Immunogenicity of HLA-DR1 restricted peptides derived from leishmania major gp63 using FVB/N-DR1 transgenic mouse model

Leishmaniasis is a worldwide disease prevalent in tropical and sub- tropical countries. In the present study the immunogenicity of three human HLA-DR1 restricted peptides derived from L. major gp63 protein was evaluated using FVB/N-DR1 transgenic mouse model. The immunity generated by three MHC class II - restricted peptides with the sequence of AARLVRLAAAGAAVT [AAR], AAPLVRLAAAGAAVT [AAP] and SRYDQLVTRVVTHE [ASR] derived from L. major gp63 protein were predicted using a web-based software [SYFPEITHI] and tested in FVB/N-DR1 transgenic mice. Immunization of FVB/N-DR1 transgenic mice with one of the three predicted peptides [AAR] resulted in high levels of Th1-type immune response as well as significant levels of IFN-gamma detected by Proliferation assay and ELISA. The results indicate a high level of immunogenicity for AAR, which can be a potent candidate for peptide vaccine in Leishmania infections.

Leishmania mexicana Gp63 cDNA using gene gun induced higher immunity to L. mexicana infection compared to soluble leishmania antigen in BALB/C

Leishmaniasis is a worldwide disease prevalent in tropical and sub tropical countries. Many attempts have been made and different strategies have been approached to develop a potent vaccine against Leishmania. DNA immunisation is a method, which is shown to be effective in Leishmania vaccination. Leishmania Soluble Antigen [SLA] has also recently been used Leishmania vaccination. The immunity generated by SLA and L. mexicana gp63 cDNA was compared in groups of 6 mice, which were statistically analysed by student t- test with the P-value of 0.05. SLA was administered by two different methods; intramuscular injection and injection of dendritic cells [DCs] loaded with SLA. L. mexicana gp63 cDNA was administered by the gene gun. Immunisation of BALB/c mice with L. mexicana gp63 resulted in high levels of Th1- type immune response and cytotoxic T lymphocytes [CTL] activity, which were accompanied with protection induced by the immunisation against L. mexicana infection. In contrast, administration of SLA, produced a mixed Th1/Th2-type immune responses as well as a high level of CTL activity but did not protect mice from the infection. The results indicate higher protection by DNA immunisation using L. mexicana gp63 cDNA compared to SLA, which is accompanied by a high level of Th1 immune response. However, the CTL activity does not necessarily correlate with the protection induced by the vaccine. Also, gene gun immunisation is a potential approach in Leishmania vaccination. These findings would be helpful in opening new windows in Leishmania vaccine research

population-based seroepidemiological study on hepatitis E virus in Iran

Little is known about HEV seroprevalence and its determinants in Iran. Considering the fact that Iran is among the countries in which HEV infection is endemic, a large-scale population-based study in this regard is justified. This survey was conducted in 2006 in Tehran and Golestan Provinces, Iran. Stored sera of subjects were tested for serological markers of anti-HEV. The baseline data were recorded in structured questionnaires. Weighted seroprevalence and weighted logistic regression coefficients were calculated. A total of 1423 samples were included. The overall seroprevalence in two provinces was 7.4%. Age with an odds ratio equal to 1.59 [95% CI: 1.26-2.02] and history of traditional phlebotomy with an odds ratio equal to 2.28 [95% CI: 1.13-4.60] were independent predictors of HEV seropositivity. Considering the high rate of HEV seroprevalence in Iran, further studies on the cost-effectiveness of vaccination among vulnerable groups are mandator

Prevalence of non alcoholic steatohepatitis in Iran: a population based study

Non-alcoholic fatty liver disease/steatohepatitis [NAFLD/NASH] is the most common form of chronic liver disease woldwide and is no longer considered a benign disease. Its prevalence has not been determined in a large-scale population-based study in Iran. A total of 6583 individuals aged 18 to 65 were randomly selected from three geographically distinct provinces in Iran. Blood samples were obtained from each subject and a questionnaire was completed exploring data including self-admitted regular alcohol use. Serums were tested for anti-HCV antibody [anti-HCV], hepatitis B surface antigen and anti-hepatitis B core antibody. Positive samples for anti-HCV antibody were re-tested and those positive in a repeat ELISA were confirmed by a recombinant immunoblot assay [RIBA] test. Serums were also tested for ALT levels. Subjects with elevated ALT defined as serum ALT >/= 40 IU/L with no history of alcohol consumption and negative HBV and HCV infection were considered as [presumed NASH]. In this study 5589 subjects were analyzed. Two hundred and forty two individuals [4.3%] were diagnosed with elevated ALT levels. Among individuals with elevated ALT, 15 [6.2%] were diagnosed with either hepatitis B or hepatitis C. The overall weighted prevalence of presumed NASH was 2.9%. According to multivariate analysis, male sex, urban lifestyle, and being overweight or obese were significantly associated with [presumed NASH]. Obesity and metabolic syndrome, the most predictive factors of fatty liver disease, are increasing in Iran, therefore the prevalence of NAFLD/NASH and related complications are expected to increase in the future. This population based study gives a crude estimate of the prevalence of NASH around the country. Studies with more accurate surrogates of NASH need to be done. The disparity among different provinces merits special consideration

Isolation, cloning, expression and purification of recombinant RhD antigen from cord blood

Rh [Rhesus] is a highly complex blood group system in man deeply rooted in transfusion medicine. Isolation of RhD from cord blood, cloning and expression of recombinant RhD antigen in bacterial expression system was the aim of this study. Total RNAs were extracted from cord blood [O[+]]. The quality of RNA was determined by electrophoresis. In order to obtain coding sequence of RhD antigen cDNA was synthesized and Rh D gene was amplified by RT-PCR. The isolated RhD gene was cloned to pUCIS vector and transformed to DH5alpha. The confirmed construct was sub cloned into expression vector, pBADgIII/A, and expressed in Top 10 E.coli. The expressed protein was characterized by SDS-PAGE and western blot analysis. Antigenicity of the expressed protein was assessed by ELISA using commercially available human anti-RhD polyclonal antibody with peroxidase conjugated goat anti-human IgG, IgM, IgA as secondary antibody. RhD gene was successfully cloned and expressed. The expected size of recombinant RhD protein was detected in SDS-PAGE, and confirmed by dot and western blot analysis. RhD antibody reacted with recombinant RhD antigen as well as with RhD polypeptide extracted from RBCs membrane. The recombinant RhD may be helpful to further investigate the molecular basis of RhD protein and could be applicable for production anti- D antibody in an animal model

preliminary report on prevalence of antibody response to GBV-C, E2 protien in Iranian blood donors and multitransfused patients

To analyze serologic markers of Hepatitis G virus [GBV-c] infection in Iranian blood donors and two major groups of multitransfused patients, hemophiliacs and thalassemics. Nine hundred and five serum samples collected from the volunteer blood donors and two patient groups under the study were tested for the presence of antibodies to the GBV-C antigen [anti E2] by an ELISA assay .Those found positive for anti- E2 were also tested for possible exposure to HCV by detecting anti- HCV in their sera. Levels of ALT were also tested to evaluate impact on liver function. Approximately 8.6% of the volunteer blood donors were found positive for anti-E2 .The prevalence rate in hemophiliacs was 41.4% and in thalassemia patients was 33.4%, which was significantly [P< 0.05] higher than donors. However, the prevalence rate among the two high risk groups was not statistically significant. A large number of the healthy blood donors in Iran have been exposed to the GBV-C. The significantly higher levels seen in the multitransfused patients can be regarded as an important route of transmission. It seems that no evidence of liver damage in individuals exposed confirming that GBV-C is not a hepatitis virus

[Hepatitis A and E in the east of Golestan province]

Hepatitis A and E are among the most important etiological factors of acute hepatitis, which can lead to fulminant liver disease and death. The aim of this study was to determine the prevalence of hepatitis A and E in the east of Golestan Province, Iran. Eight hundred inhabitants of Gonbad City and Ghamar Ghozi Village from Kalale district were randomly selected for the study. Hepatitis A virus Antibody [Anti HAV] and Hepatitis E virus Antibody [Anti HEV] were checked by ELISA method. A total of 697 subjects [441 females] with a mean age of 43 +/- 15.1 years participated in this study. A total of 687 [98.6%] and 82 [11.8%] persons had positive Anti HAV and Anti HEV, respectively. This research is the first study on the prevalence of hepatitis A and E in the general population of Iran, which shows that HA V infection is pandemic and hepatitis E is common in the area

[Hepatitis C in Golestan province-Iran]

Hepatitis C is one of the most important etiological factor in chronic liver disease which could lead to the progressive liver cirrhosis and hepatocellular carcinoma. The purpose of this study is to determine the prevalence of Hepatitis C in east of Golestan province [north-east of Iran]. 2123 inhabitants from north-east of Iran of general population have been studied using randomized selection. The taken samples were fresh serum searching for Hepatitis C Virus Antibody [HCV Ab] via ELISA method. A recombinant immunoblot assay [RIBA] was done for the positive HCV Ab samples tested with ELISA as the next step. A total of 2123 samples [female: 1387] with mean age of 41.3+/- 14.8 years and 960 rural and 1141 urban were studied. Fifty six samples [female:44] were HCVAb positive [ELISA], twenty of fifty six samples were also RIBA positive [female: 18 equal 81.8% and male: 4 equal 18.2%]. According to ELISA result the prevalence of HCVAb positive samples in female was 3.1% [44 of 1387 samples] and in male was 1.6 [12 of 714 samples]. As a total result we reached out that the prevalence of Hepatitis C through ELISA method was around 2.6%. This number reduced to 1% considering RIBA positive samples which is the definite test in Hepatitis C diagnosis. This study is the first Hepatitis C determination in general population in Iran. A comparison between the Hepatitis C prevalence in north east Iran and other part of the world showed that Iran relatively has a lower prevalence [Asia: 3.55% Africa: 5.17% America: 1.9%]. No other Hepatitis C evaluation study has been done in any other part of Iran

Expression of recombinant human serum albumin in the methylotrophic yeast; Hansenula polymorpha

Human serum albumin [HAS] is the major protein component of human plasma. It plays a very important role in transporting of macro molecules and maintaining the normal osmolarity. It is used as a therapeutical protein in patients with hypoalbuminemia and acute bleeding and burning. Albumin consumption in the world is about 500 ton/year. The aim of this research is to study the production of rHSA in shake flask culture by Hansenula polymorpha. H. polymorpha was used for the production of recombinant human serum albumin [rHSA] in several of shake flask culturing; expression of rHSA was investigated relating several parameters affecting the expression of HSA. To optimize the secretory expression of rHSA under the control of FMD promoter in H polymorpha RB-11 incubation time, culture media temperature and protease inhibitors were analyzed. This study not only established production of rHSA in yeast but also analyzed the correlation between affecting parameters and the level of HSA expression. Comparison of the HSA levels in the culture supernatants showed that the highest HSA yield was 17.6mg/l. The research shows that among three different temperatures [25°C,30°C and 37°C] 37°C was the best temperature and amongst three different incubation times [24h,48h and 72h] 48h was the optimum time and YNB 1% glycerol with buffer was the best derepression medium in comparison with others. Using these optimized conditions, stable production of rHSA of around 17.6mg/l was achieved. Our results suggest that affecting experssion factors improved in this study are suitable for production of recombinant albumin

Improving purification of coagulation F IX using heparin affinity chromatography and its comparison with ion exchange chromatography

Hemophilia B is a genetic disorder due to deficiency or complete absence of factor IX coagulation factor. Treatment of choice for these patients is use of factor IX concentrates. Therefore, purification of plasma proteins and separation of factor IX have been major objectives for scientists involved in this field. In this respect, purification procedure using ion exchange chromatography is widely used, but in the past decade affinity chromatography was also introduced. The objective of the present study has been to apply both techniques for the purification of factor IX and compare the quality and yield of the product. For the purification procedure, chromatography columns [XK-16], containing DEAE sepharose and Heparin sepharose were used. Factor IX coagulation activity was measured using a one-stage coagulation assay and factor IX antigen was quantified using ELISA technique. The specific activity and relative increase in purity of factor IX was calculated and it was demonstrated that specific activity improved from 3.1 IU/mg using DEAE ion exchange to 29 IU/mg when affinity chromatography was added and purity was increased from 155 to 1450 respectively. The present study demonstrates that addition of an affinity chromatography step using heparin sepharose is a major improvement in the purification of factor IX, where both specific activity and purity are increased considerably

preliminary study on the effect of two therapeutic procedures on the expression of BCR-ABL gene in CML patients

Chronic myelogensis leukemia [CML] is a chronic myeloproliferative disorder resulting from a specific mutation in a pluripotent stem cell. The association of Philadelphia chromosome with this disorder was described in 1973. Subsequently, the BCR-ABL fusion gene and its product which is a tyrosin kinase inhibiting apoptosis were introduced. The treatment of choice for these patients is BMT as well as the new molecular treatment of imatinib mesylate. The present study was designed in order to compare the rate of expression of the BCR-ABL gene in patients who had undergone treatment by bone marrow transplantation with those treated by imatimib. Expression of BCR-ABL gene was measured in 34 patients 17 patients were treated with BMT and 17 with imatinib mesylate using quantitative PCR on a light cycler instrument. The results obtained in this preliminary study demonstrates that rate of gene expression patients treated with imatinib mesylate for more than 8 month was similar to that found in patients treated with 8 BMT who were in relatively stable conditions. This finding may be an indication that imatinib mesylate as a molecular inhibitor can have the same effect as BMT but with less adverse effects. Clearly definite conclusions require more extensive studies on larger number of patients

Iranian blood transfusion, donor safety program: Effect of long-term plasmapheresis on plasma proteins

Plasmapheresis is a well-recognised method for harvesting plasma. It was introduced in the 1950s and is currently employed worldwide. This method has been in use in Iran since 1979. To evaluate the efficacy of donor safety program used at the Iranian Blood Transfusion Organization. A comparison is made of the protein concentrations between the first-time and long-term plasmapheresis donors, using protein gel electrophoresis. The effect of vaccination for collection of hyper-immune plasma was also studied. Results/Our data comprising a 10-year evaluation of the current programmes, demonstrates no significant difference between the groups studied. This indicates safety of the plasmapheresis procedures in plasma procurement

study on the application efficacy of solvent-detergent [S/D] treatment in the process of purifying factor VII from prothrombin complex

The risks of transmitting viral infection by blood and plasma-derived products have long been known and still remain an area of concern. In this study, in the process of purifying human factor VII from prothrombin complex, S/D treatment using tri-n-butyl phosphate and Tween 80 was employed and its capability and efficacy was studied. The results indicated that the process did not affect the biological function of the essential coagulation factors studied. In addition, the process was shown to be effective on enveloped viruses, where its inactivation factor for the model used, Herpes Simplex type I, was 5.5 logs. However, the procedure was not effective on poliovirus which was used as a model for non-enveloped viruses. It is therefore concluded that for increasing viral safety it is best to apply at least one more inactivating procedure which will be effective on non-enveloped viruses as well

preliminary study on the prevalence of anti-HIV in Iran

In order to determine the prevalence of acquired immunodeficiency syndrome [AIDS] in Iran, sera obtained from different high-risk groups, as well as a number of healthy blood donors were studied for anti-HIV by enzyme immunoassay technique. It was demonstrated that the only group at risk in Iran at present are hemophiliacs who had received factor VIII concentrate imported from Western countries of whom 14.2% were found to be anti-HIV positive. Volunteer blood donors and even drug abusers did not show seropositivity

Prevalence of e-antigen and antibody among healthy blood donors carrying hepatitis B surface antigen in Iran

The prevalence of Hepatitis B "e" antigen [HBeAg] and its antibody [anti-HBe] was studied in 1228 apparently healthy blood donors carrying Hepatitis B surface antigen [HBsAg]. The frequency of HBeAg was found to be 9.4% and anti-HBe was 82%. The prevalence of HBeAg was higher in younger donors while anti-HBe was more frequent in the older ones