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. 2015 Dec 31;9(12):e0004234. doi: 10.1371/journal.pntd.0004234

Fig 3. Oxidation of glycans results in a decreased production of IL-4 and IL-10 and an increase of IFNγ by parasite specific splenocytes.

Fig 3

BALB/c mice (n = 5 per group) were infected with 15 metacercariae. After three weeks, animals were bled, sacrificed and spleens and hepatic draining lymph nodes were removed. Splenocytes were cultured in the presence of 75 μg/mL of FhTE, FhCB (oxidation negative control) or FhmPox (oxidized FhTE). Culture supernatants were collected and analyzed by ELISA for IL-4, IL-5, IL-10 or IFNγ (A). Alternatively, IL-4, IL-10 and IFNγ were detected by quantitative RT-PCR. Results are shown as the ratio of mRNA amplification for a specific cytokine and the GAPDH (B). Also, cells from hepatic draining lymph nodes were stimulated with FhTE, FhCB or FhmPox (75 μg/mL) and IL-5 and IL-10 on the supernatants were determined by ELISA (C). Total antibodies in sera from infected and non-infected animals specific for FhTE or FhmPox were evaluated on plates coated with parasite components (10 μg/ml) and with serial dilution of animal sera (D). Results are expressed as the mean of three independent experiments (±SD, indicated by error bars). Asterisks indicate statistically significant differences (**p < 0.01; *p < 0.05) with respect to FhTE-stimulated splenocytes from non-infected animals.