Paria Heydarinezhad; Nasser Gholijani; Zahra Habibagahi; Mohammad Reza Malekmakan; Zahra Amirghofran
Abstract
Background: FOXP3, an important transcription factor of regulatory T cells has shown a contribution to the development of various autoimmune diseases. Objectives: To investigate the influence of FOXP3 polymorphisms (rs3761548 and rs2294021) on systemic lupus erythematosus (SLE) susceptibility and patients' ...
Read More
Background: FOXP3, an important transcription factor of regulatory T cells has shown a contribution to the development of various autoimmune diseases. Objectives: To investigate the influence of FOXP3 polymorphisms (rs3761548 and rs2294021) on systemic lupus erythematosus (SLE) susceptibility and patients' characteristics. Methods: Genotyping was performed on 265 patients with SLE and 404 healthy controls using PCR-RFLP. Patients' demographic, laboratory, and clinical information were all documented. The relationship between the SNPs and patients' characteristics was statistically analyzed. Results: The frequency of C/- genotype in male patients was significantly higher than in the healthy male controls, whereas the frequency of A/- genotype was lower (OR=0.53; 95% CI=0.28-1.00, p=.05). Analysis of the correlation between these SNPs and the patients' characteristics showed a longer disease duration in the rs3761548 C/- carriers and a correlation with arthralgia in both SNPs. In the females, there was a significant association between CC haplotype and disease susceptibility (OR=0.6, CI=0.38-0.94, p=.027). A significant association of both SNPs with the history of abortion was also detected. The frequencies of the rs3761548 AA (p=.006) and the rs2294021 CC genotypes (p=.038) and AC/AC combination (p=.033) were higher in women who had an abortion. We found a correlation between the rs3761548 AC genotype and the decreased C4 level and cardiovascular involvement, and the rs2294021 CC genotype with ESR, neurological involvement, and photosensitivity. Conclusions: FOXP3 rs3761548 C/- genotype association with disease susceptibility in male patients, an association of both SNPs with the abortion risk in female patients, and the correlation between these SNPs and several clinical features of the patients suggest their association with the disease development and pathology.
Fathollah Kalantar; Mohammad Hossein Dabbaghmanesh; Emanuela Martinuzzi; Mohsen Moghadami; Zahra Amirghofran
Volume 11, Issue 1 , March 2014, , Pages 1-12
Abstract
Background: Type 2 diabetes (T2D) is a chronic metabolic disorder in which beta-cells are destroyed. The islet amyloid polypeptide (IAPP) produced by beta-cells has been reported to influence beta-cell destruction. Objective: To evaluate if IAPP can act as an autoantigen and therefore, to see if CD8 ...
Read More
Background: Type 2 diabetes (T2D) is a chronic metabolic disorder in which beta-cells are destroyed. The islet amyloid polypeptide (IAPP) produced by beta-cells has been reported to influence beta-cell destruction. Objective: To evaluate if IAPP can act as an autoantigen and therefore, to see if CD8 + T-cells specific for this protein might be present in T2D patients. Methods: Peripheral blood mononuclear cells (PBMC) were obtained from human leukocyte antigen (HLA)-A2 + T2D patients and non-diabetic healthy subjects. Cells were then screened for peptide recognition using ELISPOT assay for the presence of IFN-γ producing CD8 + T-cells against two HLA Class I-restricted epitopes derived from IAPP (IAPP 5-13 and IAPP9-17) and common viral antigenic minimal epitopes Flu MP 58-66, CMV495–503, EBV280–288 and HIV77–85 as controls. Results: A total of 36.4% of patients and 56.2% of healthy subjects showed a response against IAPP 5-13 peptide. No significant difference in response against this peptide was noted between the patients and the healthy donors. With respect to peptide IAPP 9-17, although healthy subjects showed a higher mean number of spot forming cells than the patients, the difference was not significant; 36.4% of patients and 37.5% of controls responded to this peptide. The response of healthy subjects to the common viral peptides was stronger than that of the patients, though the result was not significant. Conclusions: It is unlikely that IAPP would be a target for CD8+ T-cells in diabetic patients; however, the trend observed toward a lower response of T2D patients against IAPP and common viral peptides may imply a decreased immune response in these patients.
Akram Ziaei; Zahra Amirghofran; Josef Zapp; Mohammad Ramezani
Volume 8, Issue 4 , December 2011, , Pages 226-235
Abstract
Background: Salvia mirzayanii, a native plant to Iran, is shown to have immunomodulatory effects on lymphocyte proliferation. Objective: To identify the bioactive immunomodulatory compound(s) present in S. mirzayanii. Methods: The crude extract was fractionated to five fractions in two steps using different ...
Read More
Background: Salvia mirzayanii, a native plant to Iran, is shown to have immunomodulatory effects on lymphocyte proliferation. Objective: To identify the bioactive immunomodulatory compound(s) present in S. mirzayanii. Methods: The crude extract was fractionated to five fractions in two steps using different solvents. The fractions were subjected to bioassay-guided fractionation. All the fractions were tested for bioactivity on human activated-peripheral blood lymphocytes (PBLs) using cell proliferation assay. Results: The methanol fraction (Fr. M) showed the highest inhibitory effect on PBLs compared to other fractions. Fr. M was applied on a gravity column chromatography for further fractionation. Resultant fractions, demonstrated inhibitory effects at higher concentrations. Fr. 4 with an 18.9 ± 0.2% inhibitory activity at 200 μg/ml and with the highest quantity was applied on preparative TLC plates for further purification. The final purified compound was identified as teuclatriol, a guaiane sesquiterpene, by NMR analysis. This compound showed a significant anti-proliferative effect on human activated- peripheral blood lymphocytes (IC50, 72.8 ± 5.4 μg/ml). Conclusion: Teuclatriol was found to be one of the compounds responsible for the immunoinhibitory effect of Salvia mirzayanii. We suggest further studies on teuclatriol, exploring its mechanism of action as an immunomodulatory compound.
Zahra Amirghofran; Saeed Malek-Hosseini; Hossein Golmoghaddam; Fathollah Kalantar; Mehdi Shabani
Volume 8, Issue 3 , September 2011, , Pages 159-169
Abstract
Background: A number of medicinal plants have been used to treat various immunological diseases. Nitric oxide (NO) has an important regulatory role in the various types of inflammatory processes. Objective: To investigate the NO modulatory activity of the extracts of several medicinal plants native to ...
Read More
Background: A number of medicinal plants have been used to treat various immunological diseases. Nitric oxide (NO) has an important regulatory role in the various types of inflammatory processes. Objective: To investigate the NO modulatory activity of the extracts of several medicinal plants native to Iran including Dracocephalum kotschyi, Linum persicum, Dionysia termeana, Salvia mirzayanii, Ferulago angulata and Euphorbia cheiradenia. Methods: The methanolic extracts of the plants were prepared and examined for their effects on the NO production by lipopolysaccharide-stimulated mouse macrophages. The level of TNF-α and IL-1β proinflammatory cytokines in the macrophage culture were detected using enzyme-linked immunosorbent assay. Results: All the extracts at concentration of 50 μg/ml demonstrated a significant decrease in NO production (p<0.001) after a 24-hour treatment. This inhibitory effect was also seen after 48 hours. Among the extracts, L. persicum was the strongest extract in reducing the NO production at 1 μg/ml after both 24 and 48-hours (nearly 100% inhibition, p<0.001). S. mirzayanii extract with 66.2 ± 8% inhibition at 50 μg/ml, showed the mildest effects in 48 hour culture. In cytokine release determination, the extract of L. persicum significantly inhibited both TNF-α and IL-1β cytokines production by stimulated macrophages (p<0.001). D. kotschyi, D. termeana and F. angulata decreased secretion of IL-1β from the cells. Conclusion: These results indicate the presence of anti-inflammatory and macrophage inhibitory substances in these plants.
Zahra Amirghofran; Abbas Azadmehr; Masoud Bahmani; Katayoun Javidnia
Abstract
Background: Studies have demonstrated that plant extracts possess various biological characteristics including immunomodulatory activity. Objective: Euphorbia cheiradenia Boiss et Hohen (Euphorbiaceae), a medicinal herb native to Iran was investigated for its immunomodulatory effects. Methods: The ...
Read More
Background: Studies have demonstrated that plant extracts possess various biological characteristics including immunomodulatory activity. Objective: Euphorbia cheiradenia Boiss et Hohen (Euphorbiaceae), a medicinal herb native to Iran was investigated for its immunomodulatory effects. Methods: The methanolic extract of the plant was prepared and added to mitogen-induced human peripheral blood lymphocyte cultures at different concentrations. Effect of E. cheiradenia on in vivo cell-mediated immunity was measured by delayed type hypersensitivity (DTH) reaction. The effect of the extract on humoral antibody synthesis was also measured in immunized mice treated with different extract concentrations. Results: The stimulation index (SI) for cultures treated with 0.01 to 200 microg/ml of the extract ranged from 1.3+/-0.04 to 2.4+/-0.06, (p<0.01) showing a significant stimulatory effect of E. cheiradenia on the lymphocytes. IL-2 secreted from lymphocytes treated with the extract was significantly higher than that from the non-treated cells (p<0.001). Cell cycle analysis on mitogen-treated lymphocytes exposed to different concentrations of the extract showed an increase in the percentage of cells at G2M phase with increases in the concentration of the extract, but the results was not significant. In DTH skin test, the mean footpad thickness of all mice groups treated with 1, 50 and 100 mg/kg of the extract at 24 hours after immunization with antigen was 3.5+/-0.6 mm compared to 2.5+/-0.5 mm for the non-treated group (p=0.005). Moreover, an increase in production of specific antibody in mice immunized with different extract concentrations was also demonstrated. Conclusion: Results of this study showed the ability of the E. cheiradenia extract to induce proliferation of lymphocytes and enhance both cellular and humoral specific immune responses.
Zahra Amirghofran; Abbas Azadmehr; Katayoun Javidnia
Volume 4, Issue 1 , March 2007, , Pages 26-31
Abstract
Background: Plant extracts have been widely investigated for possible immunomodu-latory properties. Objective: To study the immunomodulatory functions of the metha-nol extract of Haussknechtia elymatica (Apioideae), an herb native to south-western Iran. Methods: Delayed type hypersensitivity (DTH) skin ...
Read More
Background: Plant extracts have been widely investigated for possible immunomodu-latory properties. Objective: To study the immunomodulatory functions of the metha-nol extract of Haussknechtia elymatica (Apioideae), an herb native to south-western Iran. Methods: Delayed type hypersensitivity (DTH) skin test and measurement of an-tibody titer after immunization with Sheep-RBC was performed. [3H]-thymidine incor-poration assay on the human lymphocytes stimulated with PHA and determination of IL-2 production using ELISA method was carried out. Results: Treatment of mice with increasing concentrations of the extract decreased the footpad thickness indicating a dose-related inhibitory effect of H. elymatica on delayed hypersensitivity. The mean antibody titers for all concentrations of the extract at primary and secondary responses were significantly less than the control. Addition of the extract to the culture of human peripheral blood lymphocytes in the presence of mitogen decreased cell proliferation dose-dependently. A dose related decrease in production of IL-2 in extract-treated cells was also observed. Conclusion: The decline of antibody titer and DTH response indi-cates that H. elymatica, by acting on the lymphocyte proliferation and IL-2 secretion, inhibits both humoral and cell-mediated immune responses.
Zahra Amirghofran
Volume 2, Issue 1 , March 2005, , Pages 29-35
Abstract
Background: Perforin is known to be important in cytolytic activity mediated by natural killer (NK) cells. Objective: To study the relationship between the efficiency of NK and lymphokine-activated killer (LAK) cells activity, and the expression of perforin and HLA class I molecules. Methods: ...
Read More
Background: Perforin is known to be important in cytolytic activity mediated by natural killer (NK) cells. Objective: To study the relationship between the efficiency of NK and lymphokine-activated killer (LAK) cells activity, and the expression of perforin and HLA class I molecules. Methods: LAK cells were generated by in vitro culturing of human peripheral blood lymphocytes (PBLs) in the presence of human recombinant interleukin-2 (rIL-2). Cytotoxic activity was measured at different intervals of activation by MTT colorimetric assay using different human tumor cell lines. Immunocytochemical staining of molecules was performed on LAK/NK cells using specific monoclonal antibodies and Biotin-conjugated anti-immunoglobulin. Results: LAK/NK killing against Fen and two other cell lines, KB and Scaber showed that at day 9 and 15 of activation, 57% to 60% and 45.5% to 92.5% of Fen cells were killed at different E/T ratios. At the same time, the maximum percent killing against Scaber and KB cell lines was 47.3 and 54.3 at 5/1 ratio, respectively, showing that Fen cells were more sensitive than the two other cells. Time-course experiments using Fen cell line demonstrated 60.0, 83.9 and 34.8 percent killing at days 9, 15 and 22 at 10/1 E/T ratios. When other E/T ratios were investigated, a similar profile was observed. The maximum activity was at day 15 and 5/1 E/T ratio (92.5%). In immunocytochemical staining of activated LAK cells, 75.9% to 86.3% of LAK cells expressed HLA class I molecules. Perforin expression changed from 30.3% at day 7 to 42.7% at day 17 followed by a decrease to 27.9% at day 24. Conclusion: These data indicate that perforin expression is closely correlated with NK/LAK killing activity.
Nasser Gholijani; Ahmad Monabati; Zahra Amirghofran
Volume 1, Issue 1 , June 2004, , Pages 34-40
Abstract
Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer ...
Read More
Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer traditional prognostic indicators. Methods: In this study expression of p53 was examined in paraffin-embedded tissues from 50 cases of prostate carcinoma by immunohistochemistry and evaluated using an index of staining. Correlation between p53 expression and apoptosis was detected by TUNEL method. Pathological grade, Gleason score and stage of carcinoma were also determined. Results: P53 expression was observed in 48 of 50 cases (26-100% of tumor cells) with mean staining index of 141±65. A significant association between p53 expression and pathologic grade (r=0.37, p=0.004) and Gleason score (r= 0.4, p=0.009) of patients was observed. Apoptosis was detected in only 6 patients. p53 expression showed no correlation with apoptotic index. No correlation between p53 expression and stage or apoptosis and clinicopathological characteristics of patients was found. Conclusion: p53 expression showed a significant correlation with differentiation status of the prostate carcinoma and can be helpful as a prognostic marker. Decreased level of apoptosis observed in our cases was not correlated with p53 expression indicating the possible role of other regulatory molecules involved in the apoptosis.
