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. 2022 May 18;18(5):e1010003. doi: 10.1371/journal.ppat.1010003

Fig 5. TLR3 dependent recognition of Cryptosporidium infection.

Fig 5

We sought to identify the pattern recognition receptor that is activated to induce IFNλ production in response to Cryptosporidium infection. (A) ELISA of HCT-8 cultures infected with C. parvum or treated with the indicated agonist for 24 hours to assess IFNλ production in response to stimulus against a variety of pattern recognition receptors, n = 3. IFNλ is induced in response to the following stimuli: infection, Poly(I:C), lipofected Poly(I:C), and ssPolyU RNA One-way ANOVA with Dunnett’s multiple comparisons *** p <0.001 **** p <0.0001. (B) Infection of wild type control mice (B6129) compared to mice lacking MAVS. Fecal luminescence measured every 2 days. 3–4 mice per group. Representative of 2 biological replicates (1.18 and 1.04-fold increase) n = 2 Standard t-test of area under the curve across all biological replicates. (C) Infection of wild type control mice (C57B6N/J) compared to mice lacking TLR7. Fecal luminescence measured every 2 days. 3–4 mice per group. Representative of 3 biological replicates (1.3, 0.70, 2.2-fold increase) n = 3 Standard t-test of area under the curve across all biological replicates. (D) Infection of wild type control mice (B6129) compared to mice lacking TLR3. Fecal luminescence measured every 2 days. An increase of 8-fold in oocyst shedding was observed. 3–4 mice per group. Representative of 3 biological replicates (8, 12.4, 3.8-fold respectively) n = 3 Standard t-test of area under the curve across all biological replicates * p <0.05. (E) IL-18 protein detected from ileal biopsies of infected mice wild type (B6129) compared to Tlr3-/- at day 2 post infection. Standard t-test *** p <0.001. 11 mice per group, n = 2. (F) IFNλ protein detected from ileal biopsies of infected mice wild type (B6129) compared to Tlr3-/- at day 2 post infection. Standard t-test *** p <0.001. 11 mice per group, n = 2. Dotted line represents the limit of detection.