Original Article
Volume 5, Issue 3 , September 2008, Pages 148-148
Original Article
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.
Original Article
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.
Original Article
Arash Mahboubi; Mohammad Reza Fazeli; Rasoul Dinavand; Nasrin Samadi; Mohammad Sharifzadeh; Houshmand Ilka; Saeed Azadi; Hassan Kalkouei; Rasoul Hajikhanmirzaei; Mahboubeh Valadkhani
Volume 5, Issue 3 , September 2008, Pages 163-170
Abstract
Background: Several adjuvants have been evaluated for vaccine formulations but alu-minum salts will continue to be used for many years due to their safety, low cost and adjuvanticity with different antigens. Two commonly used aluminum adjuvants, alumi-num hydroxide and aluminum phosphate have different ...
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Background: Several adjuvants have been evaluated for vaccine formulations but alu-minum salts will continue to be used for many years due to their safety, low cost and adjuvanticity with different antigens. Two commonly used aluminum adjuvants, alumi-num hydroxide and aluminum phosphate have different adjuvanticity properties. Com-mercial recombinant protein hepatitis B vaccines containing aluminum hydroxide is fac-ing low induction of immunity in some sections of the vaccinated population. Objec-tive: In this study, to follow the current global efforts in finding more potent hepatitis B vaccine formulations, adjuvanticity of aluminum phosphate, aluminum hydroxide and their combinations has been evaluated. Methods: The formulated vaccines were admin-istered intra-peritoneally (i.p.) to BALB/c mice and the titer of antibody was determined after 28 days using ELISA technique. The geometric mean of antibody titer (GMT, mIU/ml), seroconversion and seroprotection rates, ED50 (ng) and relative potency (μg/dose) of different formulations were determined. Results: GMT of antibody titer, seroconversion and seroprotection rates showed significantly higher adjuvanticity for aluminum phosphate than other formulations. The ED50 of aluminum phosphate was approximately two fold less than other formulations. Conclusion: Aluminum phosphate showed more adjuvanticity than aluminum hydroxide and their combinations in hepati-tis B protein vaccine. The use of aluminum phosphate as adjuvant leads to higher im-munity which may result in more protective response in vaccinated groups.
Original Article
Shirin Farjadian; Mehrdad Lotfazar; Abbas Ghaderi
Volume 5, Issue 3 , September 2008, Pages 171-176
Abstract
Background: Papillon-Lefevre syndrome (PLS) is a rare autosomal recessive disorder characterized by palmoplantar hyperkeratosis and early development of aggressive pe-riodontitis. Although cathepsin C (CTSC) gene mutations have been established in about 70-80% of PLS patients, it is assumed that the ...
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Background: Papillon-Lefevre syndrome (PLS) is a rare autosomal recessive disorder characterized by palmoplantar hyperkeratosis and early development of aggressive pe-riodontitis. Although cathepsin C (CTSC) gene mutations have been established in about 70-80% of PLS patients, it is assumed that the patients may have dysfunctioning of immune defense mechanisms. Objective: To assess the association of HLA class II genes and PLS. Methods: HLA class II genes were typed in nine Iranian PLS patients and their family members and the results were compared to 816 Iranian healthy sub-jects. Results: The results of this study revealed that DRB1*0101 and DRB1*0301 al-leles were more frequent in PLS patients than in normal controls. However, there was no significant difference between PLS patients and normal controls. Moreover, the same haplotypes and genotype combinations were also observed in some patients and their healthy siblings. Conclusion: The results of this study showed no strong association between HLA class II alleles and PLS.
Short Paper
Kazem Ahmadi; Majid Riazipour
Volume 5, Issue 3 , September 2008, Pages 177-180
Abstract
Background: T-2 toxin is a mycotoxin of type A trichothecenes produced by several fungal genera such as Fusarium species. Mycotoxins can affect both cell mediated and humoral immune compartments. Objective: The purpose of this study was to investi-gate the effect of T-2 toxin on cytokine production by ...
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Background: T-2 toxin is a mycotoxin of type A trichothecenes produced by several fungal genera such as Fusarium species. Mycotoxins can affect both cell mediated and humoral immune compartments. Objective: The purpose of this study was to investi-gate the effect of T-2 toxin on cytokine production by mouse peritoneal macrophages and lymph node T cells. Methods: Mouse peritoneal macrophages and lymph node T cells were isolated and treated with different concentrations of T-2 toxin and incubated at 370C and 5% CO2 in air for 48 hours. Cell free media were removed and used for cy-tokine assay by an ELISA method. Results: T-2 toxin significantly reduced IL-1β re-lease in a concentration dependent manner (p<0.005, p<0.001). Interleukin-12 and TNF-α production were significantly increased in response to 0.001ng/ml, 0.01ng/ml and 0.1ng/ml of T-2 toxin (p<0.001). However, T-2 toxin at higher concentrations rang-ing from 1ng/ml to 100ng/ml, reduced both IL-12 (p<0.001) and TNF-α production (p<0.005, p<0.05). The effects of T-2 toxin on lymph node T cells showed that IL-4 and IL-10 release was decreased in a concentration dependent manner (all with p<0.01). T-2 toxin at concentrations between 1ng/ml and 100ng/ml reduced the release of both IL-2 and IFN-γ (p<0.05, p<0.001). Conclusion: The results suggest that T-2 toxin at low concentrations can highly induce secretion of IL-12, TNF-α, IFN-γ and IL-2 and it may be used as a positive immunomodulator in the human model.
Short Paper
Sara Kashef; Farid Ghazizadeh; Ali Derakhshan; Shirin Farjadian; Soheila Alyasin
Volume 5, Issue 3 , September 2008, Pages 181-184
Abstract
Background: Infection is now the most common cause of morbidity in Systemic Lupus Erythematosus (SLE). There is lack of information regarding the specific antibody forma-tion in response to vaccines in young SLE patients. Objective: To determine the efficacy of anti-tetanus antibody response in young ...
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Background: Infection is now the most common cause of morbidity in Systemic Lupus Erythematosus (SLE). There is lack of information regarding the specific antibody forma-tion in response to vaccines in young SLE patients. Objective: To determine the efficacy of anti-tetanus antibody response in young patients with SLE. Methods: Forty SLE pa-tients with mean age of 14.1 years (range: 7-21) and 60 age and sex matched normal con-trols were enrolled in this study over a period of one year. Diagnosis was made according to the ACR criteria and disease activity was determined based on SLE Disease Activity Index (SLEDAI). All patients and controls had received the complete schedule of tetanus vaccinations consisting of three primary doses and two boosters by the age of six. Serum immunoglobulins and anti-tetanus antibody titers were determined by Nephelometry and ELISA. Anti-tetanus antibody levels greater than 0.1 IU/ml have been suggested as pro-tective. Results: In all of the patients and controls anti-tetanus antibody titer was > 0.1 IU/ml. IgG, IgA, and IgM levels were in the normal range for their age. Mean disease ac-tivity score was 4.9 (range: 0-16). There was no association between SLEDAI score and anti-tetanus antibody response. Conclusion: School age onset and immunosuppressive therapy does not seem to interfere with development of consistent immunity to tetanus vaccine in young SLE patients.
Short Paper
Khalid Huseein Bakheit; Neda Kamal Bayoumi; Ishagh Adam
Volume 5, Issue 3 , September 2008, Pages 185-188
Abstract
Background: Cesarean section delivery can lead to much maternal morbidity. Different cytokines have been reported to be influenced by the mode of delivery. Objective: To investigate the influence of mode of delivery on maternal, placental and cord sera of interferon gamma (IFN-γ), interleukin-4 ...
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Background: Cesarean section delivery can lead to much maternal morbidity. Different cytokines have been reported to be influenced by the mode of delivery. Objective: To investigate the influence of mode of delivery on maternal, placental and cord sera of interferon gamma (IFN-γ), interleukin-4 (IL-4) and interleukin-10 (IL-10) levels. Methods: These three cytokines were measured using ELISA in peripheral, placental and cord sera of two groups of women (38 in each group), either delivering vaginally or by elective cesarean section. Results: Concentrations of IFN-γ, IL-4 and IL-10 in the peripheral and placental sera were higher in vaginal delivery, while cord cytokines were not significantly different in the two groups. Cord sera contained significantly less con-centrations of these cytokines than the peripheral and placental sera. Conclusion: It ap-pears that the levels of IFN-γ, IL-4 and IL-10 are influenced by the mode of delivery.
