Abstract
Chondroitin sulfate (CS) was administered orally to BALB/c mice immunized intraperitoneally with ovalbumin (OVA) and/or dinitrophenylated OVA. The titers of antigen-specific IgE and IgG1 in mouse sera were determined. The antigen-specific IgE production by mice fed ad libitum with CS was significantly inhibited. We also examined the effect of feeding CS on immediate-type hypersensitivity. One hour after antigen stimulation, the ears of mice fed with CS swelled less than those of the control mice. Furthermore, the rise in serum histamine in the mice fed with CS under active systemic anaphylaxis was significantly lower than that in the controls. We next examined the pattern of cytokine production by splenocytes from mice followed by re-stimulation with OVA in vitro. The splenocytes from the mice fed with CS produced less interleukin (IL)-5, IL-10, and IL-13 than those from the control group. In contrast, the production of interferon-gamma and IL-2 by the splenocytes of mice fed with CS was not significantly different from those in the control mice. In addition, the production of transforming growth factor-beta from the splenocytes of mice fed with CS was significantly higher than that of the control mice. Furthermore, we showed that the percentages of CD4(+) cells, CD8(+) cells, and CD4(+)CD25(+) cells in the splenocytes of mice fed with CS are significantly higher than those of the control. These findings suggest that oral intake of CS inhibits the specific IgE production and antigen-induced anaphylactic response by up-regulating regulatory T-cell differentiation, followed by down-regulating the Th2 response.
Highlights
To further assess the involvement of the immunological process of Chondroitin sulfate (CS) intake, we analyzed the differentiation in splenocytes, Peyer’s patch (PP) cells, mesenteric lymph node (MLN) cells, and intestinal intraepithelial lymphocytes (IELs) using flow cytometry (FCM)
The percentage of CD4ϩ T cells in the splenocytes from the mice fed with CS (27.7%)
We first showed that feeding CS to mice inhibited specific IgE and IgG1 production in response to DNP-OVA in vivo, centage of CD8ϩ T cells in the splenocytes from the mice fed but DNP-specific IgG1 production was not significantly with CS (11.8%) was significantly higher than that of the affected
Summary
Animals and Administration Protocols—Inbred specific pathogen-free BALB/c mice (female, 6 weeks of age) were purchased from Charles River Japan (Yokohama, Japan). Induction of Active Systemic Anaphylactic Shock and Measurement of Histamine in Plasma—The mice immunized with OVA were challenged intraperitoneally with 0.2 ml of phosphate-buffered saline (PBS) containing OVA (1 mg/mouse) to induce anaphylactic shock. FCM Analysis of Splenocytes—The collected splenocytes from mice were washed twice in PBS containing 2% FBS, and cell staining analysis was carried out in the following way [28, 34]. 100 l of splenocytes at 2.0 ϫ 106 cells/ml in PBS containing 2% FBS and 0.1% NaN3 was mixed with 1 l of anti-CD16/CD32 monoclonal antibody (BD Pharmingen, San Diego, CA) in a tube and reacted for 5 min at 4 °C. A single cell suspension of lymphocytes in BD Pharmingen Stain Buffer containing 2% FBS was incubated with 50 l of properly diluted monoclonal antibody at 4 °C for 30 min.
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