Journal:

Article Title: Colonic epithelial cell mediated suppression of CD4 T cell activation

doi: 10.1136/gut.2003.029967

Figure Lengend Snippet: Colonic epithelial cells (CEC) inhibit the response of T cells to conventional mitogens. Splenic CD4 T cells (A, B, D, F, G) or colonic lamina propria CD4 T cells (C, E) were cultured with freshly isolated C57BL/6 CEC or macrophages (Mφ) in the presence of mitogenic CD3 antibody for 72 hours prior to flow cytometric analysis for expression of CD25 on splenic (A) and lamina propria T cells (C), CD45RB on splenic CD4 T cells (B), cell cycle progression of splenic T cells (D) and lamina propria T cells (E), and cytokine production of splenic T cells using cytokine bead assays (F, G). In (A), (B), and (C), the filled in and solid line profiles represent levels of antibody staining of T cells after coculture with Mφ and CEC, respectively, and the broken lines represent the isotype matched control antibodies. (D, E) Proportion of splenic and lamina propria CD4 T cells in the G1, S, and G2 phases of the cell cycle. (F, G) Cytokines produced by splenic T cells after culture with CEC and Mφ in the presence of mitogen, respectively. Results are representative of those obtained from at least six independent experiments. IFN-γ, interferon γ; IL, interleukin; TNF-α, tumour necrosis factor α.

Article Snippet: Cytokine multiplex bead assay kits (Becton Dickinson) were used to measure levels of interferon γ (IFN-γ), tumour necrosis factor α (TNF-α), interleukin (IL)-5, IL-4, and IL-10 in CEC:T and Mφ:T cell conditioned media.

Techniques: Cell Culture, Isolation, Expressing, Staining, Control, Produced

Journal:

Article Title: Colonic epithelial cell mediated suppression of CD4 T cell activation

doi: 10.1136/gut.2003.029967

Figure Lengend Snippet: Responses of antigen specific T cells are inhibited by colonic epithelial cells (CEC). I-Ad-restricted DO11.10 CD4 T cells were cocultured with OVA primed DO11.10 CEC or macrophages (Mφ) in the presence or absence of OVA for 72 hours prior to flow cytometric analysis of T cells for expression of CD25 (A), CD45RB (B), cell cycle progression (C), and cytokine production (D). (A, B) Cells cultured in the absence or presence of OVA: **p<0.005 (A) and **p<0.005 (B) comparing CD4 T cell activation in the presence of BALB/C (I-Ad) CEC or Mφ. (C) Proportion of CD4 T cells in the G1, S, and G2 phases of the cell cycle, respectively, with the plus and minus symbols referring to the presence or absence of OVA in the coculture. (D) Cytokine production by CD4 T cells cocultured with CEC and Mφ, respectively. Results are representative of those obtained from at least eight independent experiments. IFN-γ, interferon γ; IL, interleukin; TNF-α, tumour necrosis factor α. (E) C57BL/6 (I-Ab) CEC can present OVA to an OVA specific I-Ab restricted CD4 T cell hybridoma KZO. Cultured CEC were pulsed with OVA, washed, and incubated for 24 hours with KZO, which had been transfected with an NFAT-lacZ reporter gene construct. T cell hybridoma activation and lacZ activity were determined spectrophotometrically using the chromogenic substrate and measuring absorbance at 595 nm (OD 595).

Article Snippet: Cytokine multiplex bead assay kits (Becton Dickinson) were used to measure levels of interferon γ (IFN-γ), tumour necrosis factor α (TNF-α), interleukin (IL)-5, IL-4, and IL-10 in CEC:T and Mφ:T cell conditioned media.

Techniques: Expressing, Cell Culture, Activation Assay, Incubation, Transfection, Construct, Activity Assay