Immunotherapy of chenopodium album induced asthma by intranasal administration of CpG oligodeoxynucleotides in BALB/c mice

There are many therapeutic methods for allergic conditions. CpG oligonucleotides play a critical role in immunity via the augmentation of Th1 and suppression of Th2 responses. In the present study we aimed to estimate the effectiveness of intranasal administration of CpG ODN plus Chenopodium album allergen in allergic asthma compared with the administration of allergen alone and to find out how CpG ODN therapy is useful in the treatment of allergen induced asthma. BALB/c Mice were intraperitoneally and intranasally sensitized with allergenic extract precipitated on aluminum hydroxide. Therapy with CpG/Ag was performed intranasally. After antigenic challenge, a number of Immunologic variables such as serum IgE and IgG, systemic and local IL-10 and IFN-Gamma were studied in splenocytes, and lung tissue culture supernatants, respectively. Our study indicated that intranasal administration of CpG/Ag had significant increases in both systemic and local levels of IL-10 and IFN-Gamma [p

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

Interaction of CpG-oligodeoxynucleotides with toll like receptor 9 induces apoptosis and modulates metaloproteinase-2 activity in human intestinal epithelium

Recent reports have indicated different effects of immunostimulatory sequences containing CpG-Oligodeoxynucleotides [ODN] on various immune cells. However, the exact role of CpG-ODN in the human gut is unclear. In the present study, we assessed potential effects of CpG-ODN on non lymphoid cell [intestinal epithelial cell line HT-29] on a dose-response and time-course basis. Intestinal epithelial cell line HT-29 was treated with CpG-ODN [CpG 2006] and lipopolysaccharide [LPS] at 5, 10, 25, 50 micro g/ml and 1, 5, 10 micro g/ml concentrations, respectively. Following treatments, dose-response and time-course cytotoxicity using a colorimetric method, Metaloproteinase-2 [MMP-2] activity [using gelatin zymography] and apoptosis [using annexin-v flowcytometry method] assays were performed. Chloroquine treatment was also used for its inhibitory effect on endosomal acidification process to verify specific CpG-ODN and Toll Like Receptor 9 [TLR9] interactions. Cytotoxicity analysis of CpG-ODN showed that CpG-ODN increased significantly the proliferation of CpG-ODN treated cells, as compared to untreated cells, at concentrations of 10-25 micro g/ml [p<0.05]. Overall MMP-2 activity analysis showed significant differences between treated and untreated cells. However, minimal changes were observed when MMP-2 activity was assessed per cell. Moreover, CpG-ODN treated cells demonstrated an increasing apoptosis rate of 0.8%, 6.46% and 14.21% at concentrations of 5, 10, 25 micro g/ml, respectively. Collectively, our data indicated that intestinal epithelial cell line HT-29 is highly responsive to CpG effect in vitro and exhibits modified activities. The direct CpG-ODN and TLR-9 interactions in HT-29 cells could provide new approaches in malignant tumor therapeutic strategies

Dermal wound fibroblasts and matrix metaloproteinases [MMPs]: their possible role in allergic contact dermatitis

This study was conducted to examine if allergic contact dermatitis [ACD] alters the expression ofMMPs in human dermal fibroblasts. Fibroblasts are the primary source for MMP and matrix production in skin. MMPs are known to involve in a number of physiological and pathological processes. Some published data indicated a gelatinaselike activity in acute and chronic phases of allergic contact dermatitis. However, no exact source of gelatinase activity was demonstrated. Moreover, little is known about the role of MMPs in immune responses. To study and predict the pathophysiological effects of [MMP-2] in allergic contact dermatitic [ACD] patients, we established an in vitro tissue culture survey based on fibroblast explanted from ACD wounds and normal tissues respectively. We also employed a precise proliferation assay [i.e. MTT; 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide] to analyze and compare three ACD vs. three normal cell strains Parallel to MTT assay, we assessed the activity as well as the kinetics of gelatinase [MMP-2] in conditioned media using a zymogeraphy analysis. There was a significant difference in proliferation capacity between mean ACD fibroblast strains vs. mean normal cells, particularly in days 6 to 8 post explantation, 492.5 +/- 6.6 vs. 361.75 +/- 8.25 respectively. Zymoanalyses indicated significant differences betweenACD cells and normal fibroblasts both in time-course and MMP-2 activity per cell fashions, 163.7 +/- 16.21 for meanACD fibroblasts vs. 130 +/- 9.09 for normal cells respectively. These data suggest that fibroblasts overproliferated in the process ofACD. Moreover, simultaneous overexpression of MMPs observed inACD fibroblasts vs. normal strains, is indicative of altered fibroblast functionality in the process of allergic contact dermatitis. The activity per cell analysis showed that MMP-2 expression in ACD fibroblasts is independent of cell number, suggesting that either intra- or inter-cellular control signals are also altered and that ACD fibroblasts exhibit hyper-responsiveness to mitogenic or fibrogenic stimulants. Altogether, these data address the chronocity and non-healing tendency of ACD wounds. However, more studies are required to examine possible MMPs inhibition and differential expression of mytogenic, fibrogenic and antifibrogenic cytokines in ACD wound beds. In particular, MMP-2 is postulated to be an aim for further gene therapy protocols