PMC

PI and TPx induce the secretion of anti-inflammatory agents from RAW macrophages. The level of PGE₂, IL-10, and IL-12 in the culture supernatants of PI- and TPx-activated macrophages was measured by enzyme-linked immunosorbent assay and compared to cells incubated with PBS alone. The data represents the mean of four experiments ± the SE. ✽✽✽, P < 0.001.

TPx induces cultured RAW macrophages to an alternative activated phenotype. After treatment of RAW macrophages with purified recombinant TPx, the expression of Arg1 (a) and iNOS (b) was measured by RT-PCR (the data are representative of four independent experiments). (c and d) Level of arginase activity and NO, respectively, measured in the culture supernatants of the same cell preparations. The data are presented as the mean ± the SE. ✽, P < 0.05; ✽✽, P < 0.01; ✽✽✽, P < 0.001 (with respect to control treatments, either PBS or LPS).

Alternative activation of macrophages in BALB/c mice after an F. hepatica infection. Expression levels of β-actin, Fizz1, Arg1, Ym1, and iNOS were analyzed by RT-PCR in adherent peritoneal exudate cells isolated from mice prior to infection (day 0) and at 1, 7, 14, and 21 days postinfection (d.p.i.). The data shown are from a single mouse and are representative of the findings from four mice examined at each time point (the expression of each gene was similar in each mouse).

The PI fraction of F. hepatica ES products contains TPx and induces Th2 responses when intraperitoneally injected into BALB/c mice. (a) Antigen-specific production of IL-4 and IL-5 from spleen cells of BALB/c mice injected with ES products or fractions of ES products, termed PI and PII. Spleen cell cultures were stimulated with 20 μg of the relevant injected antigen—ES product, PI, or PII—per ml and PMA-anti-CD3 as a positive control. The data are presented as mean ± the standard error (SE) for four mice. (b) ES product, PI, and PII separated on a SDS-12.5% gel stained with Coomassie blue. (c) ES product, PI, and PII fractions separated by SDS-PAGE and analyzed by Western blots probed with antisera prepared against recombinant TPx.

Injection of PI and purified recombinant TPx induces the recruitment of alternatively activated macrophages into the peritoneum of BALB/c mice. (a) Macrophages recovered from the peritoneum of mice after an intraperitoneal delivery of PI or recombinant TPx exhibit increased expression of gene markers of alternative activation, including Fizz1 and Arg1, although Ym1 is not expressed as assessed by RT-PCR. The cells do not express iNOS, a marker of classically activated macrophages. (b and c) PI and TPx also induce the expression of Th2-associated cytokines IL-10 and TGF-β (b) but not the Th1 cytokines IL-12p40 and IL-18 (c). Agarose gels are representative of results obtained from four animals, and mean data obtained from densitometric analysis of all experiments are shown for IL-10 and TGF-β. Values for mRNA are expressed in relative absorbance units and are standardized per unit of β-actin per sample. ✽, P < 0.05 (for TPx compared to treatment with PBS).