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Papers by vida homayouni

Natural Killer Cells - Lessons and Challenges [Working Title]

Recurrent miscarriage is an early pregnancy complication that affects about 1–3% of couples. Ther... more Recurrent miscarriage is an early pregnancy complication that affects about 1–3% of couples. There are specific characteristics of natural killer (NK) cells associated with miscarriage. In patients with recurrent miscarriage, a lack of inhibition of decidua natural killer cells can be observed, which leads to a more activated state and presentation of NK cell dim that is characterized by higher levels of pro-inflammatory cytokines and cytotoxicity effect. In peripheral blood, a dysfunctional cytokine production by natural killer cells has been also reported, with an increase of interferon-γ levels and a decrease of interleukin-4. Accordingly, there are different population of NK cells such as dim and bright. The lack of balance between these populations can lead to miscarriage. Using flow cytometry, we can detect these populations and propose the treatment too.

Innate lymphoid cells and cytokines of the novel

Objective: Th17 cells are known to be involved in some types of inflammations and autoimmune diso... more 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 na-ïve 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 pro-tein levels were measured using qRT-PCR, ELISA and flow cytometry techniques. Pearson correlation and one-way ANOVA were used for statistical analyses. Results: Si...

Advanced Biomedical Research, 2014

Background: Microsatellites or simple sequence repeats are repeating sequences of deoxyribonuclei... more Background: Microsatellites or simple sequence repeats are repeating sequences of deoxyribonucleic acid (DNA). Mutation in mismatch repair (MMR) genes can cause microsatellites instability (MSI) in some tumors. In familial disorder of hereditary non-polyposis colorectal cancer (HNPCC), there is a defect in the mechanism of MMR and clearly defective MMR cause unstable microsatellites. This study has been conducted for investigating the instability of microsatellites in alleles of BAT-26 of MSH2 gene in patients with HNPCC in Isfahan, which is an important prognostic biomarker for the prediction of the treatment outcome. Materials and Methods: DNA extraction from forty HNPCC patients peripheral blood samples were performed by using the DNA extraction kit. The polymerase chain reaction (PCR) reaction to amplify BAT-26 was performed. The PCR products were studied by electrophoresis on agarose gel. Results: The size of specific band was 121 bp out of 40 HNPCC samples and based on the above method, it was shown that 12 cases (30%) demonstrated MSI. Chi-square test showed this difference is statistically significant (P < 0.05). Conclusions: The present study was conducted to evaluate the MSI in HNPCC patients. It was determined that the polymorphisms in BAT-26 of MSH2 gene could detect MSI with high sensitivity. Previous reports as well as our results have shown that the use of BAT-26 alone would be sufficient to identify HNPCC-associated MSH2 gene. Identifying MSI in these genes as a marker for prognosis, according to the present study and other researches is important to predict the treatment outcomes.

Background & Aims: Multiple sclerosis (MS) is one of the chronic autoimmune diseases of the centr... more Background & Aims: Multiple sclerosis (MS) is one of the chronic autoimmune diseases of the central nervous system with unknown etiology. The present study aimed to investigate the apoptosis and nitric oxide (NO) production of endothelial cells treated with serum of patients with MS and response to interferon beta (IFN-) therapy. Methods: Human umbilical vein endothelial cells were treated with sera from patients with active MS (in relapse), MS in remission, or sera from healthy volunteers (each n = 10). Nitric oxide (NO) levels were determined in culture supernatants by Greiss method and endothelial cell apoptosis was assessed by annexin V-propidium iodide staining. Effects of IFN-beta-1b on endothelial cell apoptosis and NO production were tested at increasing doses (10, 100, and 1000 U/ml). Results: Compared with healthy people, only apoptosis of endothelial cells treated with serum of patients with relapsing phase increased, P<0.01; while there was no significant difference between apoptosis of endothelial cells treated with serum of patients in remission phase and healthy controls. Apoptosis of endothelial cells treated with sera of patients in relapse was decreased by IFN-beta-1b at 10 U/ml, P<0.05. The same dose also led to a significant increase in nitric oxide production. Conclusion: The results suggest that endothelial cells injury and apoptosis may play a role in MS etiology and represents a potential therapeutic mechanism of action for IFN-beta-1b in MS therapy.

Frontiers in Bioscience, 2012

Iranian Journal of Allergy, Asthma and Immunology

Autoinflammatory diseases (AIDs) are disorders with an inborn error of innate immunity, character... more Autoinflammatory diseases (AIDs) are disorders with an inborn error of innate immunity, characterized by recurrent episodes of fever and inflammatory attacks. The spectrum of AIDs is expanding, but there are no standardized clinical criteria for the diagnosis of the patients. This study aims at establishing the first autoinflammatory registry of an Iranian population focusing on the clinical and laboratory features that may help clinicians for a better understanding and diagnosis of these disorders. Clinical and laboratory characteristics of patients who were clinically and or genetically diagnosed with AIDs were collected for 15 years. The updated version of classification criteria from the Eurofever Registry was used for the clinical diagnosis. Twenty-eight patients (16 males and 12 females) with a mean±SD age of 28.03±14.49 years (from 2 to 68 years) entered this study. About 29% were genetically diagnosed. Familial Mediterranean fever (FMF) was the most common diagnosis of the p...

Composites Science and Technology, 2010

This paper presents the synthesis and characterization of Titanium oxide-poly vinyl alcohol (PVA)... more This paper presents the synthesis and characterization of Titanium oxide-poly vinyl alcohol (PVA) composite ion exchange membrane. The membrane consists of PVA as the base and titanium oxide, with different concentration, as the ionic conductor. The synthesised membranes were analysed on the basis of intermolecular interactions with the help of FT-IR spectroscopy. Characterization techniques such as SEM and TGA were carried out to analyse surface morphology and thermal properties of the synthesised membranes respectively. Transport number was obtained with the help of two compartment diffusion cell. Important parameters such as the water uptake and methanol uptake were also determined. The results were then compared to those available for a well-known commercial membrane called Nafion R. With the increase in the concentration of ionic conducting material from 0.1 g to 0.3 g in 100 ml of crosslinked-PVA solution, the transport number of the membrane increases from 0.86 to 0.89. Increase in the concentration of ionic conductor shows an insignificant effect on other properties like water uptake and methanol uptake and can be safely ignored. The transport number of the synthesized membranes were close to the transport number of Nafion, while the composite membrane performed better when it came to water uptake and methanol uptake as compared to Nafion.

مجله دانشکده پزشکی اصفهان, 2015

Background: T-cell immunoglobulin and mucin domain-3 (Tim3) is known as a marker of cell surface ... more Background: T-cell immunoglobulin and mucin domain-3 (Tim3) is known as a marker of cell surface of T-helper-1 (Th1) cell and has a key role in many diseases with cell-mediated immunity. Over expression of Tim3 has reported in autoimmune and atopic diseases. Though, many studies done about this inhibitory help protein molecules, but yet need more research. In research project on Tim3, native protein with proper post translation modification should be provided. In this study, Tim3 was expressed on the surface of HEK293 T-cell line. Methods: Tim3 expression cassette was amplified from cDNA clone EX-W2682-M67 via polymerase chain reaction (PCR). The PCR product and pHygro plasmid were digested by NheI and MluI restriction enzymes. The linearized pHygro and digested PCR product were ligated together with T4 DNA ligase and transformed into Escherichia coli TOP 10 F'. The resulted pH-Tim3 plasmid was linearized with NheI and transfected into 293T cell line. The transfected cells were ...

Background: RNA interference-based gene silencing has recently been applied as an efficient tool ... more Background: RNA interference-based gene silencing has recently been applied as an efficient tool for functional gene analysis. RORC2 is the key transcription factor orchestrating Th17 cells differentiation, the cells that are known as the pathogenic elements in various autoimmune diseases. The aim of this study was to design efficient siRNAs specific for RORC2 and to evaluate different criteria affecting their functionality. Methods: Three siRNA duplexes specific for RORC2 mRNA were designed. Th17 cells were produced from IL-6 and IL-1 treated cord blood CD4 + T cells. The T cells were transfected with three different designed siRNAs against RORC2 and the expression of RORC2 gene was measured using quantitative real time PCR. Results: Different levels of RORC2 down regulation were observed in the presence of each of the designed siRNAs. Efficient siRNA with 91.1% silencing activity met the majority of the established bioinformatics criteria while the one with 46.6% silencing activit...

Objective: Th17 cells are known to be involved in some types of inflammations and autoimmune diso... more 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: Sig...

BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specif... more BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specific markers of endothelial dysfunction. This study investigated whether CECs and E-selectin are surrogate biomarkers of preeclampsia and if measurement of CECs and E-selectin, early in the third trimester, could be a means of predicting preeclampsia.

Iranian Biomedical Journal, 2008

When fetal red cells enter the maternal circulation from placenta, an event would be happened tha... more When fetal red cells enter the maternal circulation from placenta, an event would be happened that is described as feto-maternal hemorrhage (FMH). This life-threatening condition could be detected by using RBC antigens (surface antigens and intracellular antigens). Therefore, the measurement of fetal RBC in an artificial model would be useful to calculate FMH and consequently the dosage of Rhogam for prophylaxis. The aim of the present study was to evaluate FMH in an artificial mixture model. A series of 40 artificial specimens were prepared consisting of Rh(D) negative adult blood (non-immunized) spiked with varying amounts of Rh(D) positive cord blood from mothers between 20-30 years old in Shahid Beheshti Hospital, Tehran, Iran. Monoclonal anti-D and anti-HbF (fetal hemoglobin) were used for detection of fetal RBC in artificial mixture sample modeling. This study showed that the percentage of fetal cells in artificial sample for anti-D antigen is in ranges of 0.28%-0.32% for a 0....

Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences, 2014

Background: Common variable immunodeficiency (CVID) is characterized by a deficiency in the immun... more Background: Common variable immunodeficiency (CVID) is characterized by a deficiency in the immune system with a heterogeneous collection of disorders resulting in antibody deficiency and recurrent infections. T helper 17 (Th17) cells promote B-cell survival and synergize with the B-cell activating factor to induce their differentiation into the plasma cells. A sub-population of innate lymphoid cells (ILCs) also produces interleukin 17 (IL-17). This study aimed to measure the Th17 specific genes and ILCs counts in the CVID patients in comparison with control subjects. Materials and Methods: Total messenger ribonucleic acid (mRNA) was extracted from the whole blood samples of 10 CVID patients and 10 healthy individuals. IL-17, retinoic acid receptor-related orphan receptor C2 (RORC2), IL-23R, and IL-9 gene expression were measured using the quantitative reverse transcriptase-polymerase chain reaction. Count of lineage negative/CD127+/CD90+ ILCs in the blood samples was performed by t...

Cell Journal (Yakhteh), 2014

Objective Th17 cells are known to be involved in some types of inflammations and autoimmune disor... more 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 TGFß 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 Signific...

Razi Journal of Medical Sciences, 2018

Background: Monocytes are a key component of the immune system, which play an important role in i... more Background: Monocytes are a key component of the immune system, which play an important role in inflammatory and innate immune responses. So the isolation of these cells is important in immunology researches and accordingly many methods have been developed for the monocyte isolation. One of the least expensive and simple separation methods is using the ability of these cells to adhere to solid surfaces that were examined in this study. Methods: In this way, after the isolation of peripheral blood mononuclear cells (PBMCs), monocytes were isolated from cell suspension by using the adhesion ability of these cells to the plastic surface.Isolated monocytes were stained by using specific antibodies labeled with fluorescent dyes (PE, FITC) binding to specific markers on the surface of monocytes CD11b and CD14 and the purity of cells were characterized by flow cytometry. Results: Flow cytometry results after separation of monocytes shown that 90.2% of cells were expressed two markers CD11b...

Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences, 2012

BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specif... more BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specific markers of endothelial dysfunction. This study investigated whether CECs and E-selectin are surrogate biomarkers of preeclampsia and if measurement of CECs and E-selectin, early in the third trimester, could be a means of predicting preeclampsia. METHODS: In this prospective, descriptive-analytic study, rollover test was performed on 523 pregnant women during 28-30 weeks of gestation. CECs were measured by anti-CD 146-driven immunomagnetic isolation in women with positive rollover test. They were followed up prospectively until delivery without any active intervention. Women with and without preeclampsia were determined. The number of CECs and level of E-selectin were compared in the two studied groups. RESULTS: From the 47 pregnant women with positive rollover test who were selected and followed up, 22 individuals were diagnosed with preeclampsia while the remainder were normotensive...

BACKGROUND When fetal red cells enter the maternal circulation from placenta, an event would be h... more BACKGROUND When fetal red cells enter the maternal circulation from placenta, an event would be happened that is described as feto-maternal hemorrhage (FMH). This life-threatening condition could be detected by using RBC antigens (surface antigens and intracellular antigens). Therefore, the measurement of fetal RBC in an artificial model would be useful to calculate FMH and consequently the dosage of Rhogam for prophylaxis. The aim of the present study was to evaluate FMH in an artificial mixture model. METHODS A series of 40 artificial specimens were prepared consisting of Rh(D) negative adult blood (non-immunized) spiked with varying amounts of Rh(D) positive cord blood from mothers between 20-30 years old in Shahid Beheshti Hospital, Tehran, Iran. Monoclonal anti-D and anti-HbF (fetal hemoglobin) were used for detection of fetal RBC in artificial mixture sample modeling. RESULTS This study showed that the percentage of fetal cells in artificial sample for anti-D antigen is in ran...

Iranian Journal of Biotechnology

Multiple Sclerosis and Related Disorders

Natural Killer Cells - Lessons and Challenges [Working Title]

Recurrent miscarriage is an early pregnancy complication that affects about 1–3% of couples. Ther... more Recurrent miscarriage is an early pregnancy complication that affects about 1–3% of couples. There are specific characteristics of natural killer (NK) cells associated with miscarriage. In patients with recurrent miscarriage, a lack of inhibition of decidua natural killer cells can be observed, which leads to a more activated state and presentation of NK cell dim that is characterized by higher levels of pro-inflammatory cytokines and cytotoxicity effect. In peripheral blood, a dysfunctional cytokine production by natural killer cells has been also reported, with an increase of interferon-γ levels and a decrease of interleukin-4. Accordingly, there are different population of NK cells such as dim and bright. The lack of balance between these populations can lead to miscarriage. Using flow cytometry, we can detect these populations and propose the treatment too.

Innate lymphoid cells and cytokines of the novel

Objective: Th17 cells are known to be involved in some types of inflammations and autoimmune diso... more 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 na-ïve 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 pro-tein levels were measured using qRT-PCR, ELISA and flow cytometry techniques. Pearson correlation and one-way ANOVA were used for statistical analyses. Results: Si...

Advanced Biomedical Research, 2014

Background: Microsatellites or simple sequence repeats are repeating sequences of deoxyribonuclei... more Background: Microsatellites or simple sequence repeats are repeating sequences of deoxyribonucleic acid (DNA). Mutation in mismatch repair (MMR) genes can cause microsatellites instability (MSI) in some tumors. In familial disorder of hereditary non-polyposis colorectal cancer (HNPCC), there is a defect in the mechanism of MMR and clearly defective MMR cause unstable microsatellites. This study has been conducted for investigating the instability of microsatellites in alleles of BAT-26 of MSH2 gene in patients with HNPCC in Isfahan, which is an important prognostic biomarker for the prediction of the treatment outcome. Materials and Methods: DNA extraction from forty HNPCC patients peripheral blood samples were performed by using the DNA extraction kit. The polymerase chain reaction (PCR) reaction to amplify BAT-26 was performed. The PCR products were studied by electrophoresis on agarose gel. Results: The size of specific band was 121 bp out of 40 HNPCC samples and based on the above method, it was shown that 12 cases (30%) demonstrated MSI. Chi-square test showed this difference is statistically significant (P < 0.05). Conclusions: The present study was conducted to evaluate the MSI in HNPCC patients. It was determined that the polymorphisms in BAT-26 of MSH2 gene could detect MSI with high sensitivity. Previous reports as well as our results have shown that the use of BAT-26 alone would be sufficient to identify HNPCC-associated MSH2 gene. Identifying MSI in these genes as a marker for prognosis, according to the present study and other researches is important to predict the treatment outcomes.

Background & Aims: Multiple sclerosis (MS) is one of the chronic autoimmune diseases of the centr... more Background & Aims: Multiple sclerosis (MS) is one of the chronic autoimmune diseases of the central nervous system with unknown etiology. The present study aimed to investigate the apoptosis and nitric oxide (NO) production of endothelial cells treated with serum of patients with MS and response to interferon beta (IFN-) therapy. Methods: Human umbilical vein endothelial cells were treated with sera from patients with active MS (in relapse), MS in remission, or sera from healthy volunteers (each n = 10). Nitric oxide (NO) levels were determined in culture supernatants by Greiss method and endothelial cell apoptosis was assessed by annexin V-propidium iodide staining. Effects of IFN-beta-1b on endothelial cell apoptosis and NO production were tested at increasing doses (10, 100, and 1000 U/ml). Results: Compared with healthy people, only apoptosis of endothelial cells treated with serum of patients with relapsing phase increased, P<0.01; while there was no significant difference between apoptosis of endothelial cells treated with serum of patients in remission phase and healthy controls. Apoptosis of endothelial cells treated with sera of patients in relapse was decreased by IFN-beta-1b at 10 U/ml, P<0.05. The same dose also led to a significant increase in nitric oxide production. Conclusion: The results suggest that endothelial cells injury and apoptosis may play a role in MS etiology and represents a potential therapeutic mechanism of action for IFN-beta-1b in MS therapy.

Frontiers in Bioscience, 2012

Iranian Journal of Allergy, Asthma and Immunology

Autoinflammatory diseases (AIDs) are disorders with an inborn error of innate immunity, character... more Autoinflammatory diseases (AIDs) are disorders with an inborn error of innate immunity, characterized by recurrent episodes of fever and inflammatory attacks. The spectrum of AIDs is expanding, but there are no standardized clinical criteria for the diagnosis of the patients. This study aims at establishing the first autoinflammatory registry of an Iranian population focusing on the clinical and laboratory features that may help clinicians for a better understanding and diagnosis of these disorders. Clinical and laboratory characteristics of patients who were clinically and or genetically diagnosed with AIDs were collected for 15 years. The updated version of classification criteria from the Eurofever Registry was used for the clinical diagnosis. Twenty-eight patients (16 males and 12 females) with a mean±SD age of 28.03±14.49 years (from 2 to 68 years) entered this study. About 29% were genetically diagnosed. Familial Mediterranean fever (FMF) was the most common diagnosis of the p...

Composites Science and Technology, 2010

This paper presents the synthesis and characterization of Titanium oxide-poly vinyl alcohol (PVA)... more This paper presents the synthesis and characterization of Titanium oxide-poly vinyl alcohol (PVA) composite ion exchange membrane. The membrane consists of PVA as the base and titanium oxide, with different concentration, as the ionic conductor. The synthesised membranes were analysed on the basis of intermolecular interactions with the help of FT-IR spectroscopy. Characterization techniques such as SEM and TGA were carried out to analyse surface morphology and thermal properties of the synthesised membranes respectively. Transport number was obtained with the help of two compartment diffusion cell. Important parameters such as the water uptake and methanol uptake were also determined. The results were then compared to those available for a well-known commercial membrane called Nafion R. With the increase in the concentration of ionic conducting material from 0.1 g to 0.3 g in 100 ml of crosslinked-PVA solution, the transport number of the membrane increases from 0.86 to 0.89. Increase in the concentration of ionic conductor shows an insignificant effect on other properties like water uptake and methanol uptake and can be safely ignored. The transport number of the synthesized membranes were close to the transport number of Nafion, while the composite membrane performed better when it came to water uptake and methanol uptake as compared to Nafion.

مجله دانشکده پزشکی اصفهان, 2015

Background: T-cell immunoglobulin and mucin domain-3 (Tim3) is known as a marker of cell surface ... more Background: T-cell immunoglobulin and mucin domain-3 (Tim3) is known as a marker of cell surface of T-helper-1 (Th1) cell and has a key role in many diseases with cell-mediated immunity. Over expression of Tim3 has reported in autoimmune and atopic diseases. Though, many studies done about this inhibitory help protein molecules, but yet need more research. In research project on Tim3, native protein with proper post translation modification should be provided. In this study, Tim3 was expressed on the surface of HEK293 T-cell line. Methods: Tim3 expression cassette was amplified from cDNA clone EX-W2682-M67 via polymerase chain reaction (PCR). The PCR product and pHygro plasmid were digested by NheI and MluI restriction enzymes. The linearized pHygro and digested PCR product were ligated together with T4 DNA ligase and transformed into Escherichia coli TOP 10 F'. The resulted pH-Tim3 plasmid was linearized with NheI and transfected into 293T cell line. The transfected cells were ...

Background: RNA interference-based gene silencing has recently been applied as an efficient tool ... more Background: RNA interference-based gene silencing has recently been applied as an efficient tool for functional gene analysis. RORC2 is the key transcription factor orchestrating Th17 cells differentiation, the cells that are known as the pathogenic elements in various autoimmune diseases. The aim of this study was to design efficient siRNAs specific for RORC2 and to evaluate different criteria affecting their functionality. Methods: Three siRNA duplexes specific for RORC2 mRNA were designed. Th17 cells were produced from IL-6 and IL-1 treated cord blood CD4 + T cells. The T cells were transfected with three different designed siRNAs against RORC2 and the expression of RORC2 gene was measured using quantitative real time PCR. Results: Different levels of RORC2 down regulation were observed in the presence of each of the designed siRNAs. Efficient siRNA with 91.1% silencing activity met the majority of the established bioinformatics criteria while the one with 46.6% silencing activit...

Objective: Th17 cells are known to be involved in some types of inflammations and autoimmune diso... more 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: Sig...

BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specif... more BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specific markers of endothelial dysfunction. This study investigated whether CECs and E-selectin are surrogate biomarkers of preeclampsia and if measurement of CECs and E-selectin, early in the third trimester, could be a means of predicting preeclampsia.

Iranian Biomedical Journal, 2008

When fetal red cells enter the maternal circulation from placenta, an event would be happened tha... more When fetal red cells enter the maternal circulation from placenta, an event would be happened that is described as feto-maternal hemorrhage (FMH). This life-threatening condition could be detected by using RBC antigens (surface antigens and intracellular antigens). Therefore, the measurement of fetal RBC in an artificial model would be useful to calculate FMH and consequently the dosage of Rhogam for prophylaxis. The aim of the present study was to evaluate FMH in an artificial mixture model. A series of 40 artificial specimens were prepared consisting of Rh(D) negative adult blood (non-immunized) spiked with varying amounts of Rh(D) positive cord blood from mothers between 20-30 years old in Shahid Beheshti Hospital, Tehran, Iran. Monoclonal anti-D and anti-HbF (fetal hemoglobin) were used for detection of fetal RBC in artificial mixture sample modeling. This study showed that the percentage of fetal cells in artificial sample for anti-D antigen is in ranges of 0.28%-0.32% for a 0....

Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences, 2014

Background: Common variable immunodeficiency (CVID) is characterized by a deficiency in the immun... more Background: Common variable immunodeficiency (CVID) is characterized by a deficiency in the immune system with a heterogeneous collection of disorders resulting in antibody deficiency and recurrent infections. T helper 17 (Th17) cells promote B-cell survival and synergize with the B-cell activating factor to induce their differentiation into the plasma cells. A sub-population of innate lymphoid cells (ILCs) also produces interleukin 17 (IL-17). This study aimed to measure the Th17 specific genes and ILCs counts in the CVID patients in comparison with control subjects. Materials and Methods: Total messenger ribonucleic acid (mRNA) was extracted from the whole blood samples of 10 CVID patients and 10 healthy individuals. IL-17, retinoic acid receptor-related orphan receptor C2 (RORC2), IL-23R, and IL-9 gene expression were measured using the quantitative reverse transcriptase-polymerase chain reaction. Count of lineage negative/CD127+/CD90+ ILCs in the blood samples was performed by t...

Cell Journal (Yakhteh), 2014

Objective Th17 cells are known to be involved in some types of inflammations and autoimmune disor... more 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 TGFß 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 Signific...

Razi Journal of Medical Sciences, 2018

Background: Monocytes are a key component of the immune system, which play an important role in i... more Background: Monocytes are a key component of the immune system, which play an important role in inflammatory and innate immune responses. So the isolation of these cells is important in immunology researches and accordingly many methods have been developed for the monocyte isolation. One of the least expensive and simple separation methods is using the ability of these cells to adhere to solid surfaces that were examined in this study. Methods: In this way, after the isolation of peripheral blood mononuclear cells (PBMCs), monocytes were isolated from cell suspension by using the adhesion ability of these cells to the plastic surface.Isolated monocytes were stained by using specific antibodies labeled with fluorescent dyes (PE, FITC) binding to specific markers on the surface of monocytes CD11b and CD14 and the purity of cells were characterized by flow cytometry. Results: Flow cytometry results after separation of monocytes shown that 90.2% of cells were expressed two markers CD11b...

Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences, 2012

BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specif... more BACKGROUND: Circulating endothelial cells (CECs) and E-selectin are known as sensitive and specific markers of endothelial dysfunction. This study investigated whether CECs and E-selectin are surrogate biomarkers of preeclampsia and if measurement of CECs and E-selectin, early in the third trimester, could be a means of predicting preeclampsia. METHODS: In this prospective, descriptive-analytic study, rollover test was performed on 523 pregnant women during 28-30 weeks of gestation. CECs were measured by anti-CD 146-driven immunomagnetic isolation in women with positive rollover test. They were followed up prospectively until delivery without any active intervention. Women with and without preeclampsia were determined. The number of CECs and level of E-selectin were compared in the two studied groups. RESULTS: From the 47 pregnant women with positive rollover test who were selected and followed up, 22 individuals were diagnosed with preeclampsia while the remainder were normotensive...

BACKGROUND When fetal red cells enter the maternal circulation from placenta, an event would be h... more BACKGROUND When fetal red cells enter the maternal circulation from placenta, an event would be happened that is described as feto-maternal hemorrhage (FMH). This life-threatening condition could be detected by using RBC antigens (surface antigens and intracellular antigens). Therefore, the measurement of fetal RBC in an artificial model would be useful to calculate FMH and consequently the dosage of Rhogam for prophylaxis. The aim of the present study was to evaluate FMH in an artificial mixture model. METHODS A series of 40 artificial specimens were prepared consisting of Rh(D) negative adult blood (non-immunized) spiked with varying amounts of Rh(D) positive cord blood from mothers between 20-30 years old in Shahid Beheshti Hospital, Tehran, Iran. Monoclonal anti-D and anti-HbF (fetal hemoglobin) were used for detection of fetal RBC in artificial mixture sample modeling. RESULTS This study showed that the percentage of fetal cells in artificial sample for anti-D antigen is in ran...

Iranian Journal of Biotechnology

Multiple Sclerosis and Related Disorders