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T helper 1 and T helper 2 cells are pathogenic in an antigen-specific model of colitis.

Iqbal N, Oliver JR, Wagner FH, Lazenby AS, Elson CO, Weaver CT - J. Exp. Med. (2002)

Bottom Line: Transfer of antigen-naive DO11.RAG-2(-/-) T cells into recipients colonized with OVA-E. coli resulted in enhanced intestinal recruitment and cell cycling of OVA-specific T cells; however, there was no development of disease.The histopathologic features of disease induced by Th1 and Th2 transfers were distinct, but disease severity was comparable.Induction of disease by both Th1 and Th2 transfers was dependent on bacterially associated OVA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL 35233, USA.

ABSTRACT
Dysregulated T cell responses to enteric bacteria have been implicated as a common mechanism underlying pathogenesis in rodent models of colitis. However, the bacterial species and T cell specificities that induce disease have been poorly defined. We have developed a model system in which target antigen, bacterial host, and corresponding T cell specificity are defined. OVA-specific T cells from DO11.RAG-2(-/-) TCR transgenic mice were transferred into RAG-2(-/-) recipients whose intestinal tracts were colonized with OVA-expressing or control Escherichia coli. Transfer of antigen-naive DO11.RAG-2(-/-) T cells into recipients colonized with OVA-E. coli resulted in enhanced intestinal recruitment and cell cycling of OVA-specific T cells; however, there was no development of disease. In contrast, transfer of polarized T helper (Th) 1 and Th2 populations resulted in severe wasting and colitis in recipients colonized with OVA-expressing but not control E. coli. The histopathologic features of disease induced by Th1 and Th2 transfers were distinct, but disease severity was comparable. Induction of disease by both Th1 and Th2 transfers was dependent on bacterially associated OVA. These results establish that a single bacterially associated antigen can drive the progression of colitis mediated by both Th1 and Th2 cells and provide a new model for understanding the immunoregulatory interactions between T cells responsive to gut floral antigens.

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Th1 and Th2 cells derived from DO11.RAG-2−/− mice induce chronic wasting in RAG-2−/− recipients colonized with pnir15.OVA-E. coli. (A) Cytokine analysis of naive and Th1- or Th2-polarized OVA-specific T cells. Th1 and Th2 cells were generated in vitro under polarizing cytokine conditions as described in Materials and Methods. Briefly, CD4+ T cells isolated from DO11.RAG-2−/− were cultured for 7 d with OVA323–339 peptide, irradiated BALB/c splenocytes, and either IL-12 plus anti–IL-4 (Th1) or IL-4 plus anti-IL-12 (Th2). To confirm Th1/Th2 polarization, aliquots were restimulated for 6 h with PMA and ionomycin, and analyzed for cytokine mRNA expression by ribonuclease protection assay (RPA; mCK-1 RiboQuant kit; BD PharMingen) in comparison with naive T cell isolates. (B) Weight changes of BALB.RAG-2−/− mice that were colonized with 1010 CFU pnir15.OVA-E. coli (black symbols) or pnir15.TET-E. coli (white symbols), reconstituted with 106 naive (○), Th1 (▿), or Th2 (□) cells from DO11.RAG-2−/− donors and monitored for weight loss (B). Data are the mean weekly weights and SD (n = 5–6 mice per group).
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fig3: Th1 and Th2 cells derived from DO11.RAG-2−/− mice induce chronic wasting in RAG-2−/− recipients colonized with pnir15.OVA-E. coli. (A) Cytokine analysis of naive and Th1- or Th2-polarized OVA-specific T cells. Th1 and Th2 cells were generated in vitro under polarizing cytokine conditions as described in Materials and Methods. Briefly, CD4+ T cells isolated from DO11.RAG-2−/− were cultured for 7 d with OVA323–339 peptide, irradiated BALB/c splenocytes, and either IL-12 plus anti–IL-4 (Th1) or IL-4 plus anti-IL-12 (Th2). To confirm Th1/Th2 polarization, aliquots were restimulated for 6 h with PMA and ionomycin, and analyzed for cytokine mRNA expression by ribonuclease protection assay (RPA; mCK-1 RiboQuant kit; BD PharMingen) in comparison with naive T cell isolates. (B) Weight changes of BALB.RAG-2−/− mice that were colonized with 1010 CFU pnir15.OVA-E. coli (black symbols) or pnir15.TET-E. coli (white symbols), reconstituted with 106 naive (○), Th1 (▿), or Th2 (□) cells from DO11.RAG-2−/− donors and monitored for weight loss (B). Data are the mean weekly weights and SD (n = 5–6 mice per group).

Mentions: To evaluate the in vivo responses of antigen-specific T cells to a floral, bacterially associated antigen, OVA-specific T cells were transferred into congenic, immunodeficient recipients whose intestinal tracts were colonized with E. coli that expressed either OVA (pnir15.OVA-E. coli) or a control antigen, TET (pnir15.TET-E. coli). Three T cell populations were examined: naive CD4+ T cells or Th1 and Th2 cells derived by short-term in vitro culture under selective cytokine conditions. A 1 wk protocol was used for Th1/Th2 derivation because it induced excellent polarization of cytokine phenotype (Fig. 3 A), but did not significantly affect T cell migration to the secondary lymphoid organs compared with Th1 and Th2 cells maintained in vitro for longer periods of time (unpublished data). In previous studies, we found that a fraction of CD4+ T cells that expressed the transgenic TCR in DO11.10 mice coexpressed a second, endogenous TCR that recognized bacterial antigens in the normal flora (35). Hence, the DO11.10 donors used for these experiments were crossed onto a BALB.RAG-2−/− background (DO11.RAG-2−/−) to obviate possible complications due to dual TCR specificities. In preliminary colonization studies, it was found that delivery of 1010 anaerobically induced organisms by gastric gavage resulted in a transient colonization of the colon (half-life of 3–4 d; data not shown). Biweekly gavage maintained the floral colony count at a minimum of 108 CFU per gram of colonic contents and was used in experimental protocols.


T helper 1 and T helper 2 cells are pathogenic in an antigen-specific model of colitis.

Iqbal N, Oliver JR, Wagner FH, Lazenby AS, Elson CO, Weaver CT - J. Exp. Med. (2002)

Th1 and Th2 cells derived from DO11.RAG-2−/− mice induce chronic wasting in RAG-2−/− recipients colonized with pnir15.OVA-E. coli. (A) Cytokine analysis of naive and Th1- or Th2-polarized OVA-specific T cells. Th1 and Th2 cells were generated in vitro under polarizing cytokine conditions as described in Materials and Methods. Briefly, CD4+ T cells isolated from DO11.RAG-2−/− were cultured for 7 d with OVA323–339 peptide, irradiated BALB/c splenocytes, and either IL-12 plus anti–IL-4 (Th1) or IL-4 plus anti-IL-12 (Th2). To confirm Th1/Th2 polarization, aliquots were restimulated for 6 h with PMA and ionomycin, and analyzed for cytokine mRNA expression by ribonuclease protection assay (RPA; mCK-1 RiboQuant kit; BD PharMingen) in comparison with naive T cell isolates. (B) Weight changes of BALB.RAG-2−/− mice that were colonized with 1010 CFU pnir15.OVA-E. coli (black symbols) or pnir15.TET-E. coli (white symbols), reconstituted with 106 naive (○), Th1 (▿), or Th2 (□) cells from DO11.RAG-2−/− donors and monitored for weight loss (B). Data are the mean weekly weights and SD (n = 5–6 mice per group).
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Related In: Results  -  Collection

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fig3: Th1 and Th2 cells derived from DO11.RAG-2−/− mice induce chronic wasting in RAG-2−/− recipients colonized with pnir15.OVA-E. coli. (A) Cytokine analysis of naive and Th1- or Th2-polarized OVA-specific T cells. Th1 and Th2 cells were generated in vitro under polarizing cytokine conditions as described in Materials and Methods. Briefly, CD4+ T cells isolated from DO11.RAG-2−/− were cultured for 7 d with OVA323–339 peptide, irradiated BALB/c splenocytes, and either IL-12 plus anti–IL-4 (Th1) or IL-4 plus anti-IL-12 (Th2). To confirm Th1/Th2 polarization, aliquots were restimulated for 6 h with PMA and ionomycin, and analyzed for cytokine mRNA expression by ribonuclease protection assay (RPA; mCK-1 RiboQuant kit; BD PharMingen) in comparison with naive T cell isolates. (B) Weight changes of BALB.RAG-2−/− mice that were colonized with 1010 CFU pnir15.OVA-E. coli (black symbols) or pnir15.TET-E. coli (white symbols), reconstituted with 106 naive (○), Th1 (▿), or Th2 (□) cells from DO11.RAG-2−/− donors and monitored for weight loss (B). Data are the mean weekly weights and SD (n = 5–6 mice per group).
Mentions: To evaluate the in vivo responses of antigen-specific T cells to a floral, bacterially associated antigen, OVA-specific T cells were transferred into congenic, immunodeficient recipients whose intestinal tracts were colonized with E. coli that expressed either OVA (pnir15.OVA-E. coli) or a control antigen, TET (pnir15.TET-E. coli). Three T cell populations were examined: naive CD4+ T cells or Th1 and Th2 cells derived by short-term in vitro culture under selective cytokine conditions. A 1 wk protocol was used for Th1/Th2 derivation because it induced excellent polarization of cytokine phenotype (Fig. 3 A), but did not significantly affect T cell migration to the secondary lymphoid organs compared with Th1 and Th2 cells maintained in vitro for longer periods of time (unpublished data). In previous studies, we found that a fraction of CD4+ T cells that expressed the transgenic TCR in DO11.10 mice coexpressed a second, endogenous TCR that recognized bacterial antigens in the normal flora (35). Hence, the DO11.10 donors used for these experiments were crossed onto a BALB.RAG-2−/− background (DO11.RAG-2−/−) to obviate possible complications due to dual TCR specificities. In preliminary colonization studies, it was found that delivery of 1010 anaerobically induced organisms by gastric gavage resulted in a transient colonization of the colon (half-life of 3–4 d; data not shown). Biweekly gavage maintained the floral colony count at a minimum of 108 CFU per gram of colonic contents and was used in experimental protocols.

Bottom Line: Transfer of antigen-naive DO11.RAG-2(-/-) T cells into recipients colonized with OVA-E. coli resulted in enhanced intestinal recruitment and cell cycling of OVA-specific T cells; however, there was no development of disease.The histopathologic features of disease induced by Th1 and Th2 transfers were distinct, but disease severity was comparable.Induction of disease by both Th1 and Th2 transfers was dependent on bacterially associated OVA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL 35233, USA.

ABSTRACT
Dysregulated T cell responses to enteric bacteria have been implicated as a common mechanism underlying pathogenesis in rodent models of colitis. However, the bacterial species and T cell specificities that induce disease have been poorly defined. We have developed a model system in which target antigen, bacterial host, and corresponding T cell specificity are defined. OVA-specific T cells from DO11.RAG-2(-/-) TCR transgenic mice were transferred into RAG-2(-/-) recipients whose intestinal tracts were colonized with OVA-expressing or control Escherichia coli. Transfer of antigen-naive DO11.RAG-2(-/-) T cells into recipients colonized with OVA-E. coli resulted in enhanced intestinal recruitment and cell cycling of OVA-specific T cells; however, there was no development of disease. In contrast, transfer of polarized T helper (Th) 1 and Th2 populations resulted in severe wasting and colitis in recipients colonized with OVA-expressing but not control E. coli. The histopathologic features of disease induced by Th1 and Th2 transfers were distinct, but disease severity was comparable. Induction of disease by both Th1 and Th2 transfers was dependent on bacterially associated OVA. These results establish that a single bacterially associated antigen can drive the progression of colitis mediated by both Th1 and Th2 cells and provide a new model for understanding the immunoregulatory interactions between T cells responsive to gut floral antigens.

Show MeSH
Related in: MedlinePlus