Eric Adua; Frank Oteng Danso; Oswald Mensah Boa-Amponsem; Frank Adusei-Mensah
Volume 12, Issue 2 , June 2015, , Pages 94-103
Abstract
Background: During the initial phase of an infection, there is an upregulation of inducible nitric oxide synthase in the macrophages for the production of nitric oxide. This is followed by the recruitment of polymorphonuclear leukocytes (neutrophils) which release arginase. Arginase competes with inducible ...
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Background: During the initial phase of an infection, there is an upregulation of inducible nitric oxide synthase in the macrophages for the production of nitric oxide. This is followed by the recruitment of polymorphonuclear leukocytes (neutrophils) which release arginase. Arginase competes with inducible nitric oxide synthase for a common substrate L-arginine. Objective: To investigate whether the entry of neutrophils and release of arginase can interfere with nitric oxide production from stimulated mouse macrophages. Methods: Neutrophils were isolated from human blood and stimulated with cytodex-3 beads. Cultured macrophages were stimulated with lipopolysaccharide and interferon gamma with or without N (G)-nitro-L-arginine methyl ester or N (omega)-hydroxy-nor-L-arginine. Measurement of NO2 - /NO3 - and urea were done using the spectrophotometer. Results: A significantly higher level of nitric oxide production from stimulated macrophages was observed compared to control. There was a decrease in nitric oxide production when stimulated macrophages were treated with the supernatant from activated neutrophils (p<0.05). Conclusion: Arginase from neutrophils can modulate nitric oxide production from activated macrophages which may affect the course of infection by intracellular bacteria.
mahdi aminikhah; Mir Saeed Yekaninejad; M.Hosein Nicknam; Farideh Khosravi; Mehrnaz naroei nejad; Bita Ansaripour; batol moradi; Behrouz Nikbin; Aliakbar Amirzargar
Abstract
Background: The high polymorphism in the human leukocyte antigen (HLA) genes can be used as an identity of individuals to compare with other populations. This extreme polymorphism in the HLA system is accountable for the differences in alleles and haplotypes among ethnic groups, populations, and the ...
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Background: The high polymorphism in the human leukocyte antigen (HLA) genes can be used as an identity of individuals to compare with other populations. This extreme polymorphism in the HLA system is accountable for the differences in alleles and haplotypes among ethnic groups, populations, and the inhabitants of many regions. Objective: To define the frequency of HLA alleles and haplotypes among the Sistanis, Sistani/Zaboli population in Iran. Methods: In this study, genotyping of class I (A, B, C) and class II HLA (DRB1, DQA1, DQB1) loci were determined in 90 unrelated Iraninan Sistani people and the results were compared with 474,892 HLA chromosomes from a diverse worldwide population. Results: The highest frequently observed alleles in this study were A*02:01, B*35:01, C*12:03, C*06:02, DRB1*11, DQA1*05:05, and DQB1*03:01. Furthermore, the most frequent 3-locus haplotypes were A*02:01-B*50:01*C*06:02, DRB1*11-DQB1*03:01-DQA1*05:05, and A*02:01-B*50:01-DRB1*07. The most occurring 4-locus haplotypes were A*02:01-B*50:01-C*06:02-DRB1*07 and A*02:01-B*50:01-DRB1*07-DQB1*02:01. A*02:01-B*50:01-C*06:02-DRB1*07-DQB1*02:01 and A*02:01-B*50:01-C*06:02-DRB1*07-DQB1*02:01-DQA1*02:01 were determined to be the predominant 5- and 6-locus haplotypes, respectively. The heat maps and multiple correspondence analyses based on the frequency of HLA alleles showed that Sistanis share a common genetic inheritance with other Iranian ethnic groups such as the people from Yazd and Fars except some differences with Baluchis, Iranian Jews, Lurs of Kohgiluyeh/Buyerahmad, and Arabs of Fars, which may arise from the admixture of these groups or with foreign subgroups over centuries, and also a close relatedness with some European populations. Conclusion: These data could be useful for finding better donor matches for organ transplantation among Sistanis or other related Iranian ethnic groups, epidemiological studies of HLA-associated diseases, handling HLA genomics and mapping the migration pattern of different ethnic group.
Abdollah Jafarzadeh; Jalal Khoshnoodi; Shayesteh Ghorbani; Saleh Mohaghegh Hazrati; Babak Faraj Mazaheri; Fazel Shokri
Volume 1, Issue 2 , September 2004, , Pages 98-104
Abstract
Objective: To compare immunogenicity of a recombinant hepatitis B (HB) vaccine in two groups of neonates born in two cities of Iran with different geographic and ethnic backgrounds. Materials and Methods: Ten micrograms of a recombinant HB vaccine was administered under field condition to Iranian ...
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Objective: To compare immunogenicity of a recombinant hepatitis B (HB) vaccine in two groups of neonates born in two cities of Iran with different geographic and ethnic backgrounds. Materials and Methods: Ten micrograms of a recombinant HB vaccine was administered under field condition to Iranian healthy neonates at 0, 1.5 and 9 months intervals. The subjects consisted of two groups of 290 and 231 neonates selected from two cities located at north-west (Urmia) and south-east (Kerman) of Iran, respectively. The level of anti-HBs antibody was quantitated in serum 2-4 weeks after administration of the last vaccine dose, by sandwich ELISA. Results: A higher seroprotection rate (anti-HBs> 10 IU/L) (98.3% vs. 96.1%) and significantly increased serum anti- HBs antibody titer (11869 vs. 6104 IU/L) (P<0.001) were induced in vaccinated neonates from Urmia city, compared to those born in Kerman. Conclusion: These findings suggest contribution of ethnic and/or environmental factors in the antibody response to recombinant HB vaccine in human.
Fateme Sadri-Ardalani; Moslem Ahmadi; Azam Hemmati; Shaghayegh Emami; Samira Farid; Mohammad Mehdi Amiri; Mahmood Jeddi-Tehrani; Mahdi Shabani; Fazel Shokri
Volume 14, Issue 2 , June 2017, , Pages 99-110
Abstract
Background: In addition to passive immunotherapy using anti-HER2 monoclonal antibodies, active immunotherapy via HER2 targeting is an interesting approach to inducing specific anti-tumor immune responses. We have recently reported the immunogenicity of HER2 subdomains following DNA immunization and HER2 ...
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Background: In addition to passive immunotherapy using anti-HER2 monoclonal antibodies, active immunotherapy via HER2 targeting is an interesting approach to inducing specific anti-tumor immune responses. We have recently reported the immunogenicity of HER2 subdomains following DNA immunization and HER2 protein boosting. In the present study, we evaluated the immunogenicity of different HER2 extracellular subdomains for the induction of anti-HER2 antibody response in BALB/c mice. Objective: To investigate and characterize antibody responses to human recombinant proteins of HER2 extracellular subdomains in immunized mice. Methods: Four subdomains of HER2 extracellular domain were expressed in E.coli; subsequently, purified recombinant proteins were intraperitoneally injected in BALB/c mice with Freund's adjuvant. The anti-HER2 antibody response was detected by ELISA, immunoblotting and flow cytometry. Results: All the four HER2 subdomains along with the full extracellular domain (fECD) were able to induce specific anti-HER2 antibodies. Although anti-HER2 subdomains antibodies could not react with eukaryotic recombinant fECD protein by ELISA, they were able to recognize this protein by immunoblotting under both reduced and non-reduced conditions. Furthermore, only the sera of mice immunized with fECD protein could recognize native HER2 on HER2 overexpressing tumor cells (>99%) by flow cytometry. Moreover, fECD immunized mice sera inhibited the proliferation of tumor cells by XTT assay. Conclusion: The prokaryotic recombinant proteins of HER2 extracellular subdomains are immunogenic, yet the induced specific antibodies do not react with the native HER2 protein due to the paucity of post-translation modifications and /or distortion of the native conformation of isolated HER2 extracellular subdomains which might be potentially effective for induction of cell mediated immune response against HER2.
Muneo Numasaki; Koyu Ito
Abstract
Background: Interleukin (IL)-17A possesses biological activities to promote vascular endothelial cell migration and microvessel development. Objective: To clarify which angiogenic factors are involved in IL-17A-modified angiogenesis-related functions of vascular endothelial cell migration and microtube ...
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Background: Interleukin (IL)-17A possesses biological activities to promote vascular endothelial cell migration and microvessel development. Objective: To clarify which angiogenic factors are involved in IL-17A-modified angiogenesis-related functions of vascular endothelial cell migration and microtube development or not. Methods: The potential contribution of various angiogenic stimulators to in vitro angiogenic activities of IL-17A was assessed with both modified Boyden Chemotaxicell chamber assay and in vitro angiogenesis assay. Results: The addition of a neutralizing antibody (Ab) for hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF)-A to the upper and lower compartments in a modified Boyden Chemotaxicell chamber significantly attenuated human dermal microvascular endothelial cell (HMVEC) migration elicited by IL-17A. Moreover, IL-17A-induced capillary-like microvessel development in human umbilical vein endothelial cell (HUVEC) and human dermal fibroblast (HDF) co-culture system was significantly impaired by a neutralizing Ab against HGF, bFGF, VEGF-A, cysteine-x-cysteine ligand 8 (CXCL8)/IL-8 or cysteine-x-cysteine (CXC) chemokine receptor (CXCR)-2. Conclusion: Our findings demonstrate the involvement of HGF, bFGF, VEGF-A and/or CXCL8/IL-8, to various degrees, in migration and microvessel development of vascular endothelial cells mediated by IL-17A.
Shirin Farjadian; Abbas Ghaderi
Volume 3, Issue 3 , September 2006, , Pages 106-113
Abstract
Background: HLA genes are highly polymorphic and certain alleles are frequent only in specific populations. Therefore, HLA is a unique tool for studying the genetic relationship between different populations. Iranians are ethnically diverse people and one of the major ethnic groups in Iran is Lur population ...
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Background: HLA genes are highly polymorphic and certain alleles are frequent only in specific populations. Therefore, HLA is a unique tool for studying the genetic relationship between different populations. Iranians are ethnically diverse people and one of the major ethnic groups in Iran is Lur population inhabiting along the central and southern parts of Zagros Chain Mountain. Objectives: Genetic relationship among three Lur subpopulations was investigated based on HLA class II profiles. Methods: HLA typing was performed using PCR/RFLP and PCR/SSP methods in 154 individuals from three Lur subpopulation living in Luristan, Kohkiloyeh/ Boyerahmad, and Chahar-Mahal/ Bakhtiari. Results: The most common DRB1 allele in Lurs of Luristan and Kohkiloyeh/ Boyerahmad was *1103=4 while DRB1*0701 was the most common allele in Bakhtiaris. DQA1*0501 and DQB1*0301 were the most frequent alleles and DRB1*1103=04-DQA1*0501-DQB1*0301 was the predominant haplotype in the three studied subpopulations. Neighbor-joining tree based on Nei's genetic distances and correspondence analysis according to DRB1, DQA1, and DQB1 allele frequencies showed a close genetic relationship between Lurs of Luristan and Lurs of Kohkiloye/ Boyerahmad and they were well separated from Bakhtiaris. The results of AMOVA revealed no significant difference between the three studied groups of Lurs and other major ethnic groups of Iran. Conclusion: The results of this study revealed that Bakhtiaris were genetically far from the two other Lur subpopulations. Despite a probable common ancestor, this genetic difference might be explained by Bakhtiaris admixture with other Zagros inhabitants due to their nomadic life style.
Mohsen Arabpour; Atri Ghods; Mahmoud Shariat; Abodl-Rasoul Talei; Fereshteh Mehdipour; Abbas Ghaderi
Abstract
Background: B cells can increase the expression of granzyme B in CD8+ T cells through 4-1BBL/4-1BB interaction and promote anti-tumor immunity. Objective: To investigate the expression of 4-1BBL on B cells in the breast tumor draining lymph nodes (TDLNs) and its association with disease parameters. Methods: ...
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Background: B cells can increase the expression of granzyme B in CD8+ T cells through 4-1BBL/4-1BB interaction and promote anti-tumor immunity. Objective: To investigate the expression of 4-1BBL on B cells in the breast tumor draining lymph nodes (TDLNs) and its association with disease parameters. Methods: Using Ficoll-Hypaque gradient centrifugation, mononuclear cells were isolated from axillary lymph nodes of 42 patients. Cells received 4 hours of PMA/Ionomycin stimulation, in vitro. Both unstimulated and stimulated cells were stained with anti‒CD19 and anti‒4-1BBL antibodies and subjected to flow cytometry. Results: 4-1BBL expression was detected on 2.8 ± 1.7% of unstimulated B cells, while 27.4 ± 11.9% of B cells expressed this co-stimulatory molecule following stimulation. In steady state, the percentage of 4-1BBL+ B cells was not associated with cancer characteristics. However, in patients with invasive ductal carcinoma, the percentage of 4-1BBL expressing B cells in stimulated condition had a decreasing trend in grade III, compared to grade II+I. In addition, significantly higher frequency of 4-1BBL+ B cells was seen in the TDLNs of ER+ or PR+ compared with ER‒ or PR‒ patients (p=0.021 and p=0.015, respectively). No significant associations were observed between the frequency of 4-1BBL+ B cells and the number of involved LNs, Her2 expression or disease stage. Conclusions: The frequency of 4-1BBL+ B cells significantly increased following a short time activation, and showed relative and significant associations with tumor grade and estrogen receptor status, respectively. More investigations are required to evaluate the potential of 4-1BBL+ B cells for use in immunotherapy.
Hossein Forghani; Mahin Jamshidi Makiani; Hossein Zarei Jaliani; Seyed Mohsen Zahraei; Seyedeh Mahdieh Namayandeh; Parisa Khani
Abstract
Background: Currently evidence indicates the resurgence of whooping cough despite high coverage of whole-cell (wP) and acellular (aP) pertussis vaccines. Objective: In this study, we investigated the cell-mediated immune response of a genetically inactivated protein containing the S1 subunit of pertussis ...
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Background: Currently evidence indicates the resurgence of whooping cough despite high coverage of whole-cell (wP) and acellular (aP) pertussis vaccines. Objective: In this study, we investigated the cell-mediated immune response of a genetically inactivated protein containing the S1 subunit of pertussis toxin (PTS1) without and with the Listeriolysin O (LLO-PTS1), developed by the researchers (the authors of this study), in comparison with current wP and aP vaccines in the mice model. Methods: Thirty-six female NMRI mice aged 8 to 12 weeks (25 ± 5 g) were divided into six groups including control (n=6), and five treated groups (n=6/each). Treated groups comprising recombinant PTS1, recombinant fusion LLO-PTS1, aP, wP, and sham (phosphate-buffered saline) were injected intraperitoneally whereas the control group did not receive anything. After 60 days, the serum levels of IFN-γ, IL-4, and IL-17 cytokines (as the T-helper 1, 2, and 17 responses, respectively) were evaluated by mouse ELISA Kit. Results: Our findings showed LLO-PTS1 significantly increased IL-17 and IL-4 cytokines compared with wP and aP vaccines (superiority). IFN-γ failed to significantly increase in the LLO-PTS1 group compared to others but it was non-inferior to standard vaccines (non-inferiority). Conclusion: Our alum free mono-component monovalent recombinant fusion protein (LLO-PTS1), registered as a patent in the www.iripo.ssaa.ir, could bear the capacity to stimulate the Th-1 response similar to wP and aP vaccines (non-inferiority) in the mice model. In addition, it showed better results in Th-17 and Th-2 response (superiority). This study can be regarded as a springboard for further probes in booster pertussis vaccine development.
Mojgan Mohammadi; Mohammad Reza Bazrafshani; Philip.J Day; William. E.R. Ollier
Volume 6, Issue 3 , September 2009, , Pages 119-129
Abstract
Background: Vascular endothelial growth factor (VEGF) has a key role in angiogene-sis and in transplantation. The level of VEGF is related to the differences in the DNA sequence of its promoter region. Objectives: In this study, the association between the combination of VEGF –1154 G and –2578 ...
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Background: Vascular endothelial growth factor (VEGF) has a key role in angiogene-sis and in transplantation. The level of VEGF is related to the differences in the DNA sequence of its promoter region. Objectives: In this study, the association between the combination of VEGF –1154 G and –2578 C alleles and VEGF production by LPS-stimulated PBMCs was investigated. In addition; the relationship between VEGF poly-morphisms and the influence of TNF-α and IL-4 on VEGF production was studied. Methods: VEGF –1154 G/A and –2578 C/A were detected using ARMS-PCR. To de-termine the impact of combinations of these two polymorphisms on VEGF production; PBMCs were stimulated by LPS and VEGF production was measured by ELISA. Re-sults: The combinations of –1154 GG/-2578 CC and –1154 GG/-2578 CA were signifi-cantly associated with higher VEGF production (p<0.0001). Production of VEGF was significantly influenced by TNF-α in individuals who had certain VEGF genotype com-binations. Although VEGF production was dramatically suppressed by IL-4, it was not dependent on VEGF genotype. Conclusions: Since TNF-α has influence on the graft outcome, to avoid allocation of grafts from high TNF-α producer donors to recipients, it might be useful to predict and minimize graft rejection by having prior knowledge of TNF-α and also VEGF genotypes especially -1154 G/A and -2578 C/A VEGF.
Fatemeh Vahedi; Naser Taiebi Meibody; Mahdi KianiZadeh; Mahmoud Mahmoudi
Volume 2, Issue 3 , September 2005, , Pages 134-140
Abstract
Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence ...
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Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence of protective antigen of Bacillus anthracis, amino acids 175-764, as a potent immunogenic target was selected. The DNA encoding this segment was utilized in the construction of pcDNA3.1+PA plasmid. After intramuscular injection of rats with pcDNA3.1+PA plasmid, the expression of PA was assessed by RT-PCR and immunohistochemistry at RNA and protein levels, respectively. We also evaluated the presence of anti-PA antibodies in sera of immunized mice with pcDNA3.1+PA construct using immunoblotting. Results: The integrity of pcDNA3.1+PA construct was confirmed with restriction analysis and sequencing. The expression of PA was detected at RNA and protein levels. The presence of anti-PA antibodies in immunized mice with pcDNA3.1+PA construct was also confirmed. Conclusion: Our results indicate that pcDNA3.1+PA eukaryotic expressing vector could express PA antigen, induce antibody response and may be used as a candidate for DNA vaccine against anthrax.
Maryam Azimi Mohamadabadi; Zuhair Mohammad Hassan; Ahmad Zavaran Hosseini; Mehrdad Gholamzad; Shekoofe Noori; Mehdi Mahdavi; Hamidreza Maroof
Volume 10, Issue 3 , September 2013, , Pages 139-149
Abstract
Background: Chemo-immunotherapy is one of the new achievements for treatment of cancer, by which the success of anti-cancer therapy can be increased. In vitro studies have been shown that Arteether (ARE) induces apoptosis in tumor cells, but not in normal cells. Objective: To investigate the cytotoxic ...
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Background: Chemo-immunotherapy is one of the new achievements for treatment of cancer, by which the success of anti-cancer therapy can be increased. In vitro studies have been shown that Arteether (ARE) induces apoptosis in tumor cells, but not in normal cells. Objective: To investigate the cytotoxic and immunomodulatory properties of Arteether in-vivo and in-vitro. Methods: In this study, we used MTT assay for evaluation of cytotoxicity of Arteether on tumor cell line and Peripheral Blood Mononuclear Cells (PBMCs) from healthy individuals. Balb/c mice were subcutaneously transplanted with tumor tissue taken from Spontaneous Mouse Mammary Tumor (SMMT) bearing female mice. Arteether was administered to breast tumor-bearing Balb/c mice at a dose of 6 mg/kg/day intraperitoneally. Tumor sizes, lymphocyte proliferation, cytokines production, and the percentage of splenic T-reg cells were measured. Results: We observed that ARE could reduce the cell growth of 4T1 cell line in a dose-dependent manner but it had no cytotoxic effect on the growth of peripheral blood lymphocytes. ARE administered intraperitoneally to tumor-bearing Balb/c mice could reduce the tumor growth rate and splenic T-reg cells. No difference in the IFN-γ, IL-10 and IL-4 production was observed between tumor antigenstimulated splenocytes of mice treated with ARE and control mice. Conclusion: These results underscore antitumor properties of Arteether that may aid in development of more effective antitumor agents.
Tiantian Cai; Guofei Wang; Yanping Yang; Kaida Mu; Jing Zhang; Yanfei Jiang; Jin-an Zhang
Abstract
Background: Several autoimmune and inflammatory disorders, including autoimmune thyroid diseases (AITD), have been linked to Th17 cells and the IL-23/IL-17 axis. Current data suggest that genetic variation contributes greatly to disease susceptibility to AITD. Objectives: To study the role ...
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Background: Several autoimmune and inflammatory disorders, including autoimmune thyroid diseases (AITD), have been linked to Th17 cells and the IL-23/IL-17 axis. Current data suggest that genetic variation contributes greatly to disease susceptibility to AITD. Objectives: To study the role of single nucleotide polymorphisms (SNPs) of IL-23/IL-17 pathway in AITD predisposition and test the gene-gene/gene-sex interactions in these loci. Methods: A total of 1051 patients with AITD, including 657 patients with Graves' disease (GD) and 394 patients with Hashimoto's thyroiditis (HT), and 874 healthy controls were enrolled in this case-control association study. Six SNPs were selected and genotyped by multiplex PCR combined with high-throughput sequencing. Interactions were tested by the general multifactor dimensionality reduction (GMDR) method. Results: Allele C and combinational genotype AC+CC of rs3212227 within IL-23 were significantly associated with GD with goiter (p=0.003 and 0.014, respectively). Allele G and combinational genotype AG+GG of rs4819554 within IL-17RA were significantly related to HT with family history and the severity of HT (p=0.011 and 0.027; p=0.041 and 0.035). Also, allele T and genotype CT+TT of rs9463772 within IL-17F were significantly correlated with the severity of HT (p=0.001 and 0.027, respectively). Moreover, high dimensional gene-sex interaction (IL-23R-IL-23-IL-17RA-IL-17F-sex) was identified in AITD, GD, and HT patients with GMDR analysis. Conclusions: Our study identified the novel loci and gene-sex interaction in AITD. This evidence, from another perspective, suggests that sex, IL-23/IL-17 pathway, and Th17 cells play an important role in the pathogenesis of AITD.
Marzieh Holakuyee; Mohammad Hossein Yadegari; Zuhair Mohammad Hassan; Mansour Bayat; Ariyo Shahin Jafari; Mohsen Abolhassani; Abbas Ali Amini; Mehdi Mahdavi
Volume 7, Issue 3 , September 2010, , Pages 142-149
Abstract
Background: Candida albicans is one of the most important opportunistic pathogens that suppress immunologic mechanisms of the host. It is speculated that structural and secretory proteins of C. albicans have immunomodulatory effects in cancer. Objective: To evaluate the effects of C. albicans structural ...
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Background: Candida albicans is one of the most important opportunistic pathogens that suppress immunologic mechanisms of the host. It is speculated that structural and secretory proteins of C. albicans have immunomodulatory effects in cancer. Objective: To evaluate the effects of C. albicans structural and secreted proteins on intratumoral CD4/CD8 ratio as well as the survival rate in BALB/c tumor model. Methods: Structural and secretory proteins from C. albicans were isolated and examined for their effects on tumor growth and survival of adenocarcinoma bearing mice. Results: The results indicated that in mice treated with C. albicans structural protein, the survival rate significantly decreased compared with the control groups. Also, mice treated with secretory proteins showed a decrease in survival rate but it was not statistically significant (p>0.05). Investigating the frequency of tumor infiltrated CD4+ and CD8+ T lymphocytes indicated that the percentages of tumor infiltrated CD4+ T lymphocytes in response to structural and secreted proteins were higher compared to the control groups. Conclusion: Our study suggests that C. albicans structural and secreted proteins modulate intratumor T lymphocyte infiltration.
Ali Memarian; Mahmood Jeddi Tehrani; Parvaneh Vossough; Ramazan Ali Sharifian; Hodjatallah Rabbani; Fazel Shokri
Volume 4, Issue 3 , December 2007, , Pages 145-154
Abstract
Background: Wnt molecules play a key role in growth, proliferation and development of some embryonic and adult organs as well as hematopoietic stem cells. Wnt signaling pathways are aberrantly activated in many tumor types, including solid tumors and hematologic malignancies. Objective: To investigate ...
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Background: Wnt molecules play a key role in growth, proliferation and development of some embryonic and adult organs as well as hematopoietic stem cells. Wnt signaling pathways are aberrantly activated in many tumor types, including solid tumors and hematologic malignancies. Objective: To investigate the expression profile of a large number of Wnt genes in leukemic cells from Iranian patients with acute myeloblastic leukemia. Methods: RT-PCR method was used to determine the Wnt genes expression in bone marrow (BM) and/or peripheral blood (PB) samples from 16 patients with AML and PB samples of 36 normal subjects. Results: Among 14 Wnt molecules included in this study, Wnt-7A and Wnt-10A were significantly down-regulated (p = 0.002 and p < 0.0001, respectively) and Wnt-3 was significantly over-expressed (p < 0.02) in AML patients compared to normal subjects. No significant association was found between Wnt expression and FAB classification of the patients. Conclusion: Our results demonstrated for the first time aberrant expression of Wnt-7A, Wnt-10A and Wnt-3 genes in Iranian AML patients. This may be of relevance to the tumorigenesis process in this malignancy.
Bahram Kazemi; Negar Seyed; Mojgan Bandehpour; Zarrin Sharifnia; Parviz Pakzad
Volume 5, Issue 3 , September 2008, , Pages 148-155
Abstract
Background: Although a simple and direct method does not exist for the detection of chlamydial infections, there are situations in which reliable serological tests, with sensi-tivity related to a specific antigen, could be helpful. Objective: The aim of this study was to clone the first 1100 bp of the ...
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Background: Although a simple and direct method does not exist for the detection of chlamydial infections, there are situations in which reliable serological tests, with sensi-tivity related to a specific antigen, could be helpful. Objective: The aim of this study was to clone the first 1100 bp of the C. trachomatis outer membrane protein 2 (omp2) gene in order to prepare a recombinant protein for use in an ELISA system designed to recognize the anti- C. trachomatis antibody in patient sera. Methods: The PCR product of the chlamydial omp2 gene was cloned in pBluescript and its first 1100 bp was sub-cloned in the pQE-30 expression vector and induced by IPTG. The recombinant protein was purified by affinity chromatography and its purity was confirmed by SDS-PAGE, gel diffusion and western blot analyses. The purified protein was coated onto a polysty-rene microplate and tested by ELISA using patient serum. Results: We have cloned, over-expressed and purified biologically functional recombinant truncated Omp2 from C. trachomatis for use, as a species-specific recognition antigen, in an ELISA system. In this study we determined a cut-off value of 0.345 for this ELISA system using 55 negative sera and measured six positive sera at dilutions of 1:20-1:2560. Conclusion: As a species-specific recognition antigen, the over-expressed and purified recombinant truncated Omp2 from C. trachomatis performed well in an ELISA system.
Elfadil Abass; Abdelhafeiz Mahamoud; Durria Mansur; Mehdi Mohebali; Abdollah el Harith
Volume 8, Issue 3 , September 2011, , Pages 150-158
Abstract
Background: A β-mercaptoethnol (β-ME)-treated promastigote antigen of L. donovani was successfully employed in direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis (VL). Objective: The β-ME-treated antigen was further incorporated into an enzyme-linked immunosorbent ...
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Background: A β-mercaptoethnol (β-ME)-treated promastigote antigen of L. donovani was successfully employed in direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis (VL). Objective: The β-ME-treated antigen was further incorporated into an enzyme-linked immunosorbent assay set-up (β-ME ELISA) and evaluated for VL diagnosis against outcome of reference freeze-dried DAT (FD-DAT) and rK39 strip test (RKT) commercial kits. Methods: Two-hundred and ninety-two sera from patients with high VL suspicion of whom 105 had confirmed L. donovani infection were tested. Results: Relatively higher sensitivities of 93.3% (95% CI: 88.4- 98.2) and 92.4% (95% CI: 87.3-97.5) were determined for β-ME ELISA and FD-DAT as compared to 83.8% (95% CI: 76.7-90.8) for RKT. Of 73 VL sera that scored maximum absorbance values (>0.81) in β-ME ELISA, 66 (90.4%) tested at the highest agglutination titres (>1:51200) in FD-DAT as did 56 (76.7%) also at comparable reaction intensities (3 + colour intensity) in RKT. Compared with FD-DAT (94.7%, 95% CI: 91.5-97.9) or RKT (93.0%, 95% CI: 89.3-96.6), lower specificity was estimated for β-ME ELISA (90.4%, 95% CI: 86.1-94.6). Based both on positive and negative microscopy for L. donovani in organ aspirates of all VL suspects enrolled (292), significantly higher correlation (p<0.01, 0.919) was established between β-ME ELISA and FD-DAT than between β-ME ELISA and RKT (p<0.01, 0.824). Taking into calculation the combined estimates of sensitivity, specificity, positive and negative predictive values, higher agreement (94.8%) was determined between total performance of β-ME ELISA and FD-DAT than between that of β-ME ELISA and RKT (90.7%). Conclusion: Based on results and merits discussed, we recommend application of this β-ME ELISA both for diagnosis of VL at laboratory level and confirmation of results obtained with DAT or RKT in the field.
Afsoon Shariat; Mohammad Hossein Karimi; Talat Mokhtariazad; Seyed Mohammad Moazzeni; Bita Geramizadeh; Seyed Ali MalekHosseini; Ramin Yaghobi
Volume 11, Issue 3 , September 2014, , Pages 153-165
Abstract
Background: Dendritic cells (DCs) are potent antigen presenting cells for triggering of the immune reaction post transplantation. These cells are centrally involved in the initiation of T cell-dependent immune responses. Objective: To compare the level of DC maturation and function in liver transplant ...
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Background: Dendritic cells (DCs) are potent antigen presenting cells for triggering of the immune reaction post transplantation. These cells are centrally involved in the initiation of T cell-dependent immune responses. Objective: To compare the level of DC maturation and function in liver transplant recipients with healthy controls. Methods: In this study, twelve peripheral blood samples were selected from six liver transplant patients and six healthy controls. After the generation of DCs from monocytes, expression levels and mean fluorescent intensity (MFI) of several DC maturation markers were evaluated using flowcytometry. Secretion of IL-6, IL-12 and IL-23 proinflammatory cytokines was determined using ELISA. Gene expressions of TLR-2, TLR-4 and IL-23 were analyzed using real-time PCR. Results: DC expression markers including CD83 (p=0.007) and CD86 (p=0.02), as well as secretion of IL-6 (p=0.02) and IL-12 (p=0.007) by DCs were significantly increased in liver transplant patients compared with healthy controls. The MFI of CD86 (p=0.009) and HLA-DR (p= 0.005) expression on DCs was also higher in patients. The expression of TLR-2 transcripts in DCs of patients was higher than that of the controls (p=0.03). Conclusion: Based on these findings, increased frequency of DCs expressing CD83 and CD86, higher expression of CD86, HLA-DR, and TLR-2 as well as elevated secretion of proinflammatory cytokines in DCs of liver transplant recipient's point to the more mature phenotype and active function of DCs in patients compared with controls.
Fatemeh Hajighasemi; Ali Akbar Saboor-Yaraghi; Fazel Shokri
Volume 1, Issue 3 , December 2004, , Pages 154-161
Abstract
Background: The affinity of an antibody to its antigen is a crucial parameter in its biological activity and performance of an immunoassay such as ELISA. Affinity of most IgG specific MAbs are often determined by methods which require labeling of either antigen or antibody, and are sometimes difficult ...
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Background: The affinity of an antibody to its antigen is a crucial parameter in its biological activity and performance of an immunoassay such as ELISA. Affinity of most IgG specific MAbs are often determined by methods which require labeling of either antigen or antibody, and are sometimes difficult to control, do not always lead to the expected signal and often result in immunological modification of the molecules. Moreover, direct solid phase binding assays pose some problems such as diffusion effects and difficulties in reaching equilibrium due to heterogeneous binding and co-operativity. Objective: To employ a rapid and simple ELISA-based method for measuring affinity constants of two pan-h-IgG specific MAbs (3F2D8 and 5F19G11) established in our laboratory. Methods: The method is based on the effect of antibody affinity on the sigmoidal dose response curve. In this method, the binding of anti-human IgG (anti-h-IgG) MAbs with their corresponding antigen was measured using serial concentrations of both antigen and antibody. The amount of antibody bound to the antigen on the plate is represented as a sigmoidal curve of OD versus the logarithm of antibody concentration added to each well. Results: Based on the data obtained from this study, the affinity constants of 3F2D8 and 5F19G11 MAbs were 0.74 x 10 8 Mol –1 and 0.96 x 10 7 Mol –1, respectively. Conclusion: 3F2D8 MAb with reasonably high affinity is suggested as a candidate for quantitative measurement of IgG by ELISA, whereas 5F19G11 MAb could be considered as a suitable tool for immunoaffinity chromatography.
Mehri Ghafourian Boroujerdnia; Fatemeh Ghalambor Dezfuly; Nepton Emad Mosthophy; Rahim Chinipardaz
Volume 3, Issue 4 , December 2006, , Pages 157-163
Abstract
Background: Recent attention has focused on the expression of integrin molecules within the endometrium, and their relation to infertility. Objective: The present prospective study was undertaken to determine whether the endometrium of women with unexplained infertility differs in the expression of very ...
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Background: Recent attention has focused on the expression of integrin molecules within the endometrium, and their relation to infertility. Objective: The present prospective study was undertaken to determine whether the endometrium of women with unexplained infertility differs in the expression of very late activation antigens (VLA) from the endometrium of normal fertile women. Methods: Thirty samples of endometrial biopsies from hysterectomies with non-endometrial pathology and 28 endometrial samples by uterine curetting from infertile women in secretary phase at implantation time were collected, stained with three monoclonal antibodies against β1 integrin subunits including VLA-1 to VLA-3 by immunohistochemical technique and then assessed semi-quantitatively by microscope. Chi-Square test was used to compare the expression of VLA antigens on epithelial cells, stromal cells, lymphocytes and vessels within endometrial tissues between two groups. Results: The results showed that most VLA integrins were present in fertile and infertile endometrium tissues. There were similarities and differences in the expression of VLA molecules in different compartments. VLA-2, VLA-3 expression on endometrial compartments showed an unaltered pattern of staining during the putative window of implantation in either fertile or infertile women with no significant differences (Pvalue> 0.5). VLA-1 expression on endometrial compartments changed in fertile and unexplained infertile women, the differences were related to the presence or lack of the molecules on epithelial and stromal cells respectively. Conclusion: Differences may explain causes of unexplained infertility, and suggests that certain integrins may participate in the cascade of molecular events leading to successful implantation and early placental development which requires more investigations.
Alireza Rafiei; Mahoud Abedini; Seyed Hamzeh Hosseini; Zahra HosseiniKhah; Behrouz Bazrafshan; Mohsen Tehrani
Volume 9, Issue 3 , September 2012, , Pages 159-167
Abstract
Background: The pathogenesis of migraine involves immune-mediated mechanisms in the vascular endothelium. Toll like receptor 4 (TLR-4) is a signaling receptor of innate immunity which plays a role in various neuropathologies related to neuron inflammation. Objective: This case/control study is aimed ...
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Background: The pathogenesis of migraine involves immune-mediated mechanisms in the vascular endothelium. Toll like receptor 4 (TLR-4) is a signaling receptor of innate immunity which plays a role in various neuropathologies related to neuron inflammation. Objective: This case/control study is aimed to investigate whether TLR- 4 896A/G variation is related to migraine headaches in an Iranian population. Methods: A total of 170 migraine patients (130 females, mean age 33.24 ± 11 years) and 170 age, sex, and ethnicity matched healthy controls (118 females, mean age of 31 ± 10 years) were recruited. Genotyping was carried out using the tetra primer amplification refractory mutation system (ARMS)-PCR. Results: The frequency of G allele was higher in migraine patients than the controls (15% vs. 4.7%; p<0.0001). Interestingly, the distribution of heterozygous 896A/G genotype statistically differed between migraineurs and controls (25.3% vs. 8.2%, p=0.00002, OR 3.87, 95% CI; 2.02-7.4). Multivariate logistic regression analysis indicated that G allele in affected female migraineurs is an independent factor associated with increased risk of migraine (OR 3.2, 95% CI 1.23-8.24, p=0.01). Conclusion: Our results showed TLR-4 polymorphism as a genetic risk factor for migraine. However, further studies in different populations are required to elucidate the precise role of TLR-4 896A/G mutation in susceptibility to migraine.
Shole Daneshvar-ghahfarokhi; Amir Rahnama; Vahid Mohammadi-Shahrokhi
Abstract
Background: One of the inflammatory diseases of the respiratory system is asthma. Teucrium polium (TP) has anti-inflammatory and anti-allergic properties and its anti-asthmatic effects have not been investigated yet. RORγt is an inflammatory transcription factor for Th17 differentiation. By secreting ...
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Background: One of the inflammatory diseases of the respiratory system is asthma. Teucrium polium (TP) has anti-inflammatory and anti-allergic properties and its anti-asthmatic effects have not been investigated yet. RORγt is an inflammatory transcription factor for Th17 differentiation. By secreting IL-17, Th17 leads to neutrophilic inflammation in the lungs. As an anti-inflammatory cytokine, IL-10 reduces the dissemination of inflammatory elements in the airways.Objective: To evaluate the effect of TP extract in asthma treatment.Methods: Thirty female Balb/c mice were distributed into 5 groups (n=6) including the control, treated with ovalbumin (OVA), and OVA+ various doses of TP (50, 150, and 300 mg/kg). All groups except the control group were sensitized to OVA solution on days 0, 7, and 14 by subcutaneous injection. The challenge was performed on days 18 to 21 by the inhalation of 1% OVA and the treatment was done with TP extract in the treatment groups, half an hour before the challenge. On day 22, the serum and spleen samples were collected to determine IL-10 serum levels and RORγt gene expression, respectively.Results: In the treatment groups, the expression of RORγt significantly decreased when using OVA+ Tp extract (150 mg/kg and 300 mg/kg), and IL-10 serum levels significantly increased when using OVA+ TP extract (150 mg/kg) compared with the OVA group.Conclusion: It is possible that TP extract can be effective in improving asthma by reducing inflammation.
Hamid-Reza Zare; Mojtaba Habibagahi; Akbar Vahdati; Zahra Habibagahi
Volume 12, Issue 3 , September 2015, , Pages 166-175
Abstract
Background: Patients with rheumatoid arthritis (RA) suffer from wide ranges of autoimmune reactions in joints. The mechanism of which is generally unknown and maybe associated with Treg deregulation. Objective: To compare the frequency of nTregs in peripheral blood of patients with active rheumatoid ...
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Background: Patients with rheumatoid arthritis (RA) suffer from wide ranges of autoimmune reactions in joints. The mechanism of which is generally unknown and maybe associated with Treg deregulation. Objective: To compare the frequency of nTregs in peripheral blood of patients with active rheumatoid disease with healthy individuals. Methods: Twenty five newly diagnosed patients with active RA disease were selected based on the clinical and laboratory criteria before starting their therapies. Treg cells in peripheral blood samples were enumerated by immune staining and flowcytometry analysis. Results: Clinical and laboratory results were in favor of active disease in all the studied patients although they showed variations in Disease Activity Score-28 (DAS-28). Compared to the healthy controls, RA patients had significantly lower frequency of CD4+ CD25hi or CD4+ CD25+ FoxP3+ natural regulatory T cells. In spite of that, there were no significant differences between patients and healthy controls in respect to the CD4/CD8 ratio. Interestingly, more CD4+ CD25- FoxP3+ cells were found in peripheral blood of patients. The frequencies of the Tregs did not show strong associations with the DAS-28. Conclusion: We showed lower abundance of nTregs in peripheral blood of RA patients which highlights the significance of these cells in RA.
Hamideh Mesali; Abolghasem Ajami; Hadi Hussein-Nattaj; Alireza Rafiei; Zeinab Rajabian; Hossein Asgarian-Omran; Vahid Hosseini; Tarang Taghvaei; Mohsen Tehrani
Volume 13, Issue 3 , September 2016, , Pages 167-177
Abstract
Background: Regulatory T Cells (Tregs) and Myeloid-Derived Suppressor Cells (MDSCs) are two main regulatory cells modulating the immune responses in inflammation and cancer. Objective: To investigate and compare Tregs and MDSCs in peptic ulcer and gastric cancer. Methods: Patients with dyspepsia were ...
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Background: Regulatory T Cells (Tregs) and Myeloid-Derived Suppressor Cells (MDSCs) are two main regulatory cells modulating the immune responses in inflammation and cancer. Objective: To investigate and compare Tregs and MDSCs in peptic ulcer and gastric cancer. Methods: Patients with dyspepsia were selected and divided into three groups of non-ulcer dyspepsia (NUD, n=22), peptic ulcer disease (PUD, n=25), and gastric cancer (GC, n=27) according to their endoscopic and histopathological examinations. Helicobacter pylori infection was diagnosed by rapid urease test and histopathology. The number of peripheral blood CD4+CD25+FoxP3+Tregs and CD14+HLA-DR- MDSCs were determined in all patients, by flow cytometry. The number of FoxP3+ regulatory T cells was also determined by immunohistochemistry (IHC). Results: The percentage of peripheral blood Treg cells in both PUD )0.81 ± 0.39, p<0.001) and GC groups )0.98 ± 0.65, p<0.001) were significantly higher than in NUD group (0.46 ± 0.10). These results were also confirmed by IHC. A significantly higher percentage of MDSCs in patients with PUD )0.73 ± 0.19, p<0.001) and GC )0.73 ± 0.16, p<0.001) was also observed when compared to NUD group )0.46 ± 0.16). There was no difference in the percentages of these two cell types between the PUD and GC groups. The percentages of Tregs and MDSCs in patients with PUD and GC were not significantly correlated. Conclusions: Both Tregs and MDSCs showed higher frequencies in PUD and GC. These results suggest that immune-modulation by the Tregs and MDSCs may play a role in the pathogenesis of PUD and GC.
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.
Fatemeh Nasri; Mehrnoosh Doroudchi; Bahia Namavar Jahromi; Behrouz Gharesi-Fard
Abstract
Background: Polycystic ovary syndrome (PCOS) is considered as the most common cause of female infertility that affects 4-10% of women in the reproductive age. Previous studies have shown the role of a balanced immune response in a successful pregnancy and fertility. Objective: To investigate the T helper ...
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Background: Polycystic ovary syndrome (PCOS) is considered as the most common cause of female infertility that affects 4-10% of women in the reproductive age. Previous studies have shown the role of a balanced immune response in a successful pregnancy and fertility. Objective: To investigate the T helper cells type 1 (Th1) /Th2/Th17/Treg paradigms in peripheral blood of infertile PCOS compared with normal fertile women. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated at the late follicular phase from 10 PCOS and 10 fertile women. PBMCs were stimulated with PMA and ionomycin in the presence of Berefeldin A as Golgi stop agent to detect intracellular cytokine production (IFN-γ, IL-17, and IL-4) from CD3+CD4+T cells population indicating T helper (Th) cells subsets by flowcytometry. Moreover, regulatory T cells were enumerated using CD25 and Foxp3 markers. Results: In this study, we report that the frequency of Th1 cells was increased compared to Th2 cells in infertile PCOS when considering Th1/Th2 ratio (P=0.05). Analysis of Th17/Th2 ratio showed a significant difference with a bias toward Th17 dominancy in PCOS (P=0.02). The proportion of CD4+CD25+Foxp3+ regulatory T cells was significantly lower in PCOS patients than that of healthy fertile women (P=0.02). Conclusion: In summary, Th1 and Th17 bias and reduction of Treg and Th2 cells as regulators of immune responses might be involved in the pathogenesis of PCOS. These results are suggestive of an altered immune response to inflammatory status in PCOS patients, likely causing some complications such as infertility in these patients.