فهرست مطالب سیدمحمدموذنی

Interaction of cytomegalovirus glycoprotein B with toll-like receptors of dendritic cells leads to early signaling and innate immune responses. The aim of this study is to evaluate the effects of cytomegalovirus glycoprotein B on the maturation and function of monocyte-derived dendritic cells in treated groups in comparison with control groups. Blood samples were taken from 5 healthy volunteers. Following the generation of monocyte-derived dendritic cells on the fifth day of cell culture, half of the immature dendritic cells were treated with cytomegalovirus glycoprotein B, and the rest of them were induced to mature dendritic untreated cells and were used as the control group. The maturation and function of dendritic cells were evaluated in these two groups. The gene expression level of toll-like receptor-4 significantly increased in the group treated with glycoprotein B (p < 0.05), whereas there were no significant differences in the expression rates of CD83, CD86, CD1a, and HLA-DR and the secretion of IL-23 from monocyte-derived dendritic cells between the treated groups and the controls. The increase in the gene expression of toll-like receptor-4 in monocyte-derived dendritic cells treated with cytomegalovirus glycoprotein B showed that cell contact is required to elicit cellular antiviral response and toll-like receptor activation. Thus, it is critical to recognize the viral and cellular determinants of the immune system in order to develop new therapeutic strategies against cytomegalovirus.

Macrophages play a crucial role in appropriate embryo implantation and maintenance of pregnancy. This study was done to evaluate the frequency and distribution of uterine (local changes) and spleen (systemic changes) macrophages on days 4.5 and 7 of NMRI mouse pregnancy which are simultaneous with beginning of implantation and complete establishment of embryo in the uterus. The precise day of pregnancy was determined through vaginal plug detection. Blood was collected from pregnant mice on days 4.5 and 7 of gestation and serum 17- β estradiol and progesterone concentrations were measured using the ELISA method. The frequency and localization of macrophages in different regions of uterus and spleen were also investigated by immunohistochemistry staining using anti F4/80 antibody. The results of this study showed that frequency of uterine macrophages has decreased on day 7 in comparison to day 4.5 of gestation. Macrophages are distributed throughout the whole uterus on day 4.5 while their distribution was restricted to myometrium and stroma beneath the circular muscles on day 7 of pregnancy. The frequency of spleen macrophages as a representative of their systemic changes was not different in studied groups. Serum 17-β estradiol and progesterone concentrations increased significantly on day 7 compared to day 4.5 of pregnancy. Considering the changes in 17-β estradiol and progesterone concentrations and their indirect effects on macrophages recruitment through regulation of M-CSF and GM-CSF secretion by uterine stromal cells, the differences in frequency and distribution of macrophages in different stages of pregnancy could be explained. While the macrophages chemotactic factors and homing receptors and their sources in spleen are different from uterus and don’t response to ovarian hormones variations. Regarding the role of macrophages in embryo implantation and regulation of feto-maternal immune responses, it seems that the changes in their frequency and localization can be helpful for a successful pregnancy.

One of the common causes of recurrent spontaneous abortion is interference of immunological factors. Administration of cyclosporine, as an immunosuppressive drug, has reportedly reduc the rate of abortion in abortion-prone model of mice CBA/J mated DBA/2 male mice. On the other hand, asymmetric antibodies and TGF-β1 have been considered as an important factor in successful pregnancy outcomes. Intravaginal administration of TGF-β has also been reported to reduce the resorption rate in the same mouse model. In this study, the immunomodulatory effects of cyclosporine on serum level of asymmetric antibodies (AAbs) and T cell growth factor beta (TGF-β1) were investigated. Female CBA/J mice were mated with male DBA/2. The pregnant females were divided into two groups: 1- test group; received intraperitoneal injection of 1mg/kg cyclosporine (n=7), 2- control group; received intraperitoneal injection of Phosphate Buffered Saline (PBS) (n=7). Both groups received treatments on the 5th day of pregnancy. Serum samples were collected on the 13.5th day of pregnancy, when the percentage of abortion was also calculated. The percentage of AAbs and abortion rate were significantly reduced in cyclosporine group (P<0.05, for both variables), while the serum levels of TGF-β1 did not show significant difference between groups (P>0.05). Our study showed that despite the significant reduction in percentage of AAbs and no significant difference in serum level of TGF-β1, the abortion rate was markedly reduced in cyclosporine group. So other mechanisms of immunomodulation and reduction of abortion rate except asymmetric antibodies and TGF-β1 should be involved in this mouse model following cyclosporine administration.

The anti-inflammatory effects of the vitamin D and ginger have been demonstrated in some studies. The aim of this study was to evaluate the effects of vitamin D and ginger extract on the clinical symptoms and leukocyte infiltration of experimental autoimmune encephalomyelitis (EAE). For the induction of EAE, the (C57BL/6) were immunized subcutaneously MOG peptide emulsified in complete Freund's adjuvant. The mice divided into EAE group includimg: vitamin D-treated, ginger extract-treated, olive oil and phosphate-buffered saline, and also one group which was considered as normal group and received only PBS(phosphat- Buffered Saline). The mice were sacrificed at day 31 after immunization and the leukocytes infiltration was evaluated in the CNS (central nervous system). The EAE scores and the body weight were evaluated till day 30. Data were analyzed using t-test. Two control groups showed the clinical symptoms at day 10, whereas vitamin D and ginger-treated groups exhibited the symptoms at days 13 and 15, respectively. The maximum mean score of disease was (1.87±1.61), (1.92±1.79), (1.05±1.01), and (0.73±0.81) for olive oil-, PBS-, vitamin D3-, and ginger-treated groups, respectively. Vitamin D3 and ginger extract increased the body weight significantly)P =0.05) as compared to control groups. The leukocytes infiltration was also significantly lower in treated groups as compared to control group. These observations represent that the vitamin D3 and ginger extract have significant capability to attenuate the EAE severity.

The important role of co-stimulatory molecules in immunological responses has been clearly demonstrated. With the development of new methods in molecular immunology, preventing the co-stimulatory signals in order to induce tolerance has emerged as a novel strategy. RNA interference is a new method that can specifically and effectively reduce target gene expression. In this study, we used a lentiviral system to target the expression of CD40 gene in dendritic cells. ShRNA sequence is able to reduce the expression of CD40 gene on murine dendritic cells. Our results indicate 64-fold decreased expression of CD40 mRNA in the group receiving lentiviral shRNACD40 as well as 31% decrease in expression of CD40 protein on the surface of dentridic cells (p<0.0001). Also, dendritic cells cannot stimulate T cells upon contact with them. This study indicates that a decrease in expression of CD40 molecule induces tolerogenic dendritic cells and prevents Th1 immunological response. Prevention of Th1 response would shift the immunological responses to Th2 response, and this type of response can be effective in autoimmune diseases including rheumatoid arthritis and type I diabetes. References 1- Groux H, Fournier N, Cottrez F. Role of dendritic cells in the generation of regulatory T cells. Sem in Immunol. 2004; 16: 99-106. 2- Kanako L, Reizis L, Reizis B. Dendritic Cells: Arbiters of Immunity and Immunological Tolerance. Cold Spring Harb Perspect Biol. 2012; 4: a007401. Avalable from: www.cshperspectives.org. 3- Morel PA. Dendritic cell subsets in type1diabetes: friend or foe? Frontiers in Immunol. 2013; 4. Avalable from: http://dx.doi.org/ 10.1155/ 2013/972865. 4- Machen J, Harnaha JO, Lakomy R, Styche A, Trucco M, Giannoukakis N. Antisense oligonucleotides down-regulating costimulation confer diabetes-preventive properties to nonobese diabetic mouse dendritic cells. The J Immunol. 2004; 173: 4331-41. 5- Maldonado R, Andrian UH. How tolerogenic dendritic cells induce regulatory T cells. Adv Immunol. 2010; 108: 111-165. 6- Peters AL, Stunz LL, Bishop GA. CD40 and autoimmunity: the dark side of a great activator. Sem Immunol. 2009; 21: 293-300. 7- Li L, Wang H, Wang B. Anergic cells generated by blocking CD28 and CD40 costimulatory pathways in vitro ameliorate collagen induced arthritis. Cell Immunol. 2008; 254: 39-45. 8- Zheng X, Suzuki M, Zhang Z, et al. RNAi-mediated CD40-CD154 interruption promotes tolerance in autoimmune arthritis. Arth Res & Ther. 2010; 12: R13. 9- Pletinckx K, Dohler A, Pavlovic V, Lutz M.B. Role of dendritic cells maturity/ costimulation for generation, homeaostasis and suppressive activity of regulatory T cells. Fronties in Immunol. 2011; 2: 7-22. 10- Gavrilov K, Saltzman WM. Therapeutic siRNA: Principles, challenges and strategies. Yale J Biol Med. 2012; 85(2): 187-200. 11- Manjunath N, Wu H, Subramanya S, Shankar P. Lentiviral delivery of short hairpin RNAs. Adv Drug Delivery Rev. 2009; 61: 732-45. 12- Inaba K, Inaba M, Romani M, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with GM-CSF. J Exp Med. 1992; 176: 1693-702. 13- Virus packaging protocol. Stem Cell Biotechnology Research Center protocol, Iran. 2013. Avalable from: www.stemcellstech. com/index.php/fa/. 14- Muller G, Muller A, Jonuleit H. Fetal calf serum –free generation of functionally active murine dendritic cells suitable for invivo therapeutic approaches. J Inv Dermatol. 2000; 114: 142-8. 15- Marin-Gallen S, Clemente-Casares X, Planas R, et al. Dendritic cells pulsed with antigen-specific apoptotic bodies prevent experimental type 1 diabetes. Clin and Exp Immunol. 2009; 160: 207-14. 16- Hasse C, Ejrnaes M, Juedes AE, Wolf T. Immunomodulatory dendritic cells require autologous serum to circumvent nonspecific immunosuuressive activity in vivo. Blood. 2005; 106: 4225-33. 17- Feili-Hariri M, Dong X, Alber SM, Watkins SC, Salter RD, Morel PA. Immunotherapy of NOD mice with bone marrow–derived dendritic cells. Diabetes. 1999; 48: 2300-08. 18- Karimi MH, Ebadi P, Pourfathollah AA, et al. Immune modulation through RNA interference-mediated silencing of CD40 in dendritic cells. Cellular Immunol. 2009; 259: 74-81. 19- Mei S, Jin-Dong LI, Rui J, et al. Construction and characterization of lentiviral shRNA expression vector targeting rat CD40 gene in dendritic cells. Chem Res. 2009; 25(5): 666-72. 20- Jantsch J, Turza N, Volke M, et al. Small interfering RNA (siRNA) delivery into murine bone marrow-derived dendritic cells by electroporation. J of Immunological Methods. 2008; 337: 71-7. 21- Giannoukakis N, Trucco M. Dendritic cell therapy for type 1 diabetes suppression. Immunother. 2012; 4(10): 1-12. 22- Giannoukakis N. Tolerogenic dendritic cells for type 1 diabetes. Immunother. 2013; 5(6): 1-3. 23- Lee CN, Lew AM, Wu L. The potential role of dendritic cells in the therapy of Type1 diabetes. Immunother. 2013; 5(6): 591-606.

Recent evidences suggest that tumors arise from a small subpopulation of cells، the cancer stem cells (CSCs) or tumor initiating cells. CSCs are able to resist the conventional methods of cancer therapy due to existence of ABC transporters on their surface. This leads to CSC resistance and maintenance resulting in post-treatment relapse and metastasis. Therefore، precise identification and characterization of these cells as a target for new therapeutic regimens is the goal of numerous studies. This study، with the intent to design a new method of immunotherapy for targeting cancer stem cells in mouse malignant melanoma، initially characterized the cancer stem cells in this malignancy. In order to identify the CSCs we induced a melanoma tumor using the B16F10 cell line in C57BL/6 mice. The tumor bulk was dissociated by an enzymatic method and homogeneous tumor cells were sorted using anti-CD44 and anti-CD24 antibodies. The sorted tumor cell subpopulations were compared according to their ability to form cell spheres in serum free medium (SFM). We determined the tumor formation ability of all cell subpopulations by transplanting serial dilutions of B16-F10 and all sorted cells sub-populations into C57/BL6 mice. The results showed that although all separated cell subpopulations and B16-F10 cells formed non-adherent spheroids in SFM in the presence of B-27، but the CD24+ cells presents a significantly higher ability to produce spheroids. The B16F10 cell line، CD44+CD24- and CD44-CD24- cells showed equal potencies in tumor induction (1 in 21730 cells). The CD44-CD24+ cells tumor induction potency was 1 in 17426 and this ability for the double positive cells (CD44+CD24+) was 1 in 11295. Collectively،the double positive (CD24+CD44+) cells were more potent in both spheroid formation and tumorogenicity. Hence they might be the CSC population of mouse melanoma.

Abstract The present study aimed to investigate the immunomodulatory activity of molecules secreted by decidual cells on dendritic cells (DCs) function in abortion-prone compared with non-abortion-prone mice. The decidual cell supernatants (DS) were obtained from abortion and non-abortion mouse models. DCs were purified from CBA/J mice spleens and treated with antigen and DS. Treated DCs were injected into mice palms. After 5 days، draining lymph nodes were removed، cultured in the presence of cognate antigen، and proliferation of lymphocyte cells was measured by 3H-thymidin incorporation. Our results showed that immunosuppressive activity of DS from non-abortion-prone mice significantly decrease dendritic cells'' ability to stimulate lymphocytes proliferation compared with DS from abortion-prone mice (Simulation index (SI) of 4. 93 ± 0. 34 versus 11. 84 ± 0. 79). We، also found that DS prepared from non-resorption sites compared with DS from resorption sites in abortion-prone mice had increased immunosuppressive activity on DC function (SI of 7. 31 ± 1. 02 versus 2. 67 ± 0. 49). Due to our results، we concluded that immunomodulatory activity of molecules secreted within decidual tissue is different between abortion-prone and non-abortion-prone mice. Based on the key role of DCs in inducing fetomaternal tolerance، we claimed that these molecules، through modulation of DCs function، play crucial role on pregnancy outcome.

As the main virulence factor of Helicobacter pylori, HP-NAP has an important role in immunoprotection against this pathogen. This antigen is a strong candidate as a part of multi-component vaccine in the clinical trial against this bacterium. Due to NAP importance, it was used in this study as a template for optimization of heterologous genes with a low A-T content and low expression in E. coli. A synthetic single gene that could reach the highest level of expression in the host was designed by using bioinformatics tools. A number of factors that influence gene expression level were changed for HP-NAP gene optimizing: the codon usage bias in E. coli was changed; the G+C content was upgraded from 38% to 45%; and the stem-loop structure was broken. These could result to prolong of the half-life of the mRNA and overexpression of recombinant of HP-NAP protein up to 800 mg per liter. Applying of bioinformatics tools was appropriated to optimize of HP-NAP overexpression in E. coli. From our results, it appears that combination of In Silico and experimental approach is a logical approach for expression of heterologous genes in another host.

Common variable immunodeficiency (CVID) is one of the most frequent cases of primary immunodeficiency. It is likely that this heterogeneous disease is caused by several distinct genetic disorders. The activation-induced cytidine deaminase (AID) enzyme is involved in class switching, somatic hypermutation (SHM) and processes associated with gene conversion in the germinal center. In order to clarify the possible role of AID in the pathogenesis of CVID, we have studied the AID gene expression in CVID patients. Peripheral blood mononuclear cells (PBMC) from 21 patients and healthy controls were isolated. The isolated cells were stimulated by CD40L and IL-4 to induce AID gene expression. After five days, total RNA from the stimulated cells was extracted and AID gene expression was investigated by RT-PCR. RT-PCR results showed that after stimulation by CD-40L and IL-4, the AID gene was expressed in all of the samples. The control samples were also positive for AID gene mRNA expression. In this investigation we studied the expression of AID gene in CVID patient's B lymphocytes for the first time. Regards to our results which showed that all patients normally expressed the AID gene mRNA and considering that one of the main problems in a number of CVID patients is disorders in phenomena related to the germinal center and complete differentiation of B lymphocytes, it can be concluded that possible defects in other molecules involved in class switching is responsible for this disease. Understanding the various genetic defects responsible for this heterogeneous disease could lead to its division into more homogenous subtypes with distinct therapeutic strategies, so further investigations is recommended.

Cancer as the uncontrolled proliferation and spread of transformed cells is one of the leading causes of mortality in the world. Dendritic cells (DCs) can be used in cancer immunotherapy. Tumor antigen-loaded DCs are able to enhance the cytotoxic T cells (CTLs) antitumor activity. Related articles were searched by search engines in NCBI database. Then the original and review articles were retrieved from the Science Direct، Nature، Wiley-Blackwell، Springer and ProQuest databases. The DC subsets with various functions have been identified in different tissues of both the animal model (mouse) and human. Human myeloid DC subsets could be derived from blood monocytes using the cytokine enriched media. Mouse conventional CD8+ DCs can be isolated from the spleen or derived from the bone marrow stem cells using the cytokine- enriched media. Some of the human and mouse DC subsets، including human myeloid DCs and mouse conventional CD8+ DCs play a pivotal role in an induction of TH1 cells and CTLs. Therefore، these subsets can be utilized as cellular immunity stimulants in human and mouse model for cancer immunotherapy.

Enterotoxigenic Escherichia coli is the most common bacterial agent producing diarrhea in the world wide. This bacterium includes some virulence factors such as; colonization factors and heat-labile toxin and/ or heat-stable toxin. Enterotoxigenic Escherichia coli attached to the intestinal epithelial cell surface through CFs and causes diarrhea by toxin secretion. Molecular pathogenesis stu-dies demonstrate that there are many pro-phylactic approaches for this infection. The best possible way is designing and prod-ucing efficient vaccine that enabeling to induc protective immunity against the most of bacterial strains. Vaccine candidate molecule (s) that are used to design a vaccine، in addition to possess immuno-genicity and safety; they should be wid-espread among variety of strains and co-ntain conserved and common epitops. This article pays attention to review some infor-mation and results of conducted researches on enterotoxigenic Escherichia coli vaccine and importance of the bacteria and its pathogenesis.