Phosphorylated dextran (P-Dex) is an acidic polysaccharide that functions as an immune adjuvant. P-Dex is known to regulate immune response by maintaining a balance between Th1 and Th2 cells in vitro, and thus may also be important in the control of allergic reactions. In the current study, we report the optimum conditions required for the efficient phosphorylation of dextran without toxicity. We found that when dextran was heated at 160${^{\circ}C}$ for 24 h in phosphate buffer (pH 5.0), the resulting P-Dex demonstrated the highest phosphorus content (6.8%). We also report that P-Dex enhances mitogenic activity in mouse splenocytes and induces expression of CD69 and CD86 on the surface of B cells and dendritic cells (DC) in vitro. Oral administration of P-Dex to ovalubmin (OVA)-immunized mice was found to reduce antigen-induced cell proliferation and suppress the expression of CD86 on Th2-inducing DC via exogenous OVA stimulation. P-Dex was also found to increase IL-10 expression in the splenocytes of treated mice. These findings suggest that oral administration of P-Dex increases immunological tolerance and improves the specificity of immunological response to specific antigens.
The distributions and morphological characteristics of neurons displaying immunoreactivity to the catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-${\beta}$-hydroxylase(DBH), and phenylethanolamine-N-methyltransferase (PNMT) were examined in the adjacent sections of the olfactory bulb of the Striped Field Mouse(Apodemus agrarius coreae). None of these cell groups displayed either DBH or PNMT immunoreactivity. Many TH-immunoreactive neurons were present in the olfactory bulb. The vast majority of such cells occurred in the glomerular layer as periglomerular cells surrounding the glomeruli. Numerous addtional cells were present in the external plexiform layer, and scattered in the mitral cell layer and internal plexiform layer. Also TH-immunoreactive neurons were found in the glomerular layer and granular layer of the accessory olfactory bulb.
In this study, the pathway and date of migrating Primordial germ cells (PGCs) were observed light microscopically and ultrastructural changes of them during migration were observed by electron microscopic examination. For these purpose, alkaline phosphatase reactions were used for identifying the PGCs and acid phosphatase reactions were used for observing their degenerating activities. Also, effects of actinomycin D on the migration of PGCs were examined. According to these results, at the 9th gestation day, PGCs were observed in the endodermal cells of yolk sac, at the 11th gestation day, they were seen in the hindgut and then entered into the dorsal mesentery by the 13th gestation day. At the 14th gestation day, they were located in the genital ridges. When PGCs were located in the hindgut and genital ridges, the positive reactions of alkaline phosphatase were dominated, but acid phosphatase reactions were limited in all stage except they were in dorsal mesentery. However, these reactions were lessened in case of actinomycin D treatment. By electron microscopic examination, PGCs had pseudopodia, tail process, trailing cytoplasm and nuage as the ultrastructural characteristics. In addition, these morphological features were damaged by actinomycin D treatment.
In order to define the morphological changes of the secreting cells of prothoracic gland during larva-pupal molt, ultrastructural observations were carried out using Bombyx mori L. as the experimental material. At first stage of present experiment, 4 day old 5th instar larva, the polyhedral secreting cells were centrally located in the prothoracic gland surrounded by the connective sheath. The secreting cells were attached to the neighboring cells by the prominent desmosomes, and the plasma membrane contacted with connective sheath were highly infolded. In cytoplasm, the most of the cell organelles, such as rod-like mitochondria, rough surfaced endoplasmic reticulum, ribosome were developed. As the stages advance from larva to pupa, general feature of the secreting cells were retained, but structural changes of the various cytoplasmic organelles-ribosome, rough surfaced endoplasmic reticulum, mitochondria, Golgi apparatus, lamellar body, and vesicle-were noted. In the perinuclear cytoplasm of the secreting cells at the stage of 6 day old 5th instar larva, it is peculiar that only a large amount of ribosomes were distributed and the other organelles were retreated from the juxtanuclear region. Just before and after spining cocoon, these features were more remarkable. Rough surfaced endoplasmic reticulum were gradually increased from the stage just before spining cocoon to the pharate pupa. Rod-like mitochondria with irregular cristae and the matrix showing low density were distributed throughout the cytoplasm in the secreting cells of the 4 day old 5th instar larva. Sometimes, longitudinally distended and curved mitochondria were observed. At the stage of pharate pupa, most of mitochondria were deformed. The rod-like mitochondria of the secreting cells of pupal prothoracic gland were narrower than those of 4 day old 5th instar larva, and the electron density of the mitochondrial matrix is increased in pupa. Golgi apparatus were a few in number in both stages, last instar larva and spining cocoon. In stages of the pharate pupa, the Golgi apparatus were frequently observed. Cytoplasmic vesicles were observed for the first time in the secreting cells of one day after spining cocoon, and the number and the size of cytoplasmic vesicles were distinctly increased inpharate pupa and just after pupation. In the secretory cells of the PG, it in concluded that the RER was closely related to syntheting the enzymes seem to produce the ecdysone.
Antigen-specific T cell clones were obtained from mice immunized with Fusobacterium nucleatum ATCC 10953(F .nucleatum) and/or Porphyromonas gingi valis 381(P. gingivalis). 10 Balb/c mice per group were immunized with F. nucleatum followed by P. gingivalis, or with P. gingivalis alone by intraperitoneal injection of viable microorganisms. Spleen T cells were isolated and stimulated in vitro with viable P. gingivalis cells to establish P. gingivalisspecific T cell clones. T cell phenotypes and cytokine profiles were determined along with T cell responsiveness to F .nucleatum or P. gingivalis. Serum IgG antibody titers to F. nucleatum or P. gingivalis were also determined by ELISA. All the T cell clones derived from mice immunized with F. nucleatum followed by P. gingivalis demonstrated Th2 subsets, while those from mice immunized with P. gingivalis alone demonstrated Th1 subsets based on the flow cytometric analysis and cytokine profiles, All T cells clones from both groups were cross-reactive to both P. gingivalis and F. nucleatum antigens. Phenotypes of T cell clones were all positive for CD4. Mean post-immune serum IgG antibody levels to F. nucleatum or P . gingivalis were significantly higher than the pre-immune levels(p <0.01, respectively). There were no significant differences in the antibody titers between the two groups. It was concluded that P. gingivalis-specific T cells initially primed by cross-reactive F. nucleatum antigens were polarized to Th2 subsets, while T cells stimulated with P. gingivalis alone maintained the profile of Th1 subset.
Objective: To explore the immune reconstitution of $CD4^+T$ cells after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and its relationship with invasive fungal infection (IFI) in patients with hematological malignancies. Materials and Methods: Forty-seven patients with hematological malignancies undergoing Allo-HSCT in Binzhou Medical University Hospital from February, 2010 to October, 2014 were selected. At 1, 2 and 3 months after transplantation, the immune subpopulations and concentration of cytokines were assessed respectively using flow cytometry (FCM) and enzyme linked immunosorbent assay (ELISA). The incidence of IFI after transplantation and its correlation with immune reconstitution of $CD4^+T$ cells were investigated. Results: The number of $CD4^+T$ cells and immune subpopulations increased progressively after transplantation as time went on, but the subpopulation cell count 3 months after transplantation was still significantly lower than in the control group (p<0.01). In comparison to the control group, the levels of interleukin-6 (IL-6) and IL-10 after transplantation rose evidently (p<0.01), while that of transforming growth factor-${beta}$ (TGF-${beta}$) was decreased (p<0.01). There was no statistically significant difference level of interferon-${\gamma}$ (IFN-${\gamma}$) (p>0.05). The incidence of IFI was 19.2% (9/47), and multivariate logistic regression revealed that IFI might be related to Th17 cell count (p<0.05), instead of Th1, Th2 and Treg cell counts as well as IL-6, IL-10, TGF-${beta}$ and IFN-${\gamma}$ levels (p>0.05). Conclusions: After Allo-HSCT, the immune reconstitution of $CD4^+T$ cells is delayed and Th17 cell count decreases obviously, which may be related to occurrence of IFI.
Allergic asthma is one of the most enduring diseases of the airway. The T-helper cells and regulatory T-cells are critically involved in inflammatory responses, mucus hypersecretion, airway remodelling and in airway hyper-responsiveness. Cigarette smoke (CS) has been found to aggravate inflammatory responses in asthma. Though currently employed drugs are effective, associated side effects demand identification and development of novel drugs with negligible or no adverse effects. Rutin, plant-derived flavonoid has been found to possess antioxidant and anti-inflammatory effects. We investigated the ability of rutin to modulate T-cells and inhibit inflammation in experimentally-induced asthma in cigarette smoke exposed mice. Separate groups of neonatal mice were exposed to CS for 10 days from post-natal days 2 to 11. After 2 weeks, the mice were sensitized and challenged with ovalbumin (OVA). Treatment group were given rutin (37.5 or 75 mg/kg body weight) during OVA sensitization and challenge. Rutin treatment was found to significantly inhibit cellular infiltration in the airways and Th2 and Th17 cytokine levels as well. Flow cytometry revealed effectively raised $CD4^+CD25^+Fox3^+$ Treg cells and supressed Th17 cell population on rutin treatment. Airway hyper-responsiveness observed following CS and OVA challenge were inhibited by rutin. $NF-{\kappa}B$ and iNOS, chief regulators of inflammatory responses robustly activated by CS and OVA were down-regulated by rutin. Rutin also inhibited the expression of matrix metalloproteinase 9, thereby aiding in prevention of airway remodelling in asthma thereby revealing to be a potent candidate in asthma therapy.
In Trichinella spiralis infection, type 2 helper T (Th2) cell-related and regulatory T ($T_{reg}$) cell-related immune responses are the most important immune events. In order to clarify which Toll-like receptors (TLRs) are closely associated with these responses, we analyzed the expression of mouse TLR genes in the small intestine and muscle tissue during T. spiralis infection. In addition, the expression of several chemokine- and cytokine-encoding genes, which are related to Th2 and $T_{reg}$ cell mediated immune responses, were analyzed in mouse embryonic fibroblasts (MEFs) isolated from myeloid differentiation factor 88 (MyD88)/TIR-associated proteins (TIRAP) and Toll receptor-associated activator of interferons (TRIF) adapter protein deficient and wild type (WT) mice. The results showed significantly increased TLR4 and TLR9 gene expression in the small intestine after 2 weeks of T. spiralis infection. In the muscle, TLR1, TLR2, TLR5, and TLR9 gene expression significantly increased after 4 weeks of infection. Only the expression of the TLR4 and TLR9 genes was significantly elevated in WT MEF cells after treatment with excretory-secretory (ES) proteins. Gene expression for Th2 chemokine genes were highly enhanced by ES proteins in WT MEF cells, while this elevation was slightly reduced in MyD88/$TIRAP^{-/-}$ MEF cells, and quite substantially decreased in $TRIF^{-/-}$ MEF cells. In contrast, IL-10 and $TGF-{\beta}$ expression levels were not elevated in MyD88/$TIRAP^{-/-}$ MEF cells. In conclusion, we suggest that TLR4 and TLR9 might be closely linked to Th2 cell and $T_{reg}$ cell mediated immune responses, although additional data are needed to convincingly prove this observation.
Objectives : Asthma is a chronic inflammatory disorder of the airways by many cells such as mast cells, Th2 lymphocytes and eosinophile. The present study was aimed to evaluate the effects of Salviae miltiorrhizae (SM) on T cell cytokine production, mast cells. and eosinophils, Methods : We screened 13 herbs to find compounds with potential to control Th cytokine production. using concanavalin A (con A)-activated splenocyte cultures. Con A-activated $IFN-\gamma$ and IL-4 levels in supernatants of splenocyte cultures. Bone marrow derived mast cells (BMMC) were incubated with SM and then the expressions of membrane proteins of BMMC were analyzed by fluorescence activated cell sorter (FACS). BALB/c mice sensitized to ovalbumin (OVA) were challenged with aerosolized OVA for 6 weeks. During the last weeks some mice were treated with SM. Then eosinophils in bronchoalveolar lavage fluid (BALf) were counted and pathologic changes of lung tissue were observed with hematoxylin-eosin stain. Results : SM increased $IFN-\gamma$ level on splenocyte culture significantly. but had no significant effects on expressions of ICAM-1, CD62L, integrin $a_4$. c-kit, IL-3 receptors. CD11a, or IgE receptors of BMMC. SM treatment significantly inhibited eosinophil infiltrates in BALf and peribronchial lung inflammation. Conculusions : The present data suggested that SM may have an effect on Th cytokine secretion and eosinophils associated with asthma responses. Therefore SM might be of therapeutic value in treating asthma.
Kim, Tae-Gyun;Park, Sung-Min;Kang, Hee;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
Journal of Physiology & Pathology in Korean Medicine
/
v.22
no.4
/
pp.754-761
/
2008
The purpose of this study was to evaluate the effect of Baekhasuoyijung-Tang(BHSYJT)on mouse T cell cytokines. The proliferation of mouse CD4 T cells under the influence of BHSYJT extract was measured. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 in various concentrations of BHSYJT extract, it increased proliferation of CD4 cells by 28% in $10{\mu}g/m{\ell}$ concentration and by 32% in $100{\mu}g/m{\ell}$ concentration. Treatment of CD4+ T cells stimulated by anti-CD3e and anti-CD28 with BHSYJT resulted in reduction of $IFN-{\gamma}$,but IL-4 levels is not changed. Oral administration of BHSYJT resulted in increase of both CD4+ and CD8+ T cell population in Balb/c mice by 11%. Oral administration of BHSYJT resulted in reduction of serum $IFN-{\gamma}$ level by 27% but, IL-4 level is not changed. CD4+ T cells under Th1/Th2 polarizing conditions for 3 days with BHSYJT resulted in decrease of $IFN-{\gamma}$ level in TH1 cells. Experimental results of this study show that BHSYJT helps to reduce secretion of $IFN-{\gamma}$ by mouse T helper cell in vitro and it had the same effect in vivo. Thus, it can be concluded that use of BHSYJT is an effective treatment for correcting immune imbalance in immune disorders and autoimmune diseases by reducing secretion of cytokine by Th1 cells.
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