Isolation and purification of low molecular weight proteins from liquid culture for mycobacterium tuberculosis by chromatography

Tuberculosis [TB] is a disease caused by a bacterium called Mycobacterium tuberculosis. M. tuberculosis has different molecular weight secreted antigens. Low molecular weightproteinssecreted into the culture medium by M. tuberculosisare thought to play an important role in the development new TB diagnostic tests and new vaccines against tuberculosis. In this report, we describe isolation and purification of low-molecular-weight proteinssecreted by M. tuberculosis. Initially by biphasic medium, bacteria from Lowenstein-Jensen solid medium transferred to a Dorset-Henley liquid medium and After 6 weeks of growth, the bacteria with a 0.22 micron filters of liquid medium containing secreted proteins were isolated and the secreted proteins was precipitated by ammonium sulfate. Protein concentrations were determined by using the lowry protein assay. Then low molecular weight proteins were purified by Sephadex-G75 gel chromatography and we studied purification of low molecular weight proteins by Coomassie-Blue stained SDS-PAGE. The results showed that low molecular weight secreted proteins purified from M. tuberculosis strain DT. Also, low molecular weight proteins made up approximately 65.3% of total proteins. This study demonstrated that without break down of bacteria bodies can be purified low molecular weight secreted proteins from M. tuberculosisliquid medium by Sephadex-G75 gel chromatography

Co-administration of CpG oilgonucleotides and Chenopodium album extract reverse IgG2a/IgH1 ratios and increase IFN-gamma and IL-10 prpductions in a murine model of asthma

Asthma is a disorder of increasing severity and prevalence. Recent knowledge about the pathogenesis of asthma emphasizes its inflammatory nature. CpG oligonucleotides are a class of compounds containing motifs based on the cytosine-guanine dinucleotides [CpG-ODNs]. These motifs are suppressed in mammalian DNA. They induce inflammation in mammals characterized by the induction of T helper type 1 and regulatory responses. In this paper, the effect of CpG DNA co-administration with a homemade Chenopodium album [Ch.a] extract in a murine model of asthma is reported for the first time. Balb/C mice were sensitized using Ch.a. pollen allergenic extract plus CpG-ODNs intraperitoneally and were challenged with aerosolized allergen. Results measured included IL-10 and IFN-gamma cytokines as well as IgG subclasses. For this, splenocytes from mice treated with CpG/Ag or Ag alone, were cultured in the presence of antigen. The results showed that CpG ODN administered at the time of Ch.a sensitization, effectively increased cytokines and IgG2a/IgG1 ratios compared with those in mice treated with antigen or with PBS alone[P

Study of chenopodium album allergenic extract to induce allergic asthma in murine model

The incidence of allergic and asthmatic diseases has been continuously increased in both industrial and developing countries. Extracts from various known allergens are used for the diagnostic and therapeutic purposes. To investigate the effects of an extract prepared from Chenopodium album [Ch.A.] pollen to induce allergic asthma in BALB/C mice. BALB/C mice were sensitized by i.p. injection of Ch.A. extract and alum, and an intratracheal instillation of the extract. The bronchoalveolar lavage [BAL] fluids were obtained by cannulating the trachea and lavaging the lungs and examined for eosinophilia. Splenocytes were incubated with Ch.A. extract and cell supernatants were examined for IL-4 and IL-5 by ELISA. We demonstrated that Ch.A. extract treatment in mice increased serum levels of specific IgE and production of IL-4 and IL-5 from splenocytes. An airway eosinophilia was also demonstrated in mice. These results suggest that Ch.A. allergen extract is a potential agent in inducing characteristics of allergic asthma in a mouse model useful in investigational studies