Hadi Reihani; Maryam Rastin; Mahmoud Mahmoudi; Mohsen Ghoryani; Nafiseh Abdollahi; Nafiseh Sadat Tabasi; Shahrzad Zamani Taghizadeh Rabe; Maryam Sahebari
Volume 12, Issue 2 , June 2015, , Pages 82-93
Abstract
Background: Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease. Emerging data suggests that T helper 17 (Th17) cells play a pathogenic role in SLE and the increased number of these cells correlates with disease activity. In recent years, 1α, 25-dihydroxyvitamin D3 (1,25VitD3) ...
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Background: Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease. Emerging data suggests that T helper 17 (Th17) cells play a pathogenic role in SLE and the increased number of these cells correlates with disease activity. In recent years, 1α, 25-dihydroxyvitamin D3 (1,25VitD3) has been considered as an immunomodulatory factor. Objective: To investigate the effect of 1,25VitD3 on Th17 cells and on the expression of related cytokines in SLE patients. Method: Thirty SLE patients (newly diagnosed or in remission) were sampled for 10 ml whole blood to isolate peripheral blood mononuclear cells (PBMCs) using Ficoll-Hypaque density gradient centrifugation. Isolated cells were cultured in the presence and absence of 50 nM 1,25VitD3. After incubation, cells were harvested and stimulated for 4-5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. IL-17 secreting cells were analyzed by flowcytometry. RNA was extracted from cultured cells, cDNA was synthesized, and the expression levels of IL-6, IL-17, IL-23 and TGF- β genes were assessed by real-time PCR. Results: The percentage of Th17 cells (CD3+ CD8- IL-17+ T cells) decreased significantly in 1,25VitD3-treated cells (3.67 ± 2.43%) compared to untreated cells (4.65 ± 2.75%) ( p=0.003). The expression of TGF- β up regulated (1.38-fold) and the expression of IL-6 (50%), IL-17 (27%) and IL-23 (64%) down regulated after 1,25VitD3 treatment. Conclusion: This study showed that 1,25VitD3 modulates Th17 related pathways in SLE patients and revealed the immunomodulatory effect of 1,25VitD3 on the Th17 mediated autoimmunity.
Fatemeh Vahedi; Mahmoud Reza Jaafari; Mahmoud Mahmoudi
Volume 7, Issue 4 , December 2010, , Pages 210-216
Abstract
Background: DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Objective: Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen (PA) of ...
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Background: DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Objective: Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen (PA) of Bacillus anthracis (B. anthracis) into cationic liposomes, and evaluation of its effect on specific humoral specific immunity against PA were aimed. Methods: The liposomes containing pcDNA3.1+PA plasmids were prepared with phosphatidylcholine (PC), dioleoyl phosphatidylethanolamine (DOPE) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) using dehydration-rehydration method. BALB/c mice were immunized by intramuscular (IM) injection to investigate the immunogenicity of the formulations. The resulting specific antibodies against PA, total IgG, IgG1, IgG2a and IgG2b isotypes, were evaluated by enzyme linked immunosorbent assay (ELISA) method. Conclusion: A higher concentration of specific IgG against PA was found in sera of a group immunized with the encapsulated plasmid compared with the naked plasmid alone. This difference was significant for IgG1 isotype.
Mohammad Fereidouni; Farhzad Jabbari Azad; Mahmoud Mahmoudi; Abdolreza Varasteh; Reza Farid Hosseini
Volume 7, Issue 1 , March 2010, , Pages 1-7
Abstract
Background: Invariant natural killer cells (iNKT) are an important immunoregulatory T cell subset. Currently several flow cytometry-based approaches exist for the identifi-cation of iNKT cells, which rely on using the 6B11 monoclonal antibody or a combina-tion of anti-Vα24 and anti-Vβ11 antibodies. ...
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Background: Invariant natural killer cells (iNKT) are an important immunoregulatory T cell subset. Currently several flow cytometry-based approaches exist for the identifi-cation of iNKT cells, which rely on using the 6B11 monoclonal antibody or a combina-tion of anti-Vα24 and anti-Vβ11 antibodies. Objective: The aim of this study was to compare the ability of two flow cytometry-based methods for detecting the frequency of circulating iNKT cells. Methods: The frequency of iNKT cells was detected in the pe-ripheral blood of 37 healthy adult donors by flow cytometry using the 6B11 antibody or a combination of anti-Vα24 and anti-Vβ11 antibodies. Results: The frequency of iNKT cells detected by 6B11 antibody or by combination of anti-Vα24 and anti-Vβ11 anti-bodies was significantly different (0.54% vs. 0.31%, respectively, p<0.001) but the val-ues were highly correlated (Spearman r = 0.742, p<0.0001). Conclusion: The results of this study indicate that different combinations of mAbs detect different frequencies of peripheral blood iNKT cells and a consensus in the field needs to be established to al-low better assessment of iNKT-related studies and suggest using different methods for accurate identification of iNKT cells.
Zahra Meshkat; Hoorieh Soleimajjahi; Mahmoud Mahmoudi; Zuhair Mohammad Hassan; Hessam Mirshahabi; Mojtaba Meshkat; Maryam Kheirandish
Abstract
Background: Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among female population worldwide. Specific human papillomaviruses and, most notably, HPV types 16 and 18 are recognized as being caus-ally associated with cervical carcinomas. The early ...
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Background: Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among female population worldwide. Specific human papillomaviruses and, most notably, HPV types 16 and 18 are recognized as being caus-ally associated with cervical carcinomas. The early HPV type 16 genes, E6 and E7, di-rectly participate in the in vitro transformation of primary human keratinocytes and rep-resent an excellent target for immune therapy of HPV related disease. Objective: The aim of this study was the evaluation of the efficacy of a DNA vaccine containing human papillomaviruse type 16 E7 gene (Iranian isolate) in induction of CTL responses in an animal model. Methods: In this study, the expression vector containing HPV type 16 E7 gene was constructed and chosen as a model antigen in the development of a thera-peutic DNA vaccine in an animal model. CTL responses, cytokine assay, lymphocyte stimulation test, CD4 and CD8 staining and flowcytometry were done for evaluating of the immune responses. Results: Our findings indicate that the target DNA vaccine can induce an E7-specific CTL response, which is important in the lysis of infected tumor cells, compared to negative control (p<0.005) after in vivo immunization in the mouse system. Conclusion: The developed vaccine may be promising as an anti-cancer vac-cine.
