Figure 2.

The reduced worm burden in trickle-infected C57Bl/6 mice is associated with increased tuft cell numbers. 6-8 week old C57Bl/6 mice were infected with 200 H polygyrus larvae according to the bolus and trickle infection regimes. (A) Paraffin embedded swiss rolls were stained with anti-DCLK1. The number of tuft cells were counted in 5 intact villi within the small intestine. Photographs from ten different areas of the small intestine were counted per animal (a minimum of five from areas with no granulomas and a minimum of three from areas with granulomas - Gr). (B) Mice were fasted for 6 hours followed by Evans Blue administration. Time from dye administration to the passing of dyed fecal pellets was measured and normalised to control animals. (C) Small intestines were dissected and scraped using a glass slide leaving only the serosa. The intestinal scrapings were weighed for each mouse and normalised to control animals. (D) Paraffin embedded swiss rolls were stained with Alcian blue. The average number of goblet cells in 5 intact and continuous villi within the small intestine was collected and normalised to the epithelial cell number. (E) RNA was extracted from whole intestinal tissue and Muc2 and Muc5a expression were measured normalised to the β-actin housekeeping gene. (F-J) IL-4, IL-13, RELMα, total IgA and IL-21 were measured in the intestinal scrapings by ELISA. Naive mice (grey circles) and mice infected according to the bolus (black circles) and trickle (white circles) regimes. Graphs represent pooled data from 2 experiments, bars represent the median, with a minimum of 2 mice per group per experiment. A normality test was performed (Anderson-Darling) followed by Kruskal Wallis tests with Dunn’s multiple comparisons test to test for statistical significance between trickle and bolus groups, n.s., not significant, *p<0.05, **p<0.01, ***p<0.001.