Production of recombinant Human T Lymphotropic Virus type 1 Tax protein in Rosetta (DE3) bacterial host

 HTLV1 is the first detected retrovirus causing disease in human. The physiopathology of HTLV1 related diseases was mainly linked with its Tax protein characteristics. Use of mutant Tax proteins accompanied by immune regulator drugs could help treating HTLV1 associated myelopathy patients as a modulator of potent immune response against this viral protein. Since Tax protein is not commercially available, production of recombinant Tax protein was targeted for this study. Coding sequence of Tax protein (containing R222K mutation) in the pcDNA3.1(+) was digested with BamHI and XhoI restriction enzymes, and then removed and inserted into the expression vector pET32a(+) within the same cutting sites and cloned into E.coli DH5α. Recombinant vector was confirmed with enzymatic digestion, colony PCR, and sequencing of cloned gene. E.coli Rosetta (DE3) was transformed with the recombinant plasmid and the expression was induced. The expression of protein was assayed with SDS-PAGE and western blot using monoclonal antibodies against Tax and 5His epitope. Finally, antigenic characteristic of the recombinant protein was evaluated by western blotting against patient sera. Presence of Tax protein band in the SDS-PAGE and western blot was confirmed. Western blotting of the recombinant protein with patient sera showed the band related to Tax protein. The recombinant protein is well produced and could be detected by patients' sera, making it eligible to be used as a recombinant viral antigen for future purposes

A comparison of HTLV-1 Infection Frequency in Patients with or without Tuberculosis

Introduction: To recognize the predisposing factors in tuberculosis as an endemic infection in Northeast province of Iran, this study was aimed to evaluate whether HumanT-lymphocyte type 1 (HTLV-I) as an immunosuppressive factor increases the risk of tuberculosis. Materials and Methods: A Case-control study was conducted in 278 tuberculosis patients from 2007 to 2010, in Mashhad, Iran. Tuberculosis has been diagnosed by gold standard tests like sputum culture, bronchoalveolar lavage (BAL) culture or cytology. For detection of HTLV-I antibody, Enzyme Linked Immunosorbant Assay method and western Blot as the confirming test were performed. Then 276 healthy cases were matched for gender and age. Results: The mean age of tuberculosis patients was 49.67±21.36 years and for control cases was 48.36±20.74. In patients group, 114 (41.6%) were male, 160 (58.4%) were female and in controls 123 (44.6%) were male and 153 (55.4%) were female. Pulmonary tuberculosis was presented in 84.2% of the patients. The frequency of HTLV-1 was 2.9% and 3.3% in patients and controls, respectively.HTLV-I frequency was higher in male patients and it increased by age. Conclusion: Regarding to this study, HTLV-I infection is not stand-alone sufficient for increasing the risk of tuberculosis

Evaluation of the Relationship between Cystatin C Level in Whole Saliva and Chronic Periodontitis

Introduction: Chronic periodontitis is an infectious disease resulting in inflammation in tooth supporting tissues, advanced attachment loss and bone loss. Destructive process is a result of imbalance between analyzing enzymes such as MMPs and their inhibitors. This imbalance can also occur with other enzymes such as lysosomal cysteine proteinase, Katpsyn and their inhibitor such as cystatin. Cystatin C is a protein which controls activity of extracellular cysteine proteinase in inflammatory conditions. The aim of this study was to evaluate the protective role of salivary cystatin C in periodental disease. Materials & Methods: Twenty six patients with chronic periodontitis examined by a periodontist and also with a minimum pocket depth of six mm and more in at least eight locations in the mouth were selected. To collect Total non-irritating saliva samples, the spit method was used. Salivary levels of cystatin C was evaluated by ELISA method. Data were analysed by SPSS version 11.5 software.Results: The level of cystatin C in periodontally diseased subjects was higher than that of the control group, but the difference was not statistically significant (P=0.24). In the female group with control of age variant, the level of cystatin C was significantly higher in patients with periodontitis (P=0.036), whereas in male group, the difference was not significant (P=0.086). It seems that the lower periodontal destruction in female group is as a result of higher level of cystatin C.Conclusion: The level of cystatin C in whole saliva could be used as a marker in chronic periodontitis