Epithelial cell–initiated inflammation plays a crucial role in early tissue damage in amebic infection of human intestine  Karl B. Seydel*,‡, Ellen Li*,§, Zhi Zhang*, Samuel L. Stanley  Gastroenterology  Volume 115, Issue 6, Pages 1446-1453 (December 1998) DOI: 10.1016/S0016-5085(98)70023-X Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 1 Inflammatory response of human intestinal xenografts to E. histolytica is inhibited by pretreatment with an intraluminal dose of an antisense oligonucleotide to the p65 subunit of human NF-κB. The mean quantity of human (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in human intestinal xenografts infected for 24 hours with E. histolytica (left panel of each graph) or sham infected with media alone (right panel of each graph), after intraluminal administration of either the nonsense oligonucleotide (n = 18), antisense oligonucleotide (n = 19), or no treatment (n = 10) is shown. The mean quantity of human IL-1β and IL-8 in E. histolytica–infected intestinal xenografts was significantly lower (P < 0.05) in antisense oligonucleotide–treated grafts than the levels in either nonsense oligonucleotide–treated grafts or untreated grafts. E. histolytica–infected grafts pretreated with the antisense oligonucleotide to the p65 subunit of human NF-κB (n = 16) also had significantly lower (P < 0.01) levels of MPO than E. histolytica–infected xenografts pretreated with the nonsense oligonucleotide (n = 16) or untreated grafts (n = 10). The levels of (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in uninfected xenografts, did not differ between nonsense oligonucleotide–treated (n = 10), antisense oligonucleotide–treated (n = 10), and untreated (n = 15) grafts. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 1 Inflammatory response of human intestinal xenografts to E. histolytica is inhibited by pretreatment with an intraluminal dose of an antisense oligonucleotide to the p65 subunit of human NF-κB. The mean quantity of human (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in human intestinal xenografts infected for 24 hours with E. histolytica (left panel of each graph) or sham infected with media alone (right panel of each graph), after intraluminal administration of either the nonsense oligonucleotide (n = 18), antisense oligonucleotide (n = 19), or no treatment (n = 10) is shown. The mean quantity of human IL-1β and IL-8 in E. histolytica–infected intestinal xenografts was significantly lower (P < 0.05) in antisense oligonucleotide–treated grafts than the levels in either nonsense oligonucleotide–treated grafts or untreated grafts. E. histolytica–infected grafts pretreated with the antisense oligonucleotide to the p65 subunit of human NF-κB (n = 16) also had significantly lower (P < 0.01) levels of MPO than E. histolytica–infected xenografts pretreated with the nonsense oligonucleotide (n = 16) or untreated grafts (n = 10). The levels of (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in uninfected xenografts, did not differ between nonsense oligonucleotide–treated (n = 10), antisense oligonucleotide–treated (n = 10), and untreated (n = 15) grafts. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 1 Inflammatory response of human intestinal xenografts to E. histolytica is inhibited by pretreatment with an intraluminal dose of an antisense oligonucleotide to the p65 subunit of human NF-κB. The mean quantity of human (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in human intestinal xenografts infected for 24 hours with E. histolytica (left panel of each graph) or sham infected with media alone (right panel of each graph), after intraluminal administration of either the nonsense oligonucleotide (n = 18), antisense oligonucleotide (n = 19), or no treatment (n = 10) is shown. The mean quantity of human IL-1β and IL-8 in E. histolytica–infected intestinal xenografts was significantly lower (P < 0.05) in antisense oligonucleotide–treated grafts than the levels in either nonsense oligonucleotide–treated grafts or untreated grafts. E. histolytica–infected grafts pretreated with the antisense oligonucleotide to the p65 subunit of human NF-κB (n = 16) also had significantly lower (P < 0.01) levels of MPO than E. histolytica–infected xenografts pretreated with the nonsense oligonucleotide (n = 16) or untreated grafts (n = 10). The levels of (A) IL-1β, (B) IL-8, and (C) neutrophils, measured by MPO assay in uninfected xenografts, did not differ between nonsense oligonucleotide–treated (n = 10), antisense oligonucleotide–treated (n = 10), and untreated (n = 15) grafts. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 2 Photomicrographs of H&E-stained sections from human intestinal xenografts infected with E. histolytica that were pretreated with intraluminal administration of either (A) antisense or (B) nonsense oligonucleotides to the p65 subunit of human NF-κB. An abundant influx of inflammatory cells can be seen in the lamina propria and mucosal tissue, as well as in the lumen of the human intestinal xenograft treated with nonsense oligonucleotide. In contrast, there is a marked reduction in the cellular response in the antisense-treated human intestinal xenograft, and E. histolytica trophozoites (arrowheads) can be seen in the lamina propria with minimal inflammation. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 2 Photomicrographs of H&E-stained sections from human intestinal xenografts infected with E. histolytica that were pretreated with intraluminal administration of either (A) antisense or (B) nonsense oligonucleotides to the p65 subunit of human NF-κB. An abundant influx of inflammatory cells can be seen in the lamina propria and mucosal tissue, as well as in the lumen of the human intestinal xenograft treated with nonsense oligonucleotide. In contrast, there is a marked reduction in the cellular response in the antisense-treated human intestinal xenograft, and E. histolytica trophozoites (arrowheads) can be seen in the lamina propria with minimal inflammation. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Permeability of human intestinal xenografts to fluorescently labeled dextran increases on infection with E. histolytica. The concentration of FITC-labeled dextran observed in the serum at various time points after injection of FITC-dextran into the lumen of the xenograft of 2 representative uninfected SCID-HU-INT animals (●) and 2 representative animals with human xenografts infected with E. histolytica (□). Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 4 Epithelial barrier integrity is maintained in E. histolytica–infected human intestinal xenografts treated with antisense oligonucleotide to the p65 subunit of human NF-κB. The integrity of the epithelial barrier, as measured by the flux of fluorescently labeled dextrans from the lumen of the human intestinal xenograft into the systemic circulation, is shown for SCID-HU-INT mice whose intestinal xenografts were treated with an intraluminal dose of either nonsense (n = 19) or antisense (n = 13) oligonucleotides before E. histolytica infection (left panel). The flux of dextran was significantly lower (P < 0.05) in E. histolytica–infected human intestinal xenografts that had been pretreated with the antisense oligonucleotide to the p65 subunit of human NF-κB. Treatment of uninfected intestinal xenografts, with either the antisense oligonucleotide (n = 10) or nonsense oligonucleotide (n = 10), had no effect on intestinal xenograft permeability in SCID-HU-INT mice (right panel). Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 5 Epithelial barrier integrity is maintained in E. histolytica–infected human intestinal xenografts in SCID-HU-INT mice treated with the neutrophil-depleting monoclonal antibody RB6-8C5. The barrier function of E. histolytica–infected human intestinal xenografts, as measured by the flow of fluorescently labeled dextran from the lumen of the xenograft to the systemic circulation of SCID-HU-INT mice, was compared in neutrophil-depleted (RB6-8C5-treated) (n = 16) and non–neutrophil-depleted (148-d4-1 treated, n = 12; or untreated, n = 23) SCID-HU-INT mice. The flux of fluorescently labeled dextran from the E. histolytica–infected human intestinal xenograft lumen to the systemic circulation at the 4-hour time point was significantly greater in non–neutrophil-depleted SCID-HU-INT mice than in neutrophil-depleted SCID-HU-INT mice (P < 0.05). Neutrophil depletion by RB6-8C5 (n = 10) or 148-d4-1 treatment (n = 10) had no effect on intestinal permeability in uninfected human intestinal xenografts. Gastroenterology 1998 115, 1446-1453DOI: (10.1016/S0016-5085(98)70023-X) Copyright © 1998 American Gastroenterological Association Terms and Conditions