ORIGINAL_ARTICLE
Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis
Background: The antibody response to hepatitis B surface antigen (HBsAg) controls hepatitis B virus infection. The "a" determinant of HBsAg is the most important target for protective antibody response, diagnosis and immunoprophylaxis. Mutations in this area may induce immune escape mutants and affect the performance of HBsAg assays. Objectives: To construct clinically relevant recombinant mutant forms of HBsAg and assessment of their reactivity with anti-HBs monoclonal antibodies (MAbs). Methods: Wild type (wt) and mutant (mt) HBsAg genes were constructed by site directed mutagenesis and SEOing PCR. The amplified genes were inserted into pCMV6-neo plasmid and transfected in CHO cell line. The expression of wt- and mtHBsAg was assessed by commercial ELISA assays and stable cells were established and cloned by limiting dilution. The recombinant mutants were further characterized using a panel of anti-HBs monoclonal antibodies (MAbs) and the pattern of their reactivity was assessed by ELISA. Results: Ten HBsAg mutants having single mutation within the "a" determinant including P120E, T123N, Q129H, M133L, K141E, P142S, D144A, G145R, N146S and C147S together with a wt form were successfully constructed and expressed in CHO cells. Reactivity of anti-HBs MAbs with mtHBsAgs displayed different patterns. The effect of mutations on antibody binding differed depending on the amino acid involved and its location within the ‘‘a’’ determinant. Mutation at amino acids 123 and 145 resulted in either complete loss or significant reduction of binding to all anti-HBs MAbs. Conclusion: Our panel of mtHBsAgs is a valuable tool for assessment of the antibody response to HBV escape mutants and may have substantial implications in HBV immunological diagnostics.
https://iji.sums.ac.ir/article_16821_3a3d92a1852b46787ec79b84e62c7b44.pdf
2013-09-01
127
138
"a" Determinant
HBs Ag
Monoclonal antibody
Forough
Golsaz Shirazi
1
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
AUTHOR
Mohammad
Mehdi Amiri
2
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
AUTHOR
Hamed
Mohammadi
3
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
AUTHOR
Ali
Ahmad Bayat
4
Monoclonal Antibody Research Center
AUTHOR
Azam
Roohi
5
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
AUTHOR
Jalal
Khoshnoodi
6
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
AUTHOR
Amir Hassan
Zarnani
7
Reproductive Immunology Research Center, Avicenna Research Institute, ACECR
AUTHOR
Mahmood
Jeddi-Tehrani
mahjed@avicenna.ac.ir
8
Monoclonal Antibody Research Center
AUTHOR
Gholam Ali
Kardar
9
Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
AUTHOR
Fazel
Shokri
gakardar@tums.ac.ir
10
Department of Immunology, School of Public Health, Tehran University of Medical Sciences
LEAD_AUTHOR
ORIGINAL_ARTICLE
Arteether Exerts Antitumor Activity and Reduces CD4+CD25+FOXP3+ T-reg Cells in Vivo
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.
https://iji.sums.ac.ir/article_16823_fb33a8ed49aa1ebcbfa4f9060e72c22f.pdf
2013-09-01
139
149
Arteether
Breast cancer
immunotherapy
Maryam
Azimi Mohamadabadi
1
Department of Immunology, School of Medical Sciences, Tarbiat Modares University
AUTHOR
Zuhair
Mohammad Hassan
hasan_zm@modares.ac.ir
2
Department of Immunology, School of Medical Sciences, Tarbiat Modares University
LEAD_AUTHOR
Ahmad Zavaran
Hosseini
3
Department of Immunology, School of Medical Sciences, Tarbiat Modares University
AUTHOR
Mehrdad
Gholamzad
4
Department of Immunology, School of Medical Sciences, Tarbiat Modares University
AUTHOR
Shekoofe
Noori
5
Department of Biochemistry, School of Medical Sciences, Shahid Beheshti University
AUTHOR
Mehdi
Mahdavi
6
Department of Virology, Pasteur Institute of Iran, Tehran
AUTHOR
Hamidreza
Maroof
7
Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran
AUTHOR
ORIGINAL_ARTICLE
KIR Gene Content Does Not Contribute to Susceptibility to Graves’ Disease
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.
https://iji.sums.ac.ir/article_16824_c88b62c069d4b5f94aa2cf3e3c98d5a9.pdf
2013-09-01
150
157
Graves’ Disease
KIR Gene
PCR-SSP
Elham
Ashouri
ashourie@sums.ac.ir
1
Endocrinology and Metabolism Research Center
LEAD_AUTHOR
Mohammad Hossein
Dabbaghmanesh
2
Endocrinology and Metabolism Research Center
AUTHOR
Amirhossein
Hadaegh
3
Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
AUTHOR
Soodeh
Rowhanirad
4
Endocrinology and Metabolism Research Center
AUTHOR
Marizeh
Bakhshayeshkaram
5
Endocrinology and Metabolism Research Center
AUTHOR
Gholamhossein
Ranjbar Omrani
6
Endocrinology and Metabolism Research Center
AUTHOR
ORIGINAL_ARTICLE
Interleukin-8 but Not Interleukin-6 Variant May Affect Susceptibility to Brucellosis
Background: Increased levels of interleukin-8 (IL-8) and interleukin-6 (IL-6) in acute human brucellosis have been reported. Previous studies have shown that the production and level of IL-6 and IL-8 cytokines are associated with the polymorphism of the encoding genes. Objective: To investigate the probable association between IL-6 (-174 C/G) and IL-8 (-251 A/T) gene polymorphisms and susceptibility/resistance to brucellosis. Methods: The patient group included 196 patients suffering from Brucella infection and the control group consisted of 82 healthy animal husbandmen from the same geographical area. IL-8 (-251 A/C) and IL-6 (-174 C/G) gene polymorphisms were analyzed by PCR-RFLP and Allele Specific PCR (AS-PCR) respectively. Results: The frequency of -251 IL-8 AA genotype was significantly lower in the controls compared with that of the patients (p=0.0051), while the frequencies of other genotypes (AT and TT) and alleles (A and T) were not significantly different among the participants. No association was found between IL-6 (-174 C/G) polymorphism and brucellosis. Conclusion: This study indicates that the IL-8 -251 AA genotype may be considered as a genetic susceptibility factor for brucellosis.
https://iji.sums.ac.ir/article_16830_dbadf11e717fd7887964952940ddd70d.pdf
2013-09-01
158
166
Brucellosis
Interleukin-6
Interleukin-8
Polymorphism
Sadaf
Asaei
1
Department of Immunology, Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz
AUTHOR
Manoochehr
Rasouli
rasoulim@sums.ac.ir
2
Department of Immunology, Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz
LEAD_AUTHOR
Ali
Moravej
3
Department of Immunology, Fasa University of Medical Sciences, Fasa, Iran
AUTHOR
ORIGINAL_ARTICLE
Serum IL-18 and hsCRP Correlate with Insulin Resistance without Effect of Calcitriol Treatment on Type 2 Diabetes
Background: Chronic low-grade systemic inflammation presented in Type 2 diabetes mellitus plays a major role in disease progression as well as development of micro- and macro-vascular complications of diabetes. Therefore, reducing inflammation can be beneficial in prevention of diabetes complications. Objectives: To investigate the association between insulin resistance and inflammatory markers, and assessing the effects of oral Calcitriol on inflammatory cytokines in type 2 diabetic patients. Methods: In this double-blind randomized placebo-controlled trial, 70 participants with type-2 diabetes were randomly divided to two groups. One group received two capsules of Calcitriol (0.25 μg 1,25-dihydroxy cholecalciferol per each capsule) per day. The second group received placebo tablets. At the beginning of the study, we assessed insulin resistance and its relation to inflammatory profile. Serum high sensitive Creactive protein (hs CRP), interleukin-6 and interleukin-18 were also measured at the beginning and the end of the 12-week supplementation trial. Results: Mean calcium, phosphorus and vitamin D concentrations in the study participants were 8.98 ± 0.79 mg/dl, 3.86 ± 0.50 mg/dl and 40.91 ± 30.9 ng/ml, respectively. IL-18 and hsCRP had significant positive associations with insulin resistance markers and negative associations with insulin sensitivity markers. At the end of the 12-week supplementation trial, no significant difference was seen in serum levels of hsCRP, IL-6 and IL-18 between the two groups, while these values were adjusted for baseline values. Conclusion: Inflammation was associated with insulin resistance in diabetic patients. No anti-inflammatory effect of Calcitriol in terms of decreasing hsCRP, IL-6 and IL-18 detected.
https://iji.sums.ac.ir/article_16832_9dfe3330b3e18d452d5445c7ae5d9166.pdf
2013-09-01
167
176
Type 2 Diabetes
IL-18
IL-6
Marzieh
Akbarzadeh
1
Department of Nutrition, School of Health and Nutrition
AUTHOR
Mohammad Hassan
Eftekhari
2
Department of Nutrition, School of Health and Nutrition
AUTHOR
Mohammad Hossein
Dabbaghmanesh
dabbaghm@sums.ac.ir
3
Endocrine and Metabolism Research Center, Nemazee Hospital
LEAD_AUTHOR
Jafar
Hasanzadeh
4
Epidemiology Department, School of Health and Nutrition, Shiraz University of Medical Sciences, Shiraz, Iran
AUTHOR
Marzieh
Bakhshayeshkaram
nahendo2@sums.ac.ir
5
Endocrine and Metabolism Research Center, Nemazee Hospital
LEAD_AUTHOR
ORIGINAL_ARTICLE
Interleukin-17 Gene Expression and Serum Levels in Children with Severe Asthma
Background: IL-17 is a major cytokine player in T cell mediated leukocyte associated inflammation. IL-17 is also recognized to participate in the pathophysiology of asthma. Objective: To determine the role of IL-17 in predicting severe asthma. Methods: We obtained serum samples from asthmatic children under the age of 5-year in three different groups of mild (n=33), moderate (n=28) and severe (n=32) persistent asthma. IL-17 serum concentrations and mRNA expression were determined by ELISA and real time PCR assays, respectively. Results: Serum IL-17 concentrations were significantly higher in patients with severe asthma than the other two groups of children with mild and moderate disease (p=0.00). Mean serum IL-17 values were 142.04 pg/ml in mild group, 180.4 pg/ml in moderate group and 251.25 pg/ml in severe group. IL-17 mRNA levels were also significantly elevated in severe asthmatic patients compared to mild and moderate asthmatic children (p=0.00). Conclusion: Our data reveal an increase in the serum IL-17 concentrations and IL-17 mRNA expressions in children with severe asthma compared to those with mild and moderate forms of the disease.
https://iji.sums.ac.ir/article_16834_dff4322a2b35feb8b1cd4a2969316d38.pdf
2013-09-01
177
185
Soheila
Alyasin
1
Allergy Research Center
AUTHOR
Mohammad Hossein
Karimi
karimimh@sums.ac.ir
2
Transplant Research Center
AUTHOR
Reza
Amin
3
Allergy Research Center
AUTHOR
Maryam
Babaei
4
Allergy Research Center
AUTHOR
Sepideh
Darougar
sepidehdarougar@yahoo.com
5
Allergy Research Center
LEAD_AUTHOR
ORIGINAL_ARTICLE
A Peculiar Case of Splenic Marginal Zone Lymphoma and Review of Literature
https://iji.sums.ac.ir/article_16835_bcbad4de6f7ba4caa1a0e36e9c9ce656.pdf
2013-09-01
186
189
Mahendra
Narain Mishra
mnmishra@hotmail.com
1
Department of Pathology, Dr Lal Path Labs Pvt. Ltd. National Reference Laboratory, New Delhi
LEAD_AUTHOR
Rakesh
Pandey
2
Sanjay Gandhi Postgraduate Institute of Medical Sciences, Department of Pathology, All India Institute of Medical Sciences, New Delhi
AUTHOR
Ashok
Dinda
3
Sanjay Gandhi Postgraduate Institute of Medical Sciences, Department of Pathology, All India Institute of Medical Sciences, New Delhi
AUTHOR
Soniya
Nityanand
4
Sanjay Gandhi Postgraduate Institute of Medical Sciences, All India Institute of Medical Sciences, India
AUTHOR
ORIGINAL_ARTICLE
Does Nitric Oxide Generated by Dendritic Cells Contribute to the Low Incidence of GVHD after Cord Blood Transplantation?
https://iji.sums.ac.ir/article_16851_d3c1fe195e4131693494e4e1d24a49b8.pdf
2013-09-01
190
192
Nadereh
Naderi
nnaderi@hums.ir
1
Center for Research of Molecular Medicine
LEAD_AUTHOR
Seyed Mohammad
Moazzeni
2
Department of Immunology, Tarbiat Modares University
AUTHOR
Ali Akbar
Pourfathollah
3
Department of Immunology, Tarbiat Modares University
AUTHOR
Kamran
Alimoghaddam
4
Hematology-Oncology and BMT Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
AUTHOR