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Bacteria-triggered CD4(+) T regulatory cells suppress Helicobacter hepaticus-induced colitis.

Kullberg MC, Jankovic D, Gorelick PL, Caspar P, Letterio JJ, Cheever AW, Sher A - J. Exp. Med. (2002)

Bottom Line: The disease-protective WT CD4(+) cells are contained within the CD45RB(low) fraction and unexpectedly were found in both the CD25(+) and the CD25(-) subpopulations of these cells, their frequency being higher in the latter.The mechanism by which CD25(+) and CD25(-) CD45RB(low) CD4(+) cells block colitis involves IL-10 and not transforming growth factor (TGF)-beta, as treatment with anti-IL-10R but not anti-TGF-beta monoclonal antibody abrogated their protective effect.The induction of such cells in response to gut flora may be a mechanism protecting normal individuals against inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Immunobiology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases/NIH, Building 50, Room 6146, 50 South Drive, Bethesda, MD 20892, USA. MKullberg@niaid.nih.gov

ABSTRACT
We have previously demonstrated that interleukin (IL)-10-deficient (IL-10 knockout [KO]) but not wild-type (WT) mice develop colitis after infection with Helicobacter hepaticus. Here, we show that infected recombination activating gene (RAG) KO mice develop intestinal inflammation after reconstitution with CD4(+) T cells from IL-10 KO animals and that the cotransfer of CD4(+) T cells from H. hepaticus-infected but not uninfected WT mice prevents this colitis. The disease-protective WT CD4(+) cells are contained within the CD45RB(low) fraction and unexpectedly were found in both the CD25(+) and the CD25(-) subpopulations of these cells, their frequency being higher in the latter. The mechanism by which CD25(+) and CD25(-) CD45RB(low) CD4(+) cells block colitis involves IL-10 and not transforming growth factor (TGF)-beta, as treatment with anti-IL-10R but not anti-TGF-beta monoclonal antibody abrogated their protective effect. In vitro, CD45RB(low) CD4(+) cells from infected WT mice were shown to produce IL-10 and suppress interferon-gamma production by IL-10 KO CD4(+) cells in an H. hepaticus antigen-specific manner. Together, our data support the concept that H. hepaticus infection results in the induction in WT mice of regulatory T cells that prevent bacteria-induced colitis. The induction of such cells in response to gut flora may be a mechanism protecting normal individuals against inflammatory bowel disease.

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H. hepaticus–infected but not naive RAG KO mice develop intestinal inflammation after reconstitution with CD4+ T cells from IL-10 KO mice. (A) Uninfected or H. hepaticus–infected RAG KO mice were inoculated intravenously with 3 × 105 CD4+ cells from the MLNs of 11-wk infected IL-10 KO mice, and intestinal pathology was analyzed 8, 15, 22, and 29 d later (solid bars). Naive and infected RAG KO animals receiving no cells were analyzed in parallel (open bars). Bars represent mean cecal histology scores ± SD of three or four mice per group. *, P < 0.05 compared with infected mice receiving no cells. (B–D) Histology of ceca of representative sections from the mice shown in A analyzed 29 d after cell transfer: (B) infected RAG KO without cell transfer; (C) infected RAG KO receiving CD4+ cells from infected IL-10 KO mice; or (D) uninfected RAG KO receiving CD4+ cells from infected IL-10 KO mice. Crypt abscesses (solid arrows), ulcer (open arrow). H&E staining. Bar, 200 μm.
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fig1: H. hepaticus–infected but not naive RAG KO mice develop intestinal inflammation after reconstitution with CD4+ T cells from IL-10 KO mice. (A) Uninfected or H. hepaticus–infected RAG KO mice were inoculated intravenously with 3 × 105 CD4+ cells from the MLNs of 11-wk infected IL-10 KO mice, and intestinal pathology was analyzed 8, 15, 22, and 29 d later (solid bars). Naive and infected RAG KO animals receiving no cells were analyzed in parallel (open bars). Bars represent mean cecal histology scores ± SD of three or four mice per group. *, P < 0.05 compared with infected mice receiving no cells. (B–D) Histology of ceca of representative sections from the mice shown in A analyzed 29 d after cell transfer: (B) infected RAG KO without cell transfer; (C) infected RAG KO receiving CD4+ cells from infected IL-10 KO mice; or (D) uninfected RAG KO receiving CD4+ cells from infected IL-10 KO mice. Crypt abscesses (solid arrows), ulcer (open arrow). H&E staining. Bar, 200 μm.

Mentions: We have previously shown that SPF-reared IL-10 KO mice develop a Th1 cytokine-associated colitis after experimental infection with H. hepaticus (20, 21). In contrast, H. hepaticus–inoculated WT mice are free of disease and mount a T cell–dependent IL-10 response to the bacterium (20), suggesting the development of a disease-protective CD4+ T cell response. To characterize this protective T cell response in WT mice, we used a modification of the CD4/SCID transfer model of colitis involving experimental infection with bacteria in which disease is induced in H. hepaticus–infected RAG KO recipients by the transfer of CD4+ cells from infected colitic IL-10 KO mice. After inoculation with H. hepaticus, RAG KO mice on a C57BL/10 background showed no or minimal signs of intestinal inflammation when examined up to 34 wk after infection (Fig. 1, A and B , and not depicted). However, if 2–4-d infected RAG KO mice were reconstituted with MLN CD4+ cells from infected IL-10 KO animals, inflammation developed in the large intestine (colitis) including the cecum (typhlitis) within 1 wk, gradually increasing over a 4-wk period (Fig. 1 A). Similar results were observed when CD4+ cells from naive IL-10 KO mice were transferred to infected RAG KO recipients (unpublished data), a result we interpret as reflecting the sensitization of the naive transferred CD4+ cells by H. hepaticus in the infected recipients. The colitis observed in the RAG KO recipients was characterized by mucosal hyperplasia and inflammatory cell infiltrates in the lamina propria, submucosa, and serosa, as well as by the presence, in severe cases, of crypt abscesses and ulcers (Fig. 1 C). Importantly, when uninfected RAG KO mice were given IL-10 KO CD4+ cells, no inflammation was observed in the same time period (Fig. 1, A and D). These results demonstrate a critical role for both T cells and the Helicobacter organism in disease induction.


Bacteria-triggered CD4(+) T regulatory cells suppress Helicobacter hepaticus-induced colitis.

Kullberg MC, Jankovic D, Gorelick PL, Caspar P, Letterio JJ, Cheever AW, Sher A - J. Exp. Med. (2002)

H. hepaticus–infected but not naive RAG KO mice develop intestinal inflammation after reconstitution with CD4+ T cells from IL-10 KO mice. (A) Uninfected or H. hepaticus–infected RAG KO mice were inoculated intravenously with 3 × 105 CD4+ cells from the MLNs of 11-wk infected IL-10 KO mice, and intestinal pathology was analyzed 8, 15, 22, and 29 d later (solid bars). Naive and infected RAG KO animals receiving no cells were analyzed in parallel (open bars). Bars represent mean cecal histology scores ± SD of three or four mice per group. *, P < 0.05 compared with infected mice receiving no cells. (B–D) Histology of ceca of representative sections from the mice shown in A analyzed 29 d after cell transfer: (B) infected RAG KO without cell transfer; (C) infected RAG KO receiving CD4+ cells from infected IL-10 KO mice; or (D) uninfected RAG KO receiving CD4+ cells from infected IL-10 KO mice. Crypt abscesses (solid arrows), ulcer (open arrow). H&E staining. Bar, 200 μm.
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fig1: H. hepaticus–infected but not naive RAG KO mice develop intestinal inflammation after reconstitution with CD4+ T cells from IL-10 KO mice. (A) Uninfected or H. hepaticus–infected RAG KO mice were inoculated intravenously with 3 × 105 CD4+ cells from the MLNs of 11-wk infected IL-10 KO mice, and intestinal pathology was analyzed 8, 15, 22, and 29 d later (solid bars). Naive and infected RAG KO animals receiving no cells were analyzed in parallel (open bars). Bars represent mean cecal histology scores ± SD of three or four mice per group. *, P < 0.05 compared with infected mice receiving no cells. (B–D) Histology of ceca of representative sections from the mice shown in A analyzed 29 d after cell transfer: (B) infected RAG KO without cell transfer; (C) infected RAG KO receiving CD4+ cells from infected IL-10 KO mice; or (D) uninfected RAG KO receiving CD4+ cells from infected IL-10 KO mice. Crypt abscesses (solid arrows), ulcer (open arrow). H&E staining. Bar, 200 μm.
Mentions: We have previously shown that SPF-reared IL-10 KO mice develop a Th1 cytokine-associated colitis after experimental infection with H. hepaticus (20, 21). In contrast, H. hepaticus–inoculated WT mice are free of disease and mount a T cell–dependent IL-10 response to the bacterium (20), suggesting the development of a disease-protective CD4+ T cell response. To characterize this protective T cell response in WT mice, we used a modification of the CD4/SCID transfer model of colitis involving experimental infection with bacteria in which disease is induced in H. hepaticus–infected RAG KO recipients by the transfer of CD4+ cells from infected colitic IL-10 KO mice. After inoculation with H. hepaticus, RAG KO mice on a C57BL/10 background showed no or minimal signs of intestinal inflammation when examined up to 34 wk after infection (Fig. 1, A and B , and not depicted). However, if 2–4-d infected RAG KO mice were reconstituted with MLN CD4+ cells from infected IL-10 KO animals, inflammation developed in the large intestine (colitis) including the cecum (typhlitis) within 1 wk, gradually increasing over a 4-wk period (Fig. 1 A). Similar results were observed when CD4+ cells from naive IL-10 KO mice were transferred to infected RAG KO recipients (unpublished data), a result we interpret as reflecting the sensitization of the naive transferred CD4+ cells by H. hepaticus in the infected recipients. The colitis observed in the RAG KO recipients was characterized by mucosal hyperplasia and inflammatory cell infiltrates in the lamina propria, submucosa, and serosa, as well as by the presence, in severe cases, of crypt abscesses and ulcers (Fig. 1 C). Importantly, when uninfected RAG KO mice were given IL-10 KO CD4+ cells, no inflammation was observed in the same time period (Fig. 1, A and D). These results demonstrate a critical role for both T cells and the Helicobacter organism in disease induction.

Bottom Line: The disease-protective WT CD4(+) cells are contained within the CD45RB(low) fraction and unexpectedly were found in both the CD25(+) and the CD25(-) subpopulations of these cells, their frequency being higher in the latter.The mechanism by which CD25(+) and CD25(-) CD45RB(low) CD4(+) cells block colitis involves IL-10 and not transforming growth factor (TGF)-beta, as treatment with anti-IL-10R but not anti-TGF-beta monoclonal antibody abrogated their protective effect.The induction of such cells in response to gut flora may be a mechanism protecting normal individuals against inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Immunobiology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases/NIH, Building 50, Room 6146, 50 South Drive, Bethesda, MD 20892, USA. MKullberg@niaid.nih.gov

ABSTRACT
We have previously demonstrated that interleukin (IL)-10-deficient (IL-10 knockout [KO]) but not wild-type (WT) mice develop colitis after infection with Helicobacter hepaticus. Here, we show that infected recombination activating gene (RAG) KO mice develop intestinal inflammation after reconstitution with CD4(+) T cells from IL-10 KO animals and that the cotransfer of CD4(+) T cells from H. hepaticus-infected but not uninfected WT mice prevents this colitis. The disease-protective WT CD4(+) cells are contained within the CD45RB(low) fraction and unexpectedly were found in both the CD25(+) and the CD25(-) subpopulations of these cells, their frequency being higher in the latter. The mechanism by which CD25(+) and CD25(-) CD45RB(low) CD4(+) cells block colitis involves IL-10 and not transforming growth factor (TGF)-beta, as treatment with anti-IL-10R but not anti-TGF-beta monoclonal antibody abrogated their protective effect. In vitro, CD45RB(low) CD4(+) cells from infected WT mice were shown to produce IL-10 and suppress interferon-gamma production by IL-10 KO CD4(+) cells in an H. hepaticus antigen-specific manner. Together, our data support the concept that H. hepaticus infection results in the induction in WT mice of regulatory T cells that prevent bacteria-induced colitis. The induction of such cells in response to gut flora may be a mechanism protecting normal individuals against inflammatory bowel disease.

Show MeSH
Related in: MedlinePlus