaccine strains in human. We investigated the cytokines profile of human peripheral blood mononuclear cells (PBMCs) induced by DNA extracted from pathogenic isolates of Brucella melitensis and B. abortus as well as Rev1 and S19; the appropriate vaccine strains. It was observed that despite differential induction of Interleukin(IL)-12 and IL-10 production, identical IL-12/IL-10 concentration ratio was obtained by all Brucella strains DNAs that was 2 after 24 h and 4 after 5 days of incubation. In addition, IL-2 and Interferon(IFN)-gamma production were profoundly increased compared to the medium at day 3 and 5 respectively but IFN-alpha was not induced. Therefore, Brucella strains DNAs are Th1 inducing component with similar pattern in human PBMCs.
Microbiological Research 02/2008; 163(4):466-72. · 1.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Calprotectin is cytotoxic agent that its anti-tumor effects are governed through suppression of topoisomerase II; a key enzyme in apoptosis. In previous studies, cytotoxicity and apoptotic effects of calprotectin are shown on different cancer cell lines, but not human gastric cancer cell lines. In the present study, cytotoxicity and apoptotic effects of calprotectin on human gastric adenocarcinoma cancer cell line (AGS) were evaluated.
The AGS cells were exposed to the different concentrations of calprotectin for 24, 48 and 72 hours. Cell proliferation was assessed using dimethylthiazol diphenyl tetrazolium bromide assay and dye exclusion tests. For evaluation of cytotoxic mechanism in calprotectin on AGS cells, flow cytometric analysis was performed.
Our results revealed that calprotectin induces growth inhibition of AGS in a dose- and time-dependent manner. Results of this investigation showed that sensitivity of AGS cells to cytotoxic effect of human calprotectin was highly remarkable. In addition, growth inhibitory effect of this cytotoxic agent mostly was governed through induction of apoptosis in the AGS cells.
These findings indicated that calprotectin induces growth inhibition and apoptosis in the AGS cells.
[Show abstract][Hide abstract] ABSTRACT: The aim of the present study was to investigate whether induction of differentiation by hyperthermia is accompanied by apoptosis and necrosis to further evaluate the benefits of using hyperthermia as a differentiation inducing physical modality.
Differentiation was evaluated in K562 erythroleukaemia cells by measuring haemoglobin synthesis and flow cytometric measurement of glycophorin A expression. Apoptosis was measured by Annexin-V-FITC and Propidium Iodide (PI) double staining assay. Apoptosis and necrosis was also evaluated morphologically using staining with acridine orange/ethidium bromide (AO/EtBr) by fluorescence microscopy. Heat shock protein 70 (HSP70) level was measured by ELISA kit.
Hyperthermia (43 degrees C) induced differentiation as judged by increased haemoglobin synthesis and glycophorin A expression. No sign of apoptosis or necrosis could be detected at this temperature. Cell viability did not change due to heat treatment, and cellular proliferation was reduced in a dose (heating time) dependent manner. At 45 degrees C, hyperthermia induced apoptosis and necrosis with minimal or no sign of differentiation. HSP70 level was significantly increased at 43 degrees C along with differentiation of leukaemic cells, while at 45 degrees C no significant effect on HSP70 production could be observed.
The encouraging results obtained here indicate that by heat treatment at 43 degrees C, hyperthermia can be used alone or in combination with other modalities as a differentiation inducing agent without any detectable apoptotic activity. Positive correlation between HSP70 production and induction of differentiation and lack of apoptosis by hyperthermia confirm the possible role of HSP70 in the heat-induced differentiation and apoptosis in leukaemic cells.
International Journal of Hyperthermia 01/2008; 23(8):645-55. · 2.59 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Angiocardiography is an X-ray examination of the blood vessels or chambers of the heart. Cardiologists and staff members applying this procedure are exposed to high levels of scattered radiation. In our previous study the incidence of unstable chromosomal aberrations and cytokinesis-blocked micronuclei were found to be significantly higher in exposed individuals than the age and sex matched controls. In the present study we assessed cytokine production by peripheral blood mononuclear cells of the above cases and the percentage of Treg cells. According to film dosimeter analysis, personnels received 0.25-15 mSv during the previous year (average of 3 mSv/y). Isolated PBMCs from the test and control groups were stimulated with Phorbol Myristate Acetate/ Ionomycin (PMA/I). Cytokine production was measured in the supernatants of cultured lymphocytes. The percentage of Treg cells was studied by flow cytometry. The production of IL-10 and IL-5 was significantly down-regulated in the test group compared to the control group. In contrast, IL-12 was up-regulated. Yet, no statistically significant difference was found for IFN- gamma between two groups. In addition, we found higher percentage of CD4+CD25+(bright) Treg cells in the study group compared to the controls. Taken together, it was shown that low doses of scattered X-rays could skew cytokine profile of peripheral blood mononuclear cells in favour of inflammatory response causing the increase of Treg cells.
Iranian journal of allergy, asthma, and immunology 01/2008; 6(4):181-7. · 0.65 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electrotransfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.
The Korean Journal of Parasitology 01/2008; 45(4):287-93. · 0.88 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Peripheral blood mononuclear cells from subjects never exposed to Leishmania were stimulated with Leishmania guyanensis. We demonstrated that L. guyanensis-stimulated CD8(+) T cells produced interferon (IFN)- gamma and preferentially expressed the V beta 14 T cell receptor (TCR) gene family. In addition, these cells expressed cutaneous lymphocyte antigen and CCR4 surface molecules, suggesting that they could migrate to the skin. Results obtained from the lesions of patients with localized cutaneous leishmaniaisis (LCL) showed that V beta 14 TCR expression was increased in most lesions (63.5%) and that expression of only a small number of V beta gene families (V beta 1, V beta 6, V beta 9, V beta 14, and V beta 24) was increased. The presence of V beta 14 T cells in tissue confirmed the migration of these cells to the lesion site. Thus, we propose the following sequence of events during infection with L. guyanensis. After initial exposure to L. guyanensis, CD8(+) T cells preferentially expressing the V beta 14 TCR and secreting IFN- gamma develop and circulate in the periphery. During the infection, these cells migrate to the skin at the site of the parasitic infection. The role of these V beta 14 CD8(+) T cells in resistance to infection remains to be determined conclusively.
The Journal of Infectious Diseases 04/2007; 195(5):739-47. · 5.85 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Calprotectin is a calcium and zinc-binding protein complex that is abundant in the cytosol of neutrophils released under inflammatory conditions. However, the exact role of this factor has not been elucidated. It is composed of 8 and 14 kDa subunits and has the capacity to induce apoptosis in various tumor cells in a zinc-reversible manner. Reactive Oxygen Species (ROS), which are the byproducts of normal cellular oxidative process, regulates the initiation of apoptotic signaling. Recently, it has been shown that calprotectin plays an important role in phagocyte NADPH oxidase activation. In addition, the pretreatment of colon cancer cells with the antioxidant N-Acetyl-L-Cysteine (NAC) prevents apoptosis inducs by calprotectin. In the present study, we further investigate the growth inhibitory effect of calprotectin via ROS induction. For the first time it is shown that human calprotectin induced ROS and apoptosis in K562 cells revealed by conversion of Dichlorodihydroflurescin Diacetate (DCFH2-DA) to DCF and the enhancement of cell surface binding to Annexin V-FITC appropriately. More over, it is demonstrated that naturally occurring antioxidant vitamin E (50-200 μM) significantly reversed the effect of calprotectin proposing the beneficial effect of vitamin E as a natural antioxidant in restriction of calprotectin cytotoxic activity during excessive production of this protein.