yunlong zhuang; xixi li; huicong xu; hui ye; di sun; xiangzhng liu; Guijie Ren
Abstract
Background: Entecavir (ETV) is commonly used to treat chronic hepatitis B (CHB) in China. However, certain percentages of e-Antigen (HBeAg) positive CHB patients do not respond to ETV therapy. Objective: To investigate whether the killer immunoglobulin-like receptor (KIR) genes were associated with seroconversion ...
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Background: Entecavir (ETV) is commonly used to treat chronic hepatitis B (CHB) in China. However, certain percentages of e-Antigen (HBeAg) positive CHB patients do not respond to ETV therapy. Objective: To investigate whether the killer immunoglobulin-like receptor (KIR) genes were associated with seroconversion in HBeAg positive CHB responder patients treated with ETV. Methods: Polymerase chain reaction with sequence-specific primers (PCR-SSP) method was performed to genotype KIR genes in 200 healthy controls and 198 HBeAg-positive CHB patients which 59 were defined as the complete response group (CRG) to the treatment with ETV and 139 were defined as null or partial response group (NPRG). Results: The frequencies of KIR2DS2 and KIR2DS3 were significantly higher (P=0.030, OR=1.57,95%CI=2.36-1.05 and P=0.018, OR=1.773,95%CI=2.77-1.13, respectively), while, the frequencies of KIR2DL3, KIR2DS1 and KIR3DS1 were significantly lower (P=0.038, OR=0.525, 95%CI=0.96-0.29,and P=0.031, OR=0.640, 95%CI =0.95-0.43, and P=0.035, OR=0.641, 95%CI =0.96-0.43, respectively) in HBeAg-positive CHB patients than those in healthy controls. The frequency of KIR2DS3 gene was significantly higher in NPRG than that in CRG (P=0.018, OR=0.402, 95%CI=0.83-0.20). The frequencies of KIR2DL3 and KIR3DS1 genes were significantly higher in CRG than those in NPRG (P=0.019, OR=3.625, 95%CI=10.83-1.21 and P=0.041, OR=1.949, 95%CI=3.65-1.04, respectively). Conclusion: Patients with KIR2DS3 might have negative responses to anti-HBV therapy with ETV and patients with KIR2DL3 and KIR3DS1 might have advantage in the therapy with ETV.
Faramarz Dobakhti; Soheila Ajdari; Mohammad Taghikhani; Shahnaz Rafiei; Khosrow Bayati; Mortez Rafiee-Tehrani
Volume 3, Issue 3 , September 2006, , Pages 114-120
Abstract
Background: Different methods have been used for BCG vaccination. Alginate microspheres are useful in delivery of vaccines to the gastrointestinal tract by oral route. Objective: To compare the immune response following oral microencapsulated and subcutaneous (SC) route administration of BCG vaccine ...
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Background: Different methods have been used for BCG vaccination. Alginate microspheres are useful in delivery of vaccines to the gastrointestinal tract by oral route. Objective: To compare the immune response following oral microencapsulated and subcutaneous (SC) route administration of BCG vaccine in BALB/c mice. Methods: Alginate microspheres were produced by an internal emulsification method within olive oil. Four groups of mice were studied, including two groups receiving oral gavages of microencapsulated and free BCG, one receiving SC injection of BCG, and a control group. T cell proliferation, specific anti-BCG total IgG, and IgG subclasses (IgG1 and IgG2a) were compared between groups 5 and 12 weeks after vaccination. Results: The best result was achieved using oral microencapsulated form in comparison with oral BCG alone. Conclusion: Delivery of oral BCG with alginate microspheres is an effective way to induce immune response in BALB/c mice.
Hadi Hossein-Nataj; Mohsen Masjedi; Mohammad Hassan Emami; Mojgan Mokhtari; Fereshteh Alsahebfosoul
Abstract
Background: Increased number of intestinal intraepithelial lymphocytes (IELs) is a key histological finding in the diagnosis of celiac disease (CD); however, the number of IELs in celiac patients and healthy subjects may vary from one region to another. Additionally, there are some seronegative celiac ...
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Background: Increased number of intestinal intraepithelial lymphocytes (IELs) is a key histological finding in the diagnosis of celiac disease (CD); however, the number of IELs in celiac patients and healthy subjects may vary from one region to another. Additionally, there are some seronegative celiac patients with a borderline histology. Objective: To determine the number of the CD3+ and CD8+ IELs T-cells in the celiac patients and healthy subjects (controls) in Isfahan. Methods: The duodenal biopsies were obtained from the celiac patients (n=15) and the controls (n=19). The total number of IELs/100 epithelial cells (ECs) were counted using the hematoxylin-eosin (H&E) staining method, and that of CD3+ and CD8+ IELs/100 ECs were counted using the immunohistochemistry (IHC) staining method. Results: This study defined the upper normal limit for each variable as mean + 2SD. Accordingly, the upper normal limits of the total IELs, CD3+ IELs, and CD8+ IELs/100 ECs were calculated as 37 (95% confidence intervals, CI: 33–41), 22 (95% CI: 19–25) and 12 (95% CI: 10–14), respectively. In 3 clinically CD diagnoses, the total IELs counts/100 ECs were below the upper normal limit, and the histopathological and serologic assays were negative. Nevertheless, the CD8+ IELs T-cells counts/100 ECs showed borderline values. Interestingly, these patients responded to a gluten-free diet (GFD). Conclusions: The study findings suggest that in the clinically diagnosed celiac disease, IELs count/100 ECs below the upper normal limit as well as negative histopathological and serologic assays and the cell density counts of the CD8+ IELs T-cells/100 ECs could be a useful parameter for CD diagnosis and make a decision to put them on a GFD.
Mohammad Hadi Fakoor; Parviz Owlia; Seyed Latif Mousavi gargari; Azar Sabokbar
Abstract
Background: Pseudomonas aeruginosa is considered as the most severe cause of infections in burn patients and pneumonia infections. Objective: To study the protective effects of recombinant protein vaccine harboring the PcrV of P. aeruginosa in the mouse model of burn and respiratory infections. Methods: ...
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Background: Pseudomonas aeruginosa is considered as the most severe cause of infections in burn patients and pneumonia infections. Objective: To study the protective effects of recombinant protein vaccine harboring the PcrV of P. aeruginosa in the mouse model of burn and respiratory infections. Methods: Recombinant protein vaccine harboring the PcrV was expressed in the E. coli BL-21 strain. Mice were immunized with the purified recombinant protein, and the antibody titer was measured in the sera obtained from the immunized mice. Immunized and control mice werechallenged by active and passive immunization. The microbial counts in the lung, skin, liver, spleen, and kidney were compared with the control mice. Results: Bioinformatics analysis indicated that the PcrV protein was conserved in 1552 clinical and environmental isolates. Also, the isoelectric point (pI), molecular weight, and Grand Average of Hydropathy (GRAVY) score were analyzed. Mice were injected with recombinant protein, and serum from immunized mice reacted strongly with recombinant antigen at a dilution of 1:64000. The survival rate of mice infected with 5xLD50 of the P. aeruginosa increased significantly up to 75% in the standard strains (PAO1 and PAK), and the number of bacteria, especially in the internal organs (kidney, spleen, and liver) significantly reduced compared to the mice immunized with placebo. Conclusions: Our results demonstrated that the PcrV protein could be an effective candidate vaccine for the generation of antibody response against P. aeruginosa infection.
Volume 6, Issue 3 , September 2009, , Pages 130-140
Mohammad Hasan Sheikhha; Mehdi Kalantar; Khalid Tobai; John A. Liu Yin
Volume 2, Issue 3 , September 2005, , Pages 141-151
Abstract
Background: The glutathione S-transferase (GST) family of metabolising enzymes plays an important role in the detoxification of mutagens and carcinogens. The expression of many of these cancer susceptibility enzymes is genetically polymorphic. An increased frequency of GST-null genotypes has been associated ...
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Background: The glutathione S-transferase (GST) family of metabolising enzymes plays an important role in the detoxification of mutagens and carcinogens. The expression of many of these cancer susceptibility enzymes is genetically polymorphic. An increased frequency of GST-null genotypes has been associated with several malignancies. Objective: To investigate the rate of GSTT1 and GSTM1 null genotypes in AML patients and to determine its importance in prognosis of the disease. Methods: DNA was extracted by phenol/chloroform method from peripheral blood or bone marrow of 180 white Caucasian patients. A multiplex PCR method was used simultaneously to amplify regions of GSTM1, GSTT1, and b-globin genes in genomic DNA. The survival curves were analyzed by the Kaplan-Meier method and compared by the log-rank test (Mantel-Cox) using the SPSS software program. Results: Of the total of 180 patients, 23 cases (12.8%) showed null genotypes in both genes, while in 52 patients (28.9%) both genes were wild-types. GSTM1 null-GSTT1 wild-type was detected in 91 patients (50.6%) and GSTM1 wild-type-GSTT1 null genotype was detected in 14 patients (7.8%). These rates are within the upper limit of the rates detected in the normal European population. There was no significant difference in the overall survival and in disease free survival between different groups. Conclusion: These observations suggest that the inherited absence of the GSTT1 and GSTM1 carcinogen detoxification pathway may be related to carcinogenesis but it is not an important determinant of prognosis in AML.
Elham Ashouri; Mohammad Hossein Dabbaghmanesh; Amirhossein Hadaegh; Soodeh Rowhanirad; Marizeh Bakhshayeshkaram; Gholamhossein Ranjbar Omrani
Volume 10, Issue 3 , September 2013, , Pages 150-157
Abstract
Background: Killer cell immunoglobulin-like receptors (KIR) are expressed on NK cells and a subset of T cells. The variable KIR receptors along with their ligands, HLA class I, influence risk for autoimmune and malignant diseases. Objective: To investigate the KIR gene profiles in relation to susceptibility ...
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Background: Killer cell immunoglobulin-like receptors (KIR) are expressed on NK cells and a subset of T cells. The variable KIR receptors along with their ligands, HLA class I, influence risk for autoimmune and malignant diseases. Objective: To investigate the KIR gene profiles in relation to susceptibility to Graves’ disease in patients with ophthalmopathy. Methods: KIR genes profiles were analyzed in 90 patients presenting Graves’ disease with ophthalmopathy representing upper eyelid retraction, swelling, redness, conjunctivitis, and bulging eyes and were compared with the KIR gene profiles of 112 healthy controls. The presence and absence of 11 variable KIR genes were characterized using a gene-specific PCR typing system. Results: There was no significant difference in the distribution of KIR gene profiles between patients and controls. Conclusion: Our data show that none of the KIR genotypes contribute in susceptibility to Graves’ disease; although the role of HLA ligand remains to be characterized.
Babacar Mbengue; Birahim Niang; Bacary Diatta; Adama Tali; Olivier Garraud; Ronald Perraut; Alioune Dieye
Volume 7, Issue 3 , September 2010, , Pages 150-161
Abstract
Background: Cerebral malaria (CM) is one of the major causes of death in African populations infected with Plasmodium falciparum. Only 1% of infected subjects develop CM. The reasons for these differences are not fully understood, but it is likely that the host humoral response against blood-stage antigens ...
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Background: Cerebral malaria (CM) is one of the major causes of death in African populations infected with Plasmodium falciparum. Only 1% of infected subjects develop CM. The reasons for these differences are not fully understood, but it is likely that the host humoral response against blood-stage antigens plays a role in protection from malaria, although the precise targets and mechanisms mediating immunity remain unclear. Objective: The purpose of this study was to distinguish between defined P. falciparum- specific Ab response patterns in patients presenting with mild malaria (MM) vs. CM. Methods: We used a panel of P. falciparum conserved antigens including crude blood-stage extracts schizont, merozoite and parasitised erythrocyte membranes and MSP-1p19, PfEB200, R23 and GST-5 recombinant antigens in a retrospective casecontrol study of symptomatic adults, one group presenting confirmed CM without fatal outcome and another group with MM. We further matched P. falciparum-specific Ab responses with those from individuals living in an endemic setting known to have protective immunity and considered them as “immune control” subjects (IC). Total IgG, IgM and IgG subclass Ab responses were determined using ELISA method. Results: Substantial Ab responses were found in symptomatic patients, significantly lower than the “immune control” subjects, and with a limited quantitative difference between MM versus CM. Interestingly, asynchronous IgM response was evidenced in CM contrary to MM. Conclusion: Our results suggest that the contribution of an efficient IgG response against parasite multiplication is of importance in the evolution towards CM manifestation without fatal outcome and deserves further analysis for vaccine candidates.
Chaohui Zhu; Min Fan; Jianhua Zhu; Limin Cao; Xinyu Duan; Kai Wu
Abstract
Background: Vitamin D has anti-inflammatory efficacy against ulcerative colitis (UC), however, the mechanism is yet little understood. Objective: To investigate the immunomodulatory effects of vitamin D against the UC, and to explore the potential downstream mechanisms. Materials and methods: Serum vitamin ...
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Background: Vitamin D has anti-inflammatory efficacy against ulcerative colitis (UC), however, the mechanism is yet little understood. Objective: To investigate the immunomodulatory effects of vitamin D against the UC, and to explore the potential downstream mechanisms. Materials and methods: Serum vitamin D, Interferon-γ (IFN-γ) and Interleukin (IL)-17 levels of the patients with UC were quantified using enzyme-linked immunosorbent assay (ELISA). Long non-coding RNAs (lncRNAs) levels were determined by using quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Peripheral blood mononuclear cells (PBMCs) were collected from healthy control subjects, stimulated with CD4+ T lymphocytes or helper T cells 17(Th17) differentiation conditions, and then exposed to calcitriol (vitamin D active form) or certain lentiviral treatment, followed by subsequent molecular level testing. For in vivo assay, mice were given 3% dextran sulfate sodium (DSS) to induce colitis. Results: Compared with the control group, vitamin D levels in the UCs were statistically lower, and there was a negative correlation between IL-17 and vitamin D in the UCs. The lncRNA OIP5-AS1 could decrease under calcitriol treatment in both CD4+ T cells and Th17 differentiation. The lncRNA OIP5-AS1 was a microRNA (miR)-26a-5p sponge and therefore modulated the Th17 cells and IL-6 expression. The lncRNA OIP5-AS1/miR-26a-5p/IL-6 axis mediated the regulation of calcitriol-induced Th17 differentiation. Calcitriol had therapeutic effects on the UC mouse models by regulating the lncRNA OIP5-AS1 related pathway. Conclusion: Vitamin D might have anti-inflammatory potential in the treatment of the UC.
Eskandar Kamali-Sarvestani; Hadi Farsiani; Michel Shamoon Pour; Abdulah Bazargani; Kamran Lankarani; Ali-Reza Taghavi; Mehdi Saberifiroozi
Volume 4, Issue 3 , December 2007, , Pages 155-160
Abstract
Background: Polymorphisms in the immune related genes are important in the clinical outcome of Helicobacter pylori infection. Myeloperoxidase -463 G/A polymorphism has been shown to reduce enzyme expression and activity. Objective: the aim of the present study is to investigate the association of myeloperoxidase ...
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Background: Polymorphisms in the immune related genes are important in the clinical outcome of Helicobacter pylori infection. Myeloperoxidase -463 G/A polymorphism has been shown to reduce enzyme expression and activity. Objective: the aim of the present study is to investigate the association of myeloperoxidase G-463A polymor-phism with clinical outcome of Helicobacter pylori infection. Methods: two hundred and eighty five patients with positive culture of Helicobacter pylori from their gastric biopsies are included in this study. Human leukocyte DNA was extracted using salting out method and myeloperoxidase G-463A polymorphism was investigated by PCR-RFLP. All clinicopathological data were collected from individual records. Results: When the patients were categorized according to the high (GG) and low + intermediate (AG+AA) genotypes of myeloperoxidase producers, there was a significant association between myeloperoxidase G-463A genotypes and clinical outcome of Helicobacter py-lori infection (p=0.006). In search for combined effect of cagA status and myeloperoxi-dase genotypes on clinical presentations, only in cagA- Helicobacter pylori infected pa-tients a significant association between myeloperoxidase genotypes and clinical out-come was found (p=0.0001). Also this association was found only in patients infected with vacA s1m1 genotype (p=0.008). Conclusions: Our findings suggest that the mye-loperoxidase G-463A polymorphism is a host genetic factor which determines the clini-cal outcome of Helicobacter pylori infection. Moreover, the combination of host and bacterial genetics could provide a better understanding of clinical outcome after infec-tion with Helicobacter pylori.
Mojtaba Sankian; Forough Glosaze Shirazi; Mahid Arafi; Malihe Moghadam; Abdolreza Varasteh
Volume 5, Issue 3 , September 2008, , Pages 156-162
Abstract
Background: Allergy to Saffron (Crocus sativus) pollen has been described in people involved in processing of saffron flower stamens. Profilins have been identified as a pan-allergen in different plant pollens and foods. This molecule is an actin-binding pro-tein with a molecular weight of 12-16 kDa ...
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Background: Allergy to Saffron (Crocus sativus) pollen has been described in people involved in processing of saffron flower stamens. Profilins have been identified as a pan-allergen in different plant pollens and foods. This molecule is an actin-binding pro-tein with a molecular weight of 12-16 kDa found in eukaryotic species. Objective: The aim of this study was to generate monoclonal antibody against Cro s 2 in order to char-acterize this major allergen of saffron pollen. Methods: BALB/c mice were immunized to obtain adequate humoral response. Splenocytes were prepared from the immunized animals, mixed with the P3-X63-Ag8.653 myeloma cells and fused by means of PEG 1500. After two weeks of culturing in HAT-containing media, the supernatant from those wells growing hybridomas were screened by ELISA using plates coated with Cro s 2. Cells from positive wells were cloned at least 3 times by limiting dilution. Specific-ity and cross-reactivity of the mAbs were determined by Western blot analysis and sandwich ELISA. Results: Two stable hybridoma clones secreting mAbs against Cro s 2 were obtained and expanded. The anti-Cro s 2 mAbs were also found to cross-react with other plant profilins. Isotype of this mAb was identified as μ heavy chain and k light chain. Conclusion: The anti-Cro s 2 mAb could be a useful tool for characteriza-tion and standardization of many pollen and fruit-derived profilins.
Zahra Amirghofran; Saeed Malek-Hosseini; Hossein Golmoghaddam; Fathollah Kalantar; Mehdi Shabani
Volume 8, Issue 3 , September 2011, , Pages 159-169
Abstract
Background: A number of medicinal plants have been used to treat various immunological diseases. Nitric oxide (NO) has an important regulatory role in the various types of inflammatory processes. Objective: To investigate the NO modulatory activity of the extracts of several medicinal plants native to ...
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Background: A number of medicinal plants have been used to treat various immunological diseases. Nitric oxide (NO) has an important regulatory role in the various types of inflammatory processes. Objective: To investigate the NO modulatory activity of the extracts of several medicinal plants native to Iran including Dracocephalum kotschyi, Linum persicum, Dionysia termeana, Salvia mirzayanii, Ferulago angulata and Euphorbia cheiradenia. Methods: The methanolic extracts of the plants were prepared and examined for their effects on the NO production by lipopolysaccharide-stimulated mouse macrophages. The level of TNF-α and IL-1β proinflammatory cytokines in the macrophage culture were detected using enzyme-linked immunosorbent assay. Results: All the extracts at concentration of 50 μg/ml demonstrated a significant decrease in NO production (p<0.001) after a 24-hour treatment. This inhibitory effect was also seen after 48 hours. Among the extracts, L. persicum was the strongest extract in reducing the NO production at 1 μg/ml after both 24 and 48-hours (nearly 100% inhibition, p<0.001). S. mirzayanii extract with 66.2 ± 8% inhibition at 50 μg/ml, showed the mildest effects in 48 hour culture. In cytokine release determination, the extract of L. persicum significantly inhibited both TNF-α and IL-1β cytokines production by stimulated macrophages (p<0.001). D. kotschyi, D. termeana and F. angulata decreased secretion of IL-1β from the cells. Conclusion: These results indicate the presence of anti-inflammatory and macrophage inhibitory substances in these plants.
Behrouz Nikbin; Nader Tajik; Ali Saraji; Gholam Reza Pourmand; Fatemeh Talebian; Abdurasul Mehrsai; Ali Akbar Amirzargar
Volume 1, Issue 3 , December 2004, , Pages 162-168
Abstract
Background: The Presence of donor leukocytes in recipients of organ allograft has been shown even several years after transplantation. However, it remains unclear whether this donor cell microchimerism plays an effective role in allograft acceptance or is simply a consequence of immunosuppressive conditions ...
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Background: The Presence of donor leukocytes in recipients of organ allograft has been shown even several years after transplantation. However, it remains unclear whether this donor cell microchimerism plays an effective role in allograft acceptance or is simply a consequence of immunosuppressive conditions in recipients. Objective: To study microchimerism in a group of kidney transplant recipients. Methods: In this study, the Peripheral Blood Microchimerism (PBM) after renal transplantation was retrospectively evaluated in 32 male-to-female recipients of living (unrelated) and cadaveric donor renal transplants. Using a Nested Polymerase Chain Reaction (Nested-PCR) amplification specific for SRY region of the Y chromosome, microchimerism was detected with a sensitivity of 1:1000000. Recipients were classified and compared according to the presence of PBM, acute and chronic rejection episodes, type of allotransplant, recipient and donor age at transplantation, previous male labor or blood transfusion, allograft function (serum creatinine level), and body mass index. Results: Among 32 recipients, 7 (21.9) were positive for PBM in multiple testing at different post-transplantation times. All microchimeric recipients had received kidney from living-unrelated donors. No significant difference was observed with regard to other parameters mentioned above. In addition, acute rejection rate in the microchimeric group was 3 (42%) versus 4 (16%) in the nonmicrochimeric recipients (not significant). Conclusion: Our results demonstrate better establishment of microchimerism after living donor kidney transplantation. However, concerning the true effect of microchimerism after renal transplantation doubt still persists; and it seems that microchimerism alone has no major protective role in renal allograft survival.
Hojjatollah Shokri; Farzad Asadi; Ali Reza Bahonar; Ali Reza Khosravi
Volume 3, Issue 4 , December 2006, , Pages 164-168
Abstract
Background: Herbal medicines have been used since ancient times for treatment of a range of diseases and have represented stimulatory effects on the function of innate immunity. Objective: To evaluate the effects of Zataria multiflora (Z. multiflora) on the function of innate immunity including phagocytic ...
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Background: Herbal medicines have been used since ancient times for treatment of a range of diseases and have represented stimulatory effects on the function of innate immunity. Objective: To evaluate the effects of Zataria multiflora (Z. multiflora) on the function of innate immunity including phagocytic activity and TNF-α secretion in animal model. Methods: Eight BALB/c mice were divided into two equal groups. In group A, Z. multiflora essence was injected intraperitoneally to the mice, in group B, distilled water was injected. Blood was obtained from 4 mice in each group, 4 and 7 days following injection. The amounts of phagocytosis (respiratory burst) and TNF-α secretion were assessed by chemiluminescence and ELISA method, respectively. Results: Significant increase in phagocytosis and TNF-α secretion was observed in group A compared with the control group at days 4 and 7. Conclusion: Z. multiflora essence can remarkably stimulate innate immunity function and it may be used to immunize individuals alone or in combination with other immunostimulatory agents.
Yang Li; Lu Zhang; Shouyu Wang; Peng Shi; Wei Qu
Volume 11, Issue 3 , September 2014, , Pages 166-176
Abstract
Background: Fusion of dendritic cells (DCs) with melanoma cells could reinforce the antigenicity of tumors as a strategy for the treatment of malignant melanoma. However, the insufficient quantity of DCs and the low fusion efficiency limits the development of such approach. Objective: To define the ...
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Background: Fusion of dendritic cells (DCs) with melanoma cells could reinforce the antigenicity of tumors as a strategy for the treatment of malignant melanoma. However, the insufficient quantity of DCs and the low fusion efficiency limits the development of such approach. Objective: To define the dosage of the stimulating factors as well as the induction condition for the optimal DCs preparation and cell fusion. Methods: DCs were generated from murine bone marrow cells, and cultured with four different concentrations of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). DCs were confirmed to be mature by detecting the expression of MHC-II, CD11c, CD80, and CD83 by flowcytometry. DCs-melanoma fusion cells were generated using polyethylene glycols (PEG) with different molecular weights and the fusion efficiency was detected by fluorescence-activated cell sorter (FACS). Results: The largest quantity of DCs was found when cells were cultured with 1000 U/ ml of GM-CSF and 500 U/ml of IL-4 (1.69 ± 0.04 ×10 6 ml-1, p<0.001 when compared with the other three groups). The expression levels of MHC-II and CD83 on day 7 after incubation were significantly lower than those on day 3 (MHC-II: p<0.001; CD83: p<0.001). The efficiency of cell fusion under induction of PEG-3000 was significantly higher than that of PEG-4000 (15.4 ± 0.56% vs. 11.1 ± 0.45%, p<0.001). Conclusions: The largest quantity for mature DCs was stimulated with 1000 U/ml of GM-CSF and 500 U/ml of IL-4 and the highest fusion efficiency was under induction of PEG-3000.
Jianrong Niu; Hui Zhou; Rong Tian; Xudong Wang
Abstract
Background: Molecular markers are involved in atopic dermatitis (AD) pathogenesis. The estrogen receptor (ESR)-1 gene, encoding ERα, is reported to express aberrantly in AD patients.Objective: To detect the biological functions of ESR1 in 2,4 dinitrochlorobenzene (DNCB)-treated mice.Methods: The ...
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Background: Molecular markers are involved in atopic dermatitis (AD) pathogenesis. The estrogen receptor (ESR)-1 gene, encoding ERα, is reported to express aberrantly in AD patients.Objective: To detect the biological functions of ESR1 in 2,4 dinitrochlorobenzene (DNCB)-treated mice.Methods: The DNCB-treated mice received a topical application of emulsion containing the 1,3-bis(4 hydroxyphenyl)-4-methyl-5-[4-(2-piperidinyl ethoxy) phenol]-1H-pyrazole dihydrochloride (MPP; an ESR1-selective antagonist) to dorsal skins and ears. Then the dermatitis scores, histopathological changes, and cytokine levels were evaluated.Results: MPP specifically downregulated ESR1 expression in DNCB-applied mice. Functionally, application of MPP abolished the DNCB-induced promotion in dermatitis score. Additionally, MPP administration protected against DNCB-induced dermatitis severity, suppressed mast cell infiltration and reduced production of immunoglobulin E (IgE) and thymus and activation-regulated chemokine (TARC). Moreover, MPP treatment inhibited DNCB- induced production of Th2 cytokines and infiltration of CD4+ T cells.Conclusion: ESR1 facilitates Th2-immune response and enhances Th2 cytokines in AD mice.
Mohammad Hashem Soltani; Tahereh Kalantari; Mohammad Hossein Karimi; Nasrollah Erfani; Eskandar Kamali Sarvestani
Volume 9, Issue 3 , September 2012, , Pages 168-174
Abstract
Background: T helper 1 and T helper 17 cells play important roles in immunity against foreign invaders. Differentiation of these Th subsets is affected by state of maturation and cytokines that are produced by dendritic cells (DCs). Curdlan is a linear (1→3)-β- glucan and has shown activity ...
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Background: T helper 1 and T helper 17 cells play important roles in immunity against foreign invaders. Differentiation of these Th subsets is affected by state of maturation and cytokines that are produced by dendritic cells (DCs). Curdlan is a linear (1→3)-β- glucan and has shown activity against tumors and infectious agents. Objective: This study aims to investigate whether curdlan plays its role through affecting the maturation and cytokine production by DCs. Methods: DCs were isolated from the spleen of BALB/c mice by MACS method. After an overnight culture of DCs in the presence of curdlan, the expression levels of CD40, CD86, and MHC-II molecules were determined by flow cytometry. The production of cytokines involved in Th1 and Th17 cell differentiation (IL-12 and IL-6, respectively) was also evaluated by ELISA. Lipopolysaccharide (LPS) treated and untreated cells were considered as positive and negative controls, respectively. Results: The results of this study did not show a significant difference in the levels of surface expression of CD40 (p=0.82), CD86 (p=0.79), and MHC class II (p=0.84) molecules upon exposure to curdlan. However, LPS increased the intensity of CD40 expression on dendritic cells (p=0.04). In addition, it was revealed that curdlan-exposed DCs are not able to produce a significant amount of IL-6 and IL-12 cytokines. Conversely, LPS-treated DCs were able to make a significant amount of IL-12 (p=0.005). Conclusion: The results of the present study suggest that curdlan has no effect on Th1 or Th17 differentiation while LPS may induce Th1 deviation by induction of CD40 expression and IL-12 production.
Afshin Namdar; Hamid Reza Mirzaei; Farhad Jadidi-Niaragh; Mahboubeh Ashourpour; Maryam Ajami; Jamshid Hadjati; Abbas Rezaei
Volume 12, Issue 3 , September 2015, , Pages 176-187
Abstract
Background: Melanoma progression and metastasis is suggested to be mediated by increased accumulation of myeloid derived suppressor cells. Various chemotherapeutic drugs such as 5-Fluorouracil in single low concentration have the capacity, at least in part, to reverse tumor progression by reducing myeloid ...
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Background: Melanoma progression and metastasis is suggested to be mediated by increased accumulation of myeloid derived suppressor cells. Various chemotherapeutic drugs such as 5-Fluorouracil in single low concentration have the capacity, at least in part, to reverse tumor progression by reducing myeloid derived suppressor cellsmediated immunosuppression. Objective: To assess whether multiple low doses of 5- fluorouracil could repress myeloid derived suppressor cells in low frequency and, in turn, could enhance anti-tumor responses and promote a more prolonged survival in a murine melanoma model. Methods: Fifty milligram per kilogram body weight dose of 5-Flourouracil was administered intraperitoneally 4 times with 3-day intervals to C57BL/6 mice after B16 melanoma tumor models were established. The frequency and suppressive functions of myeloid derived suppressor cells and induction of anti-tumor CD8+ T cells as well as tumor growth and survival were evaluated in drug treated and untreated mice. Results: Our results demonstrated that this therapeutic strategy increases the overall mice survival (p≤0.01) and induces melanoma-specific CD8+T cell immunity (p≤0.01) by reducing the frequency of myeloid derived suppressor cells (p≤0.01) as well as their immune suppressive functions (p≤0.05). Conclusion: Altogether, our data suggest that 5-fluorouracil in multiple low regimens might be used to overcome tumor immunosuppression and improve the efficacy and outcome of antitumor immune responses in a mouse model.
Nazanin Nazari; Shirin Farjadian
Volume 13, Issue 3 , September 2016, , Pages 178-185
Abstract
Background: HLA-G is a nonclassical HLA class I molecule which, when elevated in tumor cells, is one of the main factors involved in tumor evasion of immune responses including NK and T cells. Objective: To evaluate the effect of HLA-G downregulation on NK cell cytotoxicity in tumor cell lines. Methods: ...
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Background: HLA-G is a nonclassical HLA class I molecule which, when elevated in tumor cells, is one of the main factors involved in tumor evasion of immune responses including NK and T cells. Objective: To evaluate the effect of HLA-G downregulation on NK cell cytotoxicity in tumor cell lines. Methods: The expression level of HLA-G was measured by real-time PCR and flowcytometry after transfection of SKOV3 by shRNA.1, which targets specific sequences in exon 4, or shRNA.2, which targets both exons 4 and 6. NK-92MI cell cytotoxicity against transfected or untransfected target cell lines was measured with the lactate dehydrogenase (LDH) release assay. The Jeg-3 cell line was used as a positive control. Results: Membrane-bound HLA-G expression levels decreased significantly in both cell lines after transfection with both shRNAs compared to their corresponding untransfected cells (p<0.05). Jeg-3 cells were better lysed than SKOV3 cells by NK cells during the first 48 h after transfection with both shRNAs. Compared to untransfected cells, shRNA.1-transfected SKOV3 cells were significantly more lysed by NK cells 24 h post-transfection (p=0.043). Conclusion: As a clinical approach, HLA-G downregulation with shRNA may be effective in cancer therapy by improving immune cell activation.
Neda Mousavi Niri; Mansooreh Jaberipour; Mahboobeh Razmkhah; Abbas Ghaderi; Mojtaba Habibagahi
Volume 6, Issue 4 , December 2009, , Pages 186-194
Abstract
Background: Several studies have demonstrated the immunosuppresive effects of mes-enchymal stem cells (MSCs) in allogeneic or mitogenic interactions. Cell-cell contact inhibition and secretion of suppressive soluble factors have been suggested in this re-gard. Objective: To investigate if adipose derived ...
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Background: Several studies have demonstrated the immunosuppresive effects of mes-enchymal stem cells (MSCs) in allogeneic or mitogenic interactions. Cell-cell contact inhibition and secretion of suppressive soluble factors have been suggested in this re-gard. Objective: To investigate if adipose derived MSCs could inhibit Jurkat lym-phoblastic leukemia T cell proliferation during coculture. Methods: Adherent cells with the ability of cellular growth were isolated from normal adipose tissues. Initial charac-terization of growing cells by flow cytometry suggested their mesenchymal stem cell characteristics. Cells were maintained in culture and used during third to fifth culture passages. Jurkat or allogeneic peripheral blood mononuclear cells (PBMCs) were la-beled with carboxy fluorescein diacetate succinimidyl ester and cocultured with increas-ing doses of MSCs or MSC culture supernatant. Proliferation of PBMCs or Jurkat cells under these conditions was assessed by flow cytometry after 2 and 3 days of coculture, respectively. Results: Results showed the expression of CD105, CD166 and CD44, and the absence of CD45, CD34 and CD14 on the surface of MSC like cells. Moreover, ini-tial differentiation studies showed the potential of cell differentiation into hepatocytes. Comparison of Jurkat cell proliferation in the presence and absence of MSCs showed no significant difference, with 70% of cells displaying signs of at least one cell division. Similarly, the highest concentration of MSC culture supernatant (50% vol/vol) had no significant effect on Jurkat cell proliferation (p>0.6). The same MSC lots significantly suppressed the allogeneic PHA activated PBMCs under similar culture conditions. Conclusion: Using Jurkat cells as a model of leukemia T cells, our results indicated an uncertainty about the suppressive effect of MSCs and their inhibitory metabolites on tumor or leukemia cell proliferation. Additional systematic studies with MSCs of differ-ent sources are needed to fully characterize the immunological properties of MSCs be-fore planning clinical applications.
Reza Amani; Amir Abbasnezhad; Eskandar Hajiani; Bahman Cheraghian; Zahra Abdoli; Razieh Choghakhori
Abstract
Background: Given the variations in clinical presentation and physiopathological mechanisms in irritable bowel syndrome (IBS) subtypes, it is an acknowledged fact that the response to treatments can be disparate. Objective: To assess the effect of vitamin D on inflammatory cytokines (IL-17, IL-10, TNF-α), ...
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Background: Given the variations in clinical presentation and physiopathological mechanisms in irritable bowel syndrome (IBS) subtypes, it is an acknowledged fact that the response to treatments can be disparate. Objective: To assess the effect of vitamin D on inflammatory cytokines (IL-17, IL-10, TNF-α), and biomarkers of oxidative stress (total antioxidant capacity (TAC), and malondialdehyde (MDA)) among IBS patients. Methods: A double-blind, randomized, placebo-controlled 6-month intervention study was carried out on 90 IBS patients (85 were analyzed), as defined by the Rome III criteria. Study participants were randomly assigned to receive either 50,000 IU vitamin D3 or a placebo fortnightly. Results: Vitamin D supplementation significantly reduced the IL-17 and MDA serum levels (P<0.05) and observably increased the TAC and IL-10 serum levels (P<0.05), compared with the placebo group. Comparing different bowel habit subtypes, we observed that it was only in diarrhea predominant IBS (IBS-D) that vitamin D supplementation was able to significantly reduce the serum levels of TNF-α and IL-17 (P<0.05). However, in all subtypes, IL-10 and TAC increased, while MDA decreased (P<0.05) in vitamin D group, compared to the placebo group. Conclusion: Vitamin D3 supplementation reduces the serum IL-17 and MDA levels, and augments the serum IL-10 and TAC levels in IBS patients, particularly in IBS-D subtype. Thus, the present study demonstrates the beneficial effects of vitamin D on patients with IBS-D.
Chuling Li; Yaxiong Xu; Xianglin Luo; Fajian Chen
Abstract
Background: Group 2 innate lymphoid cells (ILC2s) promote allergic inflammation by producing interleukin-4 (IL-4), IL-5, IL-9, and IL-13. IL-18 can promote T helper 2 cell (Th2) response by inducing IL-4, and IL-13 production from mast cells and basophils. However, the regulation of IL-18 on ILC2s remained ...
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Background: Group 2 innate lymphoid cells (ILC2s) promote allergic inflammation by producing interleukin-4 (IL-4), IL-5, IL-9, and IL-13. IL-18 can promote T helper 2 cell (Th2) response by inducing IL-4, and IL-13 production from mast cells and basophils. However, the regulation of IL-18 on ILC2s remained unknown. Objective: To investigate the regulatory role of IL-18 in inducing the type 2 innate lymphoid cells. Methods: Twenty patients with allergic rhinitis (AR) and 20 controls were enrolled. The mRNA and protein levels of IL-18 in serum, as well as the frequencies of ILC2 in peripheral blood mononuclear cells (PBMCs) were measured by real-time polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and flow cytometry. The ILC2s were sorted and the mRNA expression of IL-18 receptor in ILC2 was analyzed by real-time PCR. The effects of IL-18 on the proliferation and type 2 cytokine production were detected by tritiated thymidine incorporation test, real-time PCR, and ELISA, respectively. Results: The levels of IL-18 mRNA and protein were significantly higher in AR patients than in the controls (P<0.05). The frequency of ILC2 in peripheral blood was elevated in the AR patients than in the controls. After stimulation by IL-18 and house dust mite (HDM), the expression of IL-18 receptor (IL-18R) by ILC2 was significantly up-regulated. The tritiated thymidine incorporation results showed that IL-18 promoted the proliferation of ILC2 in a dose-dependent manner. IL-18 also induced the expression of IL-5 and IL-13 proteins by ILC2. Conclusion: Our results confirmed -for the first time- the effect of IL-18 in innate immunity, which was demonstrated by direct effect on the differentiation and function of ILC2.
Reza Hosseini-Ghatar; Tahereh Soltantoyeh; Motahareh Bahadori; Jalal Khoshnoodi; Forough Golsaz-Shirazi; Mahmood Jeddi-Tehrani; Mohammad Mehdi Amiri; Fazel Shokri
Volume 14, Issue 3 , September 2017, , Pages 200-214
Abstract
Background: Human epidermal growth factor receptor 2 (HER2) has a crucial role in several malignancies. The extracellular domain of HER2 (HER2-ECD) has been extensively employed as an important target in passive and active immunotherapy. Isolated recombinant prokaryotic HER2-ECD subdomains were previously ...
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Background: Human epidermal growth factor receptor 2 (HER2) has a crucial role in several malignancies. The extracellular domain of HER2 (HER2-ECD) has been extensively employed as an important target in passive and active immunotherapy. Isolated recombinant prokaryotic HER2-ECD subdomains were previously found to be ineffective in inducing anti-tumor antibody response. Objective: To employ recombinant eukaryotic HER2-ECD subdomains to raise anti-HER2 antibodies and determine their anti-tumor activity in vitro. Methods: Two paired subdomains of HER2-ECD (DI+II and DIII+IV), representing Pertuzumab and Trastuzumab binding domains, respectively, along with the full extracellular domain of HER2 were generated in CHO-K1 cells. Polyclonal antibodies were raised against these subdomains and characterized using ELISA, flow cytometry, and immunoblot and their anti-tumor activity was assessed by XTT assay. The cross-reactivity of these antibodies was specified along with other members of the human HER family. Results: Similar to Trastuzumab and anti-HER2-ECD antibody, anti-DI+II and DIII+IV polyclonal antibodies reacted with recombinant HER2-ECD and native HER2 expressed on tumor cells. These two polyclonal antibodies were able to inhibit the binding of Pertuzumab and Trastuzumab to HER2, respectively, and did not cross-react with other members of HER family. These antibodies were able to inhibit tumor cell growth in vitro, similar to Trastuzumab. Conclusion: The high immunogenicity of human HER2 DI+II and DIII+IV subdomains in rabbits and the tumor inhibitory activity of the purified specific antibodies imply that they might be suitable for active immunotherapy in formulation with appropriate adjuvants and in combination with other HER2 specific therapeutics.
Asghar Aghamohammadi; Ali Akbar Amirzargar; Nima Parvaneh; Paul Marjousef; Mostafa Moin; Abdolhassan Farhoudi; Mehdi Yeganeh; Toshio Miyawaki
Volume 2, Issue 4 , December 2005, , Pages 201-207
Abstract
Background: The B-cell defect in X-linked agammaglobulinemia (XLA) is caused by mutations in the gene for Bruton's tyrosine kinase (BTK). BTK mutations result in deficient expression of BTK protein in peripheral blood monocytes. Methods: Using the anti-BTK monoclonal antibody (48-2H), a flow cytometric ...
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Background: The B-cell defect in X-linked agammaglobulinemia (XLA) is caused by mutations in the gene for Bruton's tyrosine kinase (BTK). BTK mutations result in deficient expression of BTK protein in peripheral blood monocytes. Methods: Using the anti-BTK monoclonal antibody (48-2H), a flow cytometric analysis of intra cytoplasmic BTK protein expression in monocytes was performed to identify Iranian patients with XLA phenotype. To examine the possible identification of XLA patients and female carriers by this assay, we studied 13 XLA families. Results: The flow cytometric assay showed deficient expression of the BTK protein in 12 (92%) families. One patient exhibited a normal level of BTK expression. The cellular mosaicism of BTK expression in monocytes from obligate carriers was clearly shown in 9 of 12 (75%) families. Conclusion: The results suggested that most XLA patients have deficient expression of the BTK protein; therefore we conclude that deficient expression of BTK protein can be evaluated by a flow cytometric assay.
Sedigheh Sharifzadeh; Helmout Modjtahedi; Mahmood Jedi Tehrani; Abbas Ghaderi
Volume 4, Issue 4 , December 2007, , Pages 206-214
Abstract
Background: Lung carcinoma is a multiple type cancer comprising of small cell and non-small cell carcinomas (NSCLC). For therapeutic and diagnostic purposes, serum monoclonal antibodies have been produced against lung cancer. Objective: To charac-terize a murine monoclonal antibody (ME3D11) reactive ...
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Background: Lung carcinoma is a multiple type cancer comprising of small cell and non-small cell carcinomas (NSCLC). For therapeutic and diagnostic purposes, serum monoclonal antibodies have been produced against lung cancer. Objective: To charac-terize a murine monoclonal antibody (ME3D11) reactive with human NSCLC. Methods: A murine monoclonal antibody (ME3D11) reactive with human NSCLC was selected after immunization of BALB/c mice with a human large cell carcinoma with neuroen-docrine differentiation, and was tested by immunofloursence staining and Western blot analysis. Results: Our study showed that the antigen recognized by ME3D11 antibody was a cell surface antigen of 170kDa. This antigen is expressed on the cell surface of all NSCLC and a few carcinoma cell lines. In contrast, this antigen is neither expressed on the cell surface of human sarcoma, nor on the hematopoietic and normal cell lines. This anti-body had no effect on spontaneous proliferation of Mehr-80 cell line in vitro. Conclusion: High degree of binding of this monoclonal antibody to NSCLC and some other carci-noma cells warrants further studies on its potential use in diagnosis and therapy of can-cer by conjugation to drugs, toxins or radionuclides.