Helminth Infection Enhances Disease in a Murine TH2 Model of Colitis Meaghan M. Hunter, Arthur Wang, Derek M. Mckay  Gastroenterology  Volume 132, Issue 4, Pages 1320-1330 (April 2007) DOI: 10.1053/j.gastro.2007.01.038 Copyright © 2007 AGA Institute Terms and Conditions

Figure 1 Infection with H diminuta increases the severity of oxazolone-induced colitis as assessed by (A) change in body weight, (B) clinical scores, and (C) shortening of the colon. Mice were infected with H diminuta 8 days prior to oxazolone treatment (3 mg/100 μL of 50% EtOH IR) and autopsied 3 days postoxazolone (mean ± SEM; n = 16–20 mice from 4–5 separate experiments; *P < .05 compared with control; †P < .05 compared with oxazolone only). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 2 Infection with H diminuta prior to oxazolone (OX; 3 mg/100 μL of 50% EtOH IR) enhances colonic inflammation. Representative photomicrographs show normal colonic architecture in naive control mice and those infected with H diminuta. Oxazolone treatment results in loss of colonic architecture, edema, goblet cell depletion, and a mixed immune cell infiltration (3 days posttreatment) that has resolved to a large extent by 7 days posttreatment. Colonic sections from H diminuta + oxazolone-treated mice show more severe inflammation that is not resolved 7 days after oxazolone, in which eosinophils (inset) are prominent. Depletion of NK and NK-T cells with an anti-NK1.1 AB prevents oxazolone-induced histopathology and is partially effective in blocking the effect of H diminuta + oxazolone (original magnifications are given; images are representative of 4–8 mice/group). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 3 Infection with H diminuta prior to oxazolone (OX; 3 mg/100 μL of 50% EtOH IR) enhances colonic inflammation as assessed by (A) histology damage scores and (B) MPO activity (mean ± SEM; n = 16–20 mice from 4–5 separate experiments; *P < .05 compared with control; †P < .05 compared with oxazolone). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 4 (A) Analysis of colonic eosinophil peroxidase (EPO) activity in colonic extracts reveals increases in H diminuta (H.d) and oxazolone (OX; 3 mg IR)-only treated mice and further enhancement in the cotreatment group examined 3 days postoxazolone. Depletion of NK and NK-T cells via anti-NK1.1 AB completely blocks OX-induced increases in EPO. Mice examined 7 days post-OX still had slightly elevated EPO activity, and again, this was on average greater in H.d + OX-treated mice (mean ± SEM; n = 9–13 mice from 3–4 experiments for the 3-day treatment and n = 3–4 mice for the 7-day experiment; *P < .05 compared with control [con]; †P < .05 compared with oxazolone). (B) Eosinophils numbers/section in the various experimental groups (mean ± SEM; n = 3–4; data for 7-day H diminuta + oxazolone are observations from tissue obtained from mice humanely sacrificed on days 5–7 postoxazolone). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 5 In vitro conA (2 μg/mL; 48 hours) challenge of spleen and MLN cells results in increased IL-4, -5, -10, and -13 production from cells isolated from H diminuta-infected mice ± oxazolone. (E) TGFβ production by stimulated immune cells (n = 4) (mice received 5 H diminuta cysticercoids 8 days before oxazolone [3 mg, IR] with autopsy at 72-hour postoxazolone; mean ± SEM, n = 8–9 mice from 3 separate experiments for spleen cells, n = 6–8 mice from 2 separate experiments for MLN cells; *P < .05 compared with control; †P < .05 compared with oxazolone; ∞P < .05 compared with H diminuta). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 6 (A) Representative RT-PCR products for eotaxin, IL-5, TGFβ, and β-actin mRNA expression in extracts of distal colon from the various treatment groups. (B) The densitometric analysis of the RT-PCR products (n = 4) compared with β-actin, where oxazolone ± H diminuta results in increased IL-5 and eotoxin expression, and subtler increases in TGFβ mRNA (mean ± SEM; *P < .05 compared with control; †P < .05 compared with oxazolone; ∞P < .05 compared with H diminuta). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 7 Infection with H diminuta hindered recovery from oxazolone-induced colitis (3 mg, IR) as assessed by (A) animal morbidity, (B) cytokine production from conA (2 μg/mL) stimulated splenocytes (n = 4 for control and oxazolone, and n = 1, 2, and 1 for 4, 5, and 7 days post-H diminuta + oxazolone, respectively [ie, mice humanely sacrificed matching A]; *P < .05 compared with control; statistics not done for H diminuta cotreated mice; mean ± SEM; ng/mL of cytokine; nd, not done). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions

Figure 8 In vivo depletion of NK and NK-T cells via intraperitoneal delivery of an anti-NK1.1 AB (20 μg/mouse on days 1, 2, 5, 8, and 11) reduces the pro-colitic effect of oxazolone (OX; 3 mg IR, assessed 72 hours later) and that induced by the combined treatment of H diminuta + oxazolone as assessed by (A) clinical disease scores, (B) colon lengths, and (C) histology damage scores, and (D) MPO activity levels (mean ± SEM, with n = 4 mice; a, b, and c indicate statistically significantly different groups [P < .05]; Ig, isotype-matched irrelevant antibody). Gastroenterology 2007 132, 1320-1330DOI: (10.1053/j.gastro.2007.01.038) Copyright © 2007 AGA Institute Terms and Conditions