Samira Rajaei; Amir Hassan Zamani; Mahmood Jeddi-Tehrani; Maryam Tavakoli; Afsaneh Mohammadzadeh; Ali Dabbagh; Mahroo Mirahmadian
Volume 8, Issue 4 , December 2011, , Pages 201-208
Abstract
Background: Repeated Implantation Failure (RIF) is one of the most intricate obstacles in assisted reproduction. The cytokine and chemokine composition of uterine cavity seem to play important roles in the implantation process. Objective: To compare the cytokine profile in the endometrium of normal fertile ...
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Background: Repeated Implantation Failure (RIF) is one of the most intricate obstacles in assisted reproduction. The cytokine and chemokine composition of uterine cavity seem to play important roles in the implantation process. Objective: To compare the cytokine profile in the endometrium of normal fertile women and those with repeated implantation failure. Methods: After enzymatic digestion of endometrial tissues, whole endometrial cells and endometrial stromal cells from RIF and normal fertile women were cultivated and stimulated for cytokine secretion. The levels of IL-10, TGF-β, IFN-γ, IL-6, IL-8 and IL-17 in culture supernatants of the two groups were assayed by ELISA and compared together. Results: Endometrial stromal cells and whole endometrial cells of normal fertile women produced higher levels of IL-6, IL-8 and TGF-β compared to RIF group, although this difference was statistically significant only in endometrial stromal cells (p=0.005, 0.002 and 0.001, respectively). In addition, endometrial stromal cells of normal fertile women produced lower levels of IL-10 in comparison with RIF group (p=0.005). Conclusion: Disturbances in cytokine production at the feto-maternal interface could be a cause of implantation failure. A pro-inflammatory cytokine milieu seems to be pivotal for successful implantation.
Mohammad Jafar Mahmoudi; Maryam Mahmoudi; Fereydoon Siassi; Fazel Shokri; Mohammad Reza Eshraghian; Amir Hassan Zamani; Reza Chahardoli; Mona Hedayat; Jalal Khoshnoodi; Hashem Nayeri; Nima Rezaei; Ali-Akbar Saboor-Yaraghi
Volume 8, Issue 1 , March 2011, , Pages 27-33
Abstract
Background: Atherosclerosis, a chronic inflammatory disease of the vessel wall is characterized by local and systemic immune responses to a variety of antigens. Oxidized low-density lipoprotein (oxLDL) is considered as an important determining factor in the pathogenesis of atherosclerosis. Objective: ...
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Background: Atherosclerosis, a chronic inflammatory disease of the vessel wall is characterized by local and systemic immune responses to a variety of antigens. Oxidized low-density lipoprotein (oxLDL) is considered as an important determining factor in the pathogenesis of atherosclerosis. Objective: The purpose of this study was to investigate the degree of peripheral blood mononuclear cells (PBMC) vulnerability to in vitro oxLDL-induced cytotoxicity from atherosclerotic patients in comparison to healthy individuals. Methods: Thirty patients with atherosclerotic lesions, confirmed by angiography, and 30 matched healthy individuals were investigated. PBMC was prepared from individuals' blood samples which were further stimulated with low dose (1 μg/mL) and high dose (50 μg/mL) of extensively oxidized LDL. MTT assay was utilized to measure cell viability and proliferation. Stimulation index (SI) was calculated as mean ratio of optical density (OD) of the stimulated cells divided by OD of untreated cells. Results: Low dose oxLDL treatment caused no significant proliferative or cytotoxic effect in the control group; however, similar treatment caused significant cytotoxic effect in the patient group compared to the controls (p=0.026). High dose oxLDL treatment induced more significant cytotoxicity in the patient compared to the control group (p=0.006). Comparison of the SI between the two groups of patients and controls showed significantly lower index by either the low (p=0.03) or the high dose (p<0.001) oxLDL in the patients compared to the controls. Conclusions: PBMC from patients with atherosclerosis showed increased susceptibility to oxLDL-induced cytotoxicity. Our results imply that prolonged exposure to elevated levels of circulating oxLDL could weaken the cellular defense mechanisms by progressive depletion of the pool of antiapoptotic proteins, rendering the cells more vulnerable to oxLDL-induced cell death.
Mohammad Mahdi Eftekharian; Amir Hassan Zamani; Seyed-Mohammad Moazzeni
Volume 7, Issue 2 , June 2010, , Pages 74-82
Abstract
Background: Dendritic cells (DCs) play a central role in the initiation and expansion of T cell mediated immune responses with potential immunotherapy application. The compounds which have the ability to induce immunomodulatory effects on DCs may be employed for the treatment of immunopathologic conditions ...
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Background: Dendritic cells (DCs) play a central role in the initiation and expansion of T cell mediated immune responses with potential immunotherapy application. The compounds which have the ability to induce immunomodulatory effects on DCs may be employed for the treatment of immunopathologic conditions such as autoimmune diseases. Objective: The aim of this study was to investigate the in vivo effects of calcitriol (active form of vitamin D3) on DCs. Methods: 0.1 microgram calcitriol was injected intra-peritoneally into C57BL/6 mice every other day within 3 weeks, and spleen DCs were extracted by magnetic beads. The phenotypic and functional properties of DCs were studied by flow cytometry and mixed lymphocyte reaction (MLR), respectively. Results: The expression of CD86 and MHC II, as maturation markers and costimulatory molecules were significantly decreased (p=0.028 and p=0.047, respectively) while CD11b expression, as a marker of mice myeloid DCs which mostly induces Th2 cytokine profile, was significantly increased (p=0.011). Allogeneic T cell stimulation in MLR was also significantly inhibited in comparison with the control groups (p<0.05). Conclusion: Our data indicate that in vivo calcitriol administration inhibits maturation and activation of DCs in the same manner as in vitro conditions.
Sorour Shojaeian; Amir Hassan Zamani; Mahmood Jeddi-Tehrani; Forouzandeh Fereidooni; Ebrahim Torkabadi; Mohammad Mehdi Akhondi; Akram Sadat Tabatabaei-Panah; Abdolamir Allameh
Volume 6, Issue 4 , December 2009, , Pages 174-185
Abstract
Background: Monoclonal antibodies (mAbs) are essential tools for many molecular im-munology investigations, epitope mapping and molecular modelling, clinical laboratory di-agnostic tests and immunotherapy. Humoral immune response of immunized animals largely depends on the nature of antigen and the immunization ...
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Background: Monoclonal antibodies (mAbs) are essential tools for many molecular im-munology investigations, epitope mapping and molecular modelling, clinical laboratory di-agnostic tests and immunotherapy. Humoral immune response of immunized animals largely depends on the nature of antigen and the immunization technique. Polysaccharides and heavily-glycosylated proteins are very elusive targets incapable of mounting long-lasting, high affinity antibody responses. Carcinoma antigen 125 (CA 125), a well known tumor marker of ovarian cancer, is a mucin type antigen consisting of repetitive units of heavily glycosylated moieties which render production of mAbs very difficult. Objective: To evaluate the efficacy of heterologous antigen preparations as a way of mouse immuniza-tion in the production of anti-CA 125 mAb. Methods: Two different protocols of immuni-zation were used for priming of NMRI mice. In the first method, mice conventionally im-munized by three intraperitoneal injections of purified CA 125 and boosted by the antigen three days before fusion. In the second approach, mice were primed by three intraperitoneal injections of living CA 125 positive cells of OVCAR-3 cell line, and boosted by intrave-nous injection of the purified extracellular domain of CA 125. Production of mAb was per-formed by standard hybridoma technology and mAbs were characterized by different im-munoassays. Results: The first method failed to produce stable clones despite six time fu-sion. A total of ten stable clones, however, were produced in the second approach. Some of the clones were characterized and found to have excellent immunoreactivity when tested by ELISA assay, western blotting, intracellular and surface immunofluorescent staining of OVCAR-3 cell line and immunohistochemical staining of ovarian cancer tissues. Conclusion: Altogether the results of the present study clearly showed that heterologous antigen preparation is the method of choice for immunization when production of mono-clonal antibody against highly glycosylated poorly immunogenic antigens is concerned.
