Volume 122, Issue 5, Pages 1346-1354 (May 2002) Monoassociation of SCID mice with Helicobacter muridarum, but not four other enterics, provokes IBD upon receipt of T cells  Han-Qing Jiang, Natasha Kushnir, M.Christine Thurnheer, Nicolaas A. Bos, John J. Cebra  Gastroenterology  Volume 122, Issue 5, Pages 1346-1354 (May 2002) DOI: 10.1053/gast.2002.32959 Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 1 Histologic evaluation of the colon of conventionally reared and germ-free SCID mice reconstituted with either CD4+ CD45RBhigh T cells from mesenteric lymph nodes of conventionally reared BALB/c mice or no cells after 12 weeks. (A) Conventionally reared SCID mice reconstituted with 5–10 × 105 CD4+ CD45RBhigh T cells; (B) conventionally reared SCID mice with no cells; (C) germ-free SCID mice reconstituted with 5–10 × 105 CD4+ CD45RBhigh T cells; and (D) germ-free SCID with no cells. Massive infiltration of mononuclear cells, elongated, hyperplastic crypts, and dramatically reduced numbers of goblet cells were only observed in conventionally reared SCID mice reconstituted with CD4+ CD45RBhigh T cells. No histopathological changes were found in germ-free SCID mice reconstituted with CD4+ CD45RBhigh T cells, as well as in control conventionally reared and germ-free SCID mice receiving no cells. Tissue sections were stained with H&E. Original magnification is 200×. Gastroenterology 2002 122, 1346-1354DOI: (10.1053/gast.2002.32959) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 2 Formerly germ-free SCID mice monoassociated with H. muridarum developed wasting disease and colitis in 5–6 weeks after reconstitution with CD4+ CD45RBhigh T cells. Germ-free SCID mice were intragastrically inoculated with H. muridarum. Two weeks later, mice (n = 5) were intraperitoneally transferred with 5 × 105 CD4+ CD45RBhigh T cells from mesenteric lymph nodes of conventionally reared BALB/c mice. Recipient mice were weighed on the day before cell transfer and weekly thereafter. As controls, the weight changes of H. muridarum monoassociated SCID mice (n = 6) receiving no cells were monitored. The change in weight is expressed as percentage of the original weight before cell transfer. Data represent the mean values ± SEM. Statistical analysis was performed using Student 2-tailed t test. Significant difference between these 2 groups was observed from 6 weeks on after cell transfer (week 6, P = 0.018; week 7, P = 0.0002). Gastroenterology 2002 122, 1346-1354DOI: (10.1053/gast.2002.32959) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 3 H. muridarum monoassociated SCID mice reconstituted with CD4+ CD45RBhigh T cells developed severe colitis. (A) Histologic analysis of colon shows the dramatically thickened intestinal wall with massive infiltration of mononuclear cells, edema, elongated and hyperplastic crypts, crypt abscesses, and goblet cell depletion. (B) Normal histologic structure of the colon was observed in control H. muridarum monoassociated SCID mice receiving no cells for 10 weeks. The tissue sections were H&E stained. Original magnification is 60×. Gastroenterology 2002 122, 1346-1354DOI: (10.1053/gast.2002.32959) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 4 Histologic evaluation of colons from monoassociated SCID mice with various enteric bacteria. Germ-free SCID mice were monoassociated with SFB, O. anthropi, act A (-) mutant of L. monocytogenes, M. morganii, and H. muridarum, respectively, by intragastrical inoculation. All mice were intraperitoneally transferred with 5–10 × 105 CD4+ CD45RBhigh T cells. Six micrometer sections of the colons were made and examined for inflammatory lesions 7 weeks after cell transfer in H. muridarum mice, 10–12 weeks in each of the other 4 bacteria monoassociated mice. (A) SFB monoassociated mice; (B) act A (-) mutant of L. monocytogenes monoassociated mice; (C) O. anthropi monoassociated mice; and (D) H. muridarum monoassociated mice. The inflammatory lesions were only observed in H. muridarum monoassociated mice, but not in each of the other 4 bacteria monoassociated mice (section from M. morganii mice not shown). The tissue sections were H&E stained. Original magnification is 200×. Gastroenterology 2002 122, 1346-1354DOI: (10.1053/gast.2002.32959) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 5 Expansion of CD4+ T cells in (A) spleen and (B) mesenteric lymph nodes of germ-free SCID, monoassociated SCID, and conventionally reared SCID mice reconstituted with CD4+CD45RBhigh T cells. Seven weeks after cell transfer in H. muridarum mice and 12 weeks in all other mice, splenocytes and mesenteric lymph node cells were isolated and stained with PE conjugated monoclonal antibody against CD4. The total number of CD4+ T cells in each tissue sample was calculated as a ratio of the total yield of the viable cells and the percentage of CD4+ T cells in the population as determined by FACS. In spleen, CD4+ T cells were highly expanded in conventionally reared SCID mice, as well as in H. muridarum monoassociated mice (P > 0.05). Compared with conventionally reared mice, CD4+ T cell outgrowth in all other monoassociated mice was significantly low (P < 0.05). In mesenteric lymph nodes, CD4+ T cell expansion in conventionally reared mice was much higher than that in all monoassociated mice (P < 0.05). Almost no CD4+ T cells were recovered from germ-free SCID mice. Each data bar represents the mean value ± SEM of 3–6 mice in each group. Gastroenterology 2002 122, 1346-1354DOI: (10.1053/gast.2002.32959) Copyright © 2002 American Gastroenterological Association Terms and Conditions