Journal: International Journal for Parasitology

Article Title: The helminth product, ES-62, protects against airway inflammation by resetting the Th cell phenotype

doi: 10.1016/j.ijpara.2012.12.001

Figure Lengend Snippet: Exposure to the phosphorylcholine-containing glycoprotein, ES-62, in vivo does not modulate ovalbumin (OVA)-induced elevation of IgG1 and IgG2a but inhibits IgE and suppresses Th2 cells. Serum samples from each mouse were obtained on day 28. Samples were analysed by ELISA for OVA-specific IgE (A) and total IgE (B), at 1/80 and 1/200 dilutions, respectively, and OVA-specific IgG1 (C) and IgG2a (D) over the full titration range where data for each group are presented as the mean ± S.E.M. ( n = 6 mice/group) and ∗∗∗ P ⩽ 0.001, OVA + ES-62 compared with the OVA group. The data are from one experiment representative of two. In E, draining lymph nodes (DLN) cells from each of the four treatment groups were stained for expression of IgE. (F) DLN sections from OVA and OVA + ES-62 groups were stained for B220 + B cells, CD3 + T cells and for the indicated transcription factor (either GATA3, Foxp3 or Tbet) and the transcription factor positive cells were quantified within the T cell regions. Exemplar tissue maps from a single experiment of DLNs from OVA and OVA + ES-62 groups displaying the relevant distribution of such transcription factor positive cells (red) within the T cell paracortical region (white) surrounded by B220 + B cells (blue) and their quantitation of numbers of the relevant transcription factor positive T cells by laser scanning cytometry (LSC) are shown, together with an exemplar relocated image of a portion of the section from the OVA group where Foxp3 is stained in red and CD3, green (F). In an independent experiment, the proportions of Foxp3-expressing unstimulated CD4 + T cells from the DLN of pooled mice from the four treatment groups are shown, with the percentage of cells in the Foxp3 + CD4 + quadrant annotated to the right of the plots (G). Pooled DLN cells from the four treatment groups were cultured with medium (I) or phorbol 12-myristate 13-acetate (PMA) plus ionomycin (IONO) (H, J) in the presence of Brefeldin A before staining for intracellular expression of IL-10 (H) or IFNγ (I, J) prior to FACS analysis: cytokine expression ( y -axis) was plotted against side scatter ( x -axis) and the percentage of DLN cells in the cytokine positive gate is shown and annotated to the right of the plots. (J) The Mean Fluorescence Intensity (MFI) values for IFNγ production for the four treatment groups are PBS: 24,701; ES-62: 21,915; OVA: 23,439 and OVA + ES-62: 27,596. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Total IgE and OVA-specific IgG1, IgG2a and IgE levels were measured in serum by ELISA (BD Biosciences, UK) following the manufacturer’s instructions using Maxisorp ELISA plates (Nunc, Roskilde, Denmark).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Titration, Staining, Expressing, Quantitation Assay, Cytometry, Cell Culture, Fluorescence