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Background: Common variable immune deficiency (CVID) is the commonest symptomatic primary immunodeficiency and represents a heterogenous collection of disorders resulting mostly in antibody deficiency and recurrent infections .The syndrome includes impaired B-cell maturation, impaired somatic hypermutation, reduced numbers of circulating memory and isotype-switched memory B cells, and absent or reduced plasma cells. B cell maturation antigen (BCMA) is a tumor necrosis family receptor superfamily member 17 (TNFRSF17), expressed only on B cell lines , and is essential for survival of long-lived plasma cells .The aim of this study was to evaluate mutations in BCMA in patients with CVID in compare with normal individuals in Isfahan, Iran. Methods: Blood samples were collected from 10 CVID patients with substitutive immunoglobulin therapy before immunoglobulins (Ig) infusion and 10 normal controls in ethylenediaminetetraacetic acid (EDTA) tubes then DNA samples were extracted and after the polymerase chain reaction (PCR) was done, samples were sequenced. Findings: After reviewing the results of the sequence and alignment of the sequences, no mutations in the gene were seen. Conclusion: In addition to the study of mutation in BCMA gene, BCMA gene and protein expression level should be considered to understand more aspects of this disease.

Background: Common variable immunodeficiency (CVID) is a heterogeneous set of immunological abnormalities including decreased serum levels of antibodies (hypogammaglobolinemia), at least for two isotopes of immunoglobulin, and impaired antibody response to infection or vaccination. Thus, it is associated with increased susceptibility to infections. This study aimed to investigate the frequency of B cells in the patients with CVID in Isfahan city, Iran. Methods: Blood samples were collected from the patients with substitutive immunoglobulin (Ig) therapy before immunoglobulin infusion. Then, peripheral blood mononuclear cells (PBMC) were isolated via Ficoll-Hypaque density centrifugation. Flow cytometry method was employed to determine the frequency and phenotype of the B and transitional B cells using the antibodies of CD19-FITC (Exebio), CD24-PE (Exebio), and CD38 PE (BD Exebio). Findings: There was significant difference in the proportion of the peripheral blood B cells in the patients with CVID, compared to the healthy subjects (P < 0.001). But, there was insignificant difference in the proportion of the transitional B cells in the patients, compared to the healthy subjects (P = 0.267). Conclusion: The results show that there is insignificant difference in the proportion of the transitional B cells in the patients with CVID, compared to control group. However, more studies should be carried out concerning mutation in the involved genes to understand more aspects of this disease.

Objective(s): siRNA may be a very promising tool for treatment of various diseases especially in cancer therapy due to high specificity. One of the main hurdles applications of siRNAs in vivo is optimization of the delivery strategy, especially the carrier systems. The aim of this study was to optimize siRNA delivery into suspended erythroleukemic cell line K562. Materials and Methods: We applied polyethyleneimine (PEI) and oligoethyleneimine (OEI) derivatives alone or their co-formulation with different agents such as chloroquine (a drug known to alter lysosomal pH and thus to inhibit lysosomal degradation of macromolecules), DOPE (lipophilic agent), succinic acid (introduction of negatively charged to polymer) and transferrin (the ligand of transferring receptor which is over-expressed in many types of tumors and hematopoietic cells). Results: In this study it was shown that utilizing a combination of 70% OEI-HA10 (ten hexyl acrylate residues per one OEI chain) plus 30% of transferin-PEI with Luc-siRNA was highly effective for transfecting K562 cell. This co-formulation silenced luciferase activity up to 70% after short time without any significant inhibition in the luciferase activity in siCONTROL wells. Conclusion: In conclusion, the combination of modified PEI with transferrin and OEI by hexyl acrylate may increase siRNA delivery and reduce toxicity in hematopoietic suspended cells.

Background: Transformation and differentiation of activated B-cell to plasmacells and also memory cells depend on signaling from B-cell receptors. The signals from antigen and cytokine receptors on the surface of B cells lead to induce the expression of specific transcription factors, which finally determine the fate of B cells. Methods: Peripheral blood mononuclear cells (PBMC) were isolated via ficoll gradient and then purified B cells were separated using magnetic-activated cell sorting (MACS). Pure B cells were cultured in Roswell Park Memorial Institute 1640 (RPMI1640) culture media at the presence of purified anti-human CD40 antibody and anti-immunoglobulin M f (ab)´2 or lipopolysaccharide (LPS) and anti-human CD40 antibody that induced B-cells differentiation to plasmablasts which was assessed with 3 markers (CD38+, CD27+, IgM-) and analyzed via flow cytometry. Findings: In stimulation of B cells with purified anti-human CD40 antibody and anti-IgM f (ab)´2 or LPS through cross-linking B-cell receptor, the majority of B cells remained alive and differentiated to another lineage of B cells (plasmablast: CD38+, CD27+, IgM-). There was no significant statistical difference between expressions of plasmablast markers in two states of stimulation. Conclusion: B cells can be stimulated and differentiated to plasmablasts in vitro similar to in vivo condition. However, to achieve the best outcome in the differentiation of B cells, we should consider the nature of stimulator, the time of incubation, and the type of stimulators.

Background: Many studies showed that regulatory T cells (Tregs) have immunosuppressive effects on immune responses in transplantation and autoimmune disease. Silymarin (isolated from milk thistle or silybum marianum plant) is a flavolignan complex with anti-inflammatory, hepatoprotective, antioxidant and immunomodulatory activities. Previous studies in our group revealed inhibition effect of silymarin on mammalian target of rapamycin (mTOR) activity in activated T cells. Among immunosuppressive drugs, rapamycin can inhibit mTOR, results in Foxp3 expression, Tregs expansion and conventional T cells inhibition. In this study, the effect of silymarin on in-vitro generation of CD4+Foxp3+ cells, in comparison with rapamycin, was evaluated. Methods: Naïve CD4+ T cells were separated from healthy individuals’ peripheral blood mononuclear cells (PBMCs) and activated with monoclonal antibody anti-CD3 and anti-CD28 for 18 hours in Roswell Park Memorial Institute (RPMI) complete medium. Then, incubation was continued with adding Interlukin-2 (IL-2) and silymarin or its control, dimethyl sulfoxide (DMSO), or cultured in present or absent of rapamycin for 3 days. Cells were harvested and stained with anti-CD4 and anti-FoxP3 antibodies for flow cytometry. Findings: Silymarin increased CD4+Foxp3+ T cells compared with its control, DMSO, and with rapamycin after three days of culture of naïve T cells (P 0.05). Conclusion: Given the importance of replacement less harmful medicine and Tregs role in regulating immune system, silymarin, as aTreg generation drug, can be used in the treatment of autoimmune diseases and even in organ transplantation.

مقدمه: سلول TH17 با تولید IL-17 می‌تواند سبب افزایش تولید دو سایتوکین BAFF و APRIL شود که نقش مهمی در بقا و فعال شدن سلول B، و در نتیجه، پاسخ‌های ایمنی هومورال دارند. نارسایی ایمنی رایج متغیر (Common variable immunodeficiency یا CVID)، یکی از بیماری‌های نقص ایمنی اولیه می‌باشد که با کاهش سطح آنتی‌بادی‌ها (نارسایی ایمنی هومورال) همراه است. فراوانی افراد مبتلا به CVID در جمعیت‌های مختلف در محدوده‌ی 1:10000 تا 1:50000 گزارش شده است. هدف از این مطالعه، مقایسه‌ی میانگین بیان ژن IL-17 در بیماران مبتلا به CVID با افراد سالم بود. روش‌ها: ابتدا 2 میلی‌لیتر خون کامل از 10 بیمار مبتلا به CVID و 10 فرد سالم گرفته و سپس تمام mRNA موجود در این نمونه‌ها جدا شد. پس از ساختن cDNA از روی mRNA، میزان بیان ژن IL-17 را با استفاده از روش Real Time-PCR سنجیدیم. یافته‌ها: میزان بیان ژن IL-17 در بیماران مبتلا به CVID نسبت به افراد شاهد کاهش یافت که البته این کاهش از لحاظ آماری معنی‌دار نبود (195/0 = P). نتیجه‌گیری: در این پژوهش نشان دادیم که میزان بیان ژن IL-17 در خون بیماران مبتلا به CVID نسبت به افراد سالم کاهش داشت؛ این موضوع دلیلی بر نقص تولید این سایتوکین است و می‌تواند دلیل نارسایی پاسخ ایمنی هومورال در این افراد باشد.

Background: MicroRNAs (MiRNAs) are 21-24 nucleotides which have different levels of expression between tumors and normal tissues. In this study, we analyzed the expression level of miR-370 in different stages and grades of human breast cancer tissues compared with adjacent normal tissues. Methods: In this case-control study, the expression of miR-370 on 22 tissue samples (tumor and normal) from patients with breast cancer was investigated via quantitative real-time polymerase chain reaction method. Findings: miR-370 expression showed a significant increase in tumor tissues compared to normal tissues (P = 0.001) as well as in cancer tissues at stage III (P = 0.004) and grade 3 (P = 0.011) than in other stages. Conclusion: According to evaluation of miR-370 expression in different stages and grades and its relation to more advanced tumor stages/grades, it can possibility act as oncomiRNA in breast cancer. It may be suggested as a prognostic biomarker or therapeutic target in breast cancer, too.

Objective: Th17 cells are known to be involved in some types of inflammations and autoimmune disorders. RORC2 is the key transcription factor coordinating Th17 cell differentiation. Thus, blocking RORC2 may be useful in suppressing Th17-dependent inflammatory processes. The aim was to silence RORC2 by specific siRNAs in naïve T cells differentiating to Th17. Time-dependent expression of RORC2 as well as IL-17 and IL-23R were considered before and after RORC2 silencing. Materials and Methods: In this experimental study, naïve CD4+ T cells were isolated from human cord blood samples. Cytokines TGFβ plus IL-6 and IL-23 were used to polarize the naïve T cells to Th17 cells in X-VIVO 15 serum free medium. A mixture of three siRNAs specific for RORC2 was applied for blocking its expression. RORC2, IL-17 and IL-23R mRNA and protein levels were measured using qRT-PCR, ELISA and flow cytometry techniques. Pearson correlation and one-way ANOVA were used for statistical analyses. Results: Significant correlations were obtained in time-dependent analysis of IL-17 and IL-23R expression in relation with RORC2 (R=0.87 and 0.89 respectively, p