Volume 124, Issue 3, Pages 672-682 (March 2003) T helper type-2 cells induce ileal villus atrophy, goblet cell metaplasia, and wasting disease in T cell–deficient mice  Taeko Dohi, Kohtaro Fujihashi, Toshiya Koga, Yuko Shirai, Yuki I. Kawamura, Chieko Ejima, Rie Kato, Kiyoshi Saitoh, Jerry R. McGhee  Gastroenterology  Volume 124, Issue 3, Pages 672-682 (March 2003) DOI: 10.1053/gast.2003.50092 Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 1 Comparison of wasting disease after adoptive transfer of RBHi T cells. (A) Example of the reanalysis of sorted RBHi T cells used for adoptive transfer. After sorting, PE-labeled CD45RBHi T cells were stained with FITC-labeled anti-TCRγδ Ab, anti-NK1.1 Ab, and Gr-1 Ab. PE-negative cells include photo-bleached cells. The RBHi T cell subset was >99% pure. Similar purity was obtained in RBLo T cells. (B) Weight loss after adoptive transfer of Wt (open circle) and IFN-γ−/− (solid circle) RBHi T cells to TCR−/− recipients. Weight changes of age-matched TCR−/− mice are also shown (diamonds). Data are given as the mean and standard error of the mean of 5 mice. Asterisks show statistically significant differences from recipients of Wt RBHi T cells. (C) IFN-γ−/− RBLo T cell transfer did not induce wasting disease, and treatment with anti–IFN-γ mAb after adoptive transfer did not change wasting disease of TCR−/− recipients of IFN-γ−/− RBHi T cells. SCID recipients of IFN-γ−/− RBHi T cells did not show severe weight loss. Bars indicate mean value. Data are given as the mean and standard error of the mean. Asterisks indicate statistically significant difference. Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 2 H&E-stained sections of the colonic and ileal lesions of TCR−/− and SCID recipient mice. Colitis was severe in the TCR−/− (C) and SCID (G) recipients of Wt RBHi T cells but mild in TCR−/− (E) and SCID (I) recipients of IFN-γ−/− RBHi T cells. No significant change was seen in the ileum from the TCR−/− recipients of Wt RBHi T cells except for mild cell infiltration (D), whereas villus atrophy was evident in the ileum in the TCR−/− recipients of IFN-γ−/− RBHi T cells (F). No evident villus atrophy was seen in the ileum from SCID recipients of Wt (H) or IFN-γ−/− (J) RBHi T cells. Colon and ileum from TCR−/− mice without T cell transfer are shown in (A) and (B), respectively. (Original magnification 10× in pictures of the colon and 20× in those of the ileum.) Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 3 Comparison of parameters for tissue pathology in the ileum of TCR−/− and SCID recipients of RBHi T cells. (A) Height of villi and (B) length of crypts were measured in H&E-stained sections. (C) Length of the proliferating zone was measured after staining with anti-Ki67 Ab as shown in Figure 4. (D) The area occupied by goblet cells was detected by PAS reactivity, like the typical cells shown in Figure 4. The bars represent the mean and standard error of the mean. An asterisk indicates a statistically significant difference from the data without asterisks. Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 4 Histochemical analysis of the ileum of TCR−/− recipients of Wt or IFN-γ−/− RBHi T cells. Paraffin-embedded sections were stained for ALP (in red), for Ki67 antigen (in black), and for mucin by PAS (in purple-red). (Original magnification for ALP and PAS staining, 40×; for Ki67 staining, 10×.) Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 5 Ileal lesions in SCID recipients of B220+ cells and IFN-γ−/− RBHi T cells. SCID recipient mice received an intravenous injection of 2×106 B220+ cells prepared from the mesenteric lymph nodes of Wt mice. Three days later, an aliquot of 1×106 IFN-γ−/− RBHi T cells were transferred to a group of recipients. (A) The height of villi and the PAS-positive areas in the ileum were determined as was done in Figure 3. Blank bars are SCID recipients of only B220+ cells; solid bars are SCID recipients of B220+ cells and IFN-γ−/− RBHi T cells. The bars represent the mean and standard error of the mean of 5 mice. An asterisk indicates a statistically significant difference from the data without asterisks. A separate set of experiments using B cells prepared independently reproduced the same results. (B) Ileal frozen sections were prepared from SCID recipients of B220+ cells (left) or B220+ cells and IFN-γ−/− RBHi T cells (right). The sections were stained with hematoxylin and eosin (top), PAS (middle), or biotin-labeled goat anti-mouse IgA antibody followed by aminocoumarin-labeled streptavidin (bottom). (Original magnification for H&E and PAS staining, 20×; for IgA staining, 10×.) Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions

Fig. 6 Treatment of TCR−/− recipients of IFN-γ−/− RBHi T cells with anti–IL-4 mAb. A dose of 1 mg of anti–IL-4 mAb or control rat IgG was injected weekly after adoptive transfer. Each group included 4 mice. (A) Weight loss was prevented by the treatment. Bars show the mean values. (B) Length of villi and (C) length of proliferating zone were increased in the recipients treated with anti–IL-4 mAb. (D) Goblet-cell area decreased in the recipients treated with anti–IL-4 mAb. Data are shown as the mean and the standard error of the mean. An asterisk indicates a statistically significant difference. Gastroenterology 2003 124, 672-682DOI: (10.1053/gast.2003.50092) Copyright © 2003 American Gastroenterological Association Terms and Conditions