History and Objectives: Extraction and purification of polysaccharide capsule is the first step for preparation preventive vaccines in infections as a result of encapsulated Staphylococcus aureus. This study was carried out to extract and purify Staphylococcus aureus type 2.

Materials and Methods: The descriptive and explorative strategy of this study was conducted on Staphylococcus aureus (Smith diffuse variant) in BHI broth in a shaking incubator (200 rpm) in an aerobic condition at a temperature of 37°C. The capsule was extracted using dilute acid and heating method. Then, dialysis was done overlight at 4°C and the product was lyophilized. Lyophilized powder was purified using a cellulose DEAE column (2.6×50 cm) by gradient method using bicarbonate ammonium buffers (0.01-1 M) at a velocity of 60 mt/h (Ion exchange chromatography). Finally absorption spectrum was plotted and purified capsule was analyzed using scanning spectrophotometer.

Results: Polysaccharide capsule was collected in final growth phase at 0.1 HCL and at a temperature of 100°C. Absorption spectrum were evaluated using H2SO4/galacial acetic acid assay at wavelength range of 350-620 nm. The result showed that chromatographs are well matched with previously purified antigens.

Conclusion and Recommendations: It is concluded that it is possible to extract and purify the capsule of Staphylococcus aureus type 2 and it is recommended to produce the related preventive vaccines.

Background: Finding suitable tumor markers with higher sensitivity and specificity for malignancy could be a fine approach to early diagnosis, response to treatment and follow up after treatment in cancers. The aim of this study was to compare the sensitivity and specificity of telomerase enzyme with carcinoembryogenic antigen (CEA), Cyfra21-1 and parathyroid hormone (PTH), in diagnosis of patients with lung cancer and other cancerous diseases visited in Lung and Tuberculosis Research Center of Tabriz Medical University.

Materials and Methods: In this research a diagnostic value was designed on 50 lung cancer patients and 20 normal individuals. Telomerase activity was measured by TRAP assay, based on PCR- ELSA, in lung tumor biopsies. The serum levels of PTH, CEA, and Cyfra-21-1 were measured using commercially available immunoassay kits. Sensitivity, specificity and other diagnostic value markers were calculated. To compare quantitative means of the two groups, T independent and Man-Whitney analysis were applied.

Results: Telomerase activity had the highest sensitivity and accuracy (76% and 82.9%, respectively). The mean values for Cyfra-21-1 were 58% and 70%, respectively. The highest value of sensitivity for telomerase was calculated in small and large cell carcinoma with 100% in lung cancer patients. The highest sensitivity of Cyfra-21-1 (98%) was seen in the large cell carcinoma.

Conclusion:  The diagnostic sensitivity of telomerase enzyme was higher than that of cyfra21-1 and CEA. It seems that the positive value of telomerase enzyme could be considered as a rapid diagnostic factor especially in lower stages of lung cancer.

Background: Considering the immunosuppressive effects and prevalent mutations in some HCV antigens, induction of CD8+ T cell responses is focused on conserved and critical epitopes which as a multi-epitope vaccine can prevent the chronic nature of the disease.

Materials and Methods: Two immunodominant HLA-A2-restricted human epitopes (E2614-622 and NS31406-1415) and two H-2d-restricted mouse epitopes (core132-142 and E2405-414) were designed in a sequential tandem, predicted by immunoinformatic analyses. Following the synthesis, related nucleotide sequence was cloned into the pcDNA3.1 vector with and without the fusion of hepatitis B surface antigen (HBsAg). Two constructed plasmids (pcDNA3.1.HPOL and pcDNA3.1.POL, respectively) were evaluated for the protein expression and secretion in Cos-7 cell line. After the vaccination of BALB/c mice (n=6 in each group) with different DNA and peptide immunization regimens, CD8+ T cell activity as well as the type and protective potency of the induced responses were evaluated.

Results: Despite the induction of epitope-specific responses in pcDNA3.1.POL injected mice, the group immunized with pcDNA3.1.HPOL indicated a significant increase in the number and activity of CD8+ T cells (P<0.05). Peptide boosting of this group (formulated in two human-compatible adjuvant) still led to the more activation of CD8+ cells, induction of Th1 response and the inhibition of tumor model growth (P<0.05).

Conclusion: Fusion of HBsAg as a particle-forming sequence and the source of helper epitopes along the DNA-prime/peptide-boosting immunization regimen are proposed as two promising strategies to improve the CTL multi-epitope vaccines against HCV.

Background: Inherited thrombophilic gene polymorphisms have been related to the pathogenesis of venous thromboembolism and its outcomes. Considering the scarcity of data on the frequency of the thrombophilic gene polymorphisms in Iranian populations, the aim of this study was to assess such polymorphisms in healthy individuals.

Materials and Methods: This cross-sectional study was performed on 304 healthy blood donors with no history of venous thromboembolism in Shahrekord. Venous blood was collected in EDTA-treated tubes and then, genotyping of the factor V Leiden, prothrombin G20210A, MTHFR C677T and PLA2 polymorphisms was done using PCR – RFLP.

Results: Six (1.97 %) cases were heterozygous for factor V Leiden and one was homozygous. Ninty-four (30.92%) and 11 (3.62%) subjects were heterozygous and homozygous for MTHFR C677T, respectively. Two (0.6%) cases were heterozygous for prothrombin G20210A and there was no homozygous case. Twenty-eight (9.2%) and 2 (0.6%) cases were heterozygous and homozygous for PLA2, respectively. In addition, 44.6% of the study population and 14.5%, with the deletion of MTHFR C677T, carried at least one thrombophilia polymorphism.

Conclusion: The frequency of thrombophilia polymorphisms is different from the previously published data in Caucasians and also the limited existing data in Kermanshah (Iran). Moreover, the discrepancies may be associated with the ethnic differences and sample selection.

Background: Severe or lethal damages of toxoplasmosis clearly indicate the need for the development of a more effective vaccine. Immunization with recombinant plasmid encoding protective proteins is a promising vaccination technique. Therefore, this study aimed to evaluate the immunization with plasmid encoding GRA5 antigen of Toxoplasma gondii in BALB/c mice.

Materials and Methods: In this experimental study, three groups of BALB/c mice (n=10 in each group) were selected using simple random sampling. GRA5 gene was cloned into pcDNA3 plasmid and purified by plasmid purification kits and then the product was injected (IM). To determine the status of cellular and humoral immunity, the Il-4, IFN-y and IgG, IgG2a, IgG subtypes were evaluated respectively using the ELISA-based assay.

Results: The group immunized with pcGRA5 indicated a significant augmented response in humoral and cellular immunity (P≤0.05) which was confirmed by MTT test. The mean survival time for the experimental and control groups were 9 and 6 days, respectively.

Conclusion: The immunized mice by pcGRA5 produce the higher titers of IFNγ indicated a Th1 response which is confirmed by the high level of IgG2a. Findings of this study demonstrate that GRA5 gene of T. gondii can be a potential vaccine candidate against the toxoplasmosis.

Background: Cytotoxic lymphocyte antigen-4 (CTLA-4) plays an important role in regulating T cell activation. CTLA-4 gene polymorphism is related to genetic susceptibility to various autoimmune diseases, including systemic lupus erythematosus (SLE). This study aimed to evaluate the role of CTLA-4 polymorphism at positions-1661AG in patients with SLE.

Materials and Methods: This study was performed on 180 SLE patients referred to 5th Azar educational hospital (Gorgan, Iran, during 2010-2011) and 304 ethnically-and age-matched healthy controls. Polymerase chain reaction restriction fragments length polymorphism (PCR-RFLP) was used to analyze the genotype and allele frequencies of this polymorphism.

Results: No statistically significant difference was observed between the studied genotypic and allelic frequencies between the SLE patients and healthy controls. Moreover, no significant correlation was found between the different risk factors (e.g., age, ethnicity, history of the disease and the parents' relationship) and the different genotypes.

Conclusion: Results suggest that the -1661AG polymorphism in the promoter region of the CTLA-4 gene has no role in the genetic susceptibility to SLE.