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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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OVA-expressing E. coli induce clonal expansion of DO11.10 T cells in vivo. DO11.10/BALB/c adoptive transfer recipients were challenged with a single intravenous injection of 106 viable pnir15.OVA-E. coli or pnir15.TET-E. coli. Two-color flow cytometric analysis was performed on splenic or pooled peripheral lymph node lymphocytes isolated at the indicated times after immunization as described in Materials and Methods. The percentages of KJ1–26+CD4+ cells in the lymphocyte gate were determined and used to calculate the total number of splenic KJ1–26+ CD4+ as described previously (reference 36). (A) Representative flow cytometric profiles for lymph node lymphocytes isolated 5 d after challenge with pnir15.TET-E. coli (left) or pnir15.OVA-E. coli (right). The percentages of total gated cells in each quadrant are indicated. (B) Kinetics of clonal expansion of total splenic KJ1–26+CD4+ T cells induced by pnir15.OVA-E. coli versus control pnir15.TET-E. coli. Data points are the mean ± SD of 3–4 animals per group.
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fig2: OVA-expressing E. coli induce clonal expansion of DO11.10 T cells in vivo. DO11.10/BALB/c adoptive transfer recipients were challenged with a single intravenous injection of 106 viable pnir15.OVA-E. coli or pnir15.TET-E. coli. Two-color flow cytometric analysis was performed on splenic or pooled peripheral lymph node lymphocytes isolated at the indicated times after immunization as described in Materials and Methods. The percentages of KJ1–26+CD4+ cells in the lymphocyte gate were determined and used to calculate the total number of splenic KJ1–26+ CD4+ as described previously (reference 36). (A) Representative flow cytometric profiles for lymph node lymphocytes isolated 5 d after challenge with pnir15.TET-E. coli (left) or pnir15.OVA-E. coli (right). The percentages of total gated cells in each quadrant are indicated. (B) Kinetics of clonal expansion of total splenic KJ1–26+CD4+ T cells induced by pnir15.OVA-E. coli versus control pnir15.TET-E. coli. Data points are the mean ± SD of 3–4 animals per group.

Mentions: The immunogenicity of live pnir15.OVA-E. coli was assessed in vivo using a DO11.10 adoptive transfer system (45). Normal BALB/c mice that received transfers of 5 × 106 DO11.10 CD4+ T cells (DO11.10/BALB mice) were innoculated intravenously with 106 pnir15.OVA-E. coli or pnir15.TET-E. coli, and expansion of the clonotypic T cell population in the spleens of recipient mice was monitored by flow cytometric analysis (Fig. 2) . In agreement with a previous study by Chen et al. (46), intravenous administration of the OVA-producing E. coli stimulated a robust clonal expansion in the spleens and lymph nodes of DO11.10 adoptive transfer recipients. Significant clonal expansion (approximately fivefold) was induced by challenge with pnir15.OVA-E. coli, but not the pnir15.TET-E. coli control. The peak response measured was on day five, with a rapid decline to baseline by day nine. Induction of a comparable peak response with free OVA required intraperitoneal administration of a ∼100-fold higher dose of OVA (100 μg) in complete Freund's or a block copolymer adjuvant (data not shown), indicating that the bacterially associated OVA is more efficient at stimulating an in vivo response. Collectively, these data indicate that the pnir15.OVA plasmid produces high-level expression of OVA that is potently immunogenic for DO11.10 T cells in vitro and in vivo.


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)

OVA-expressing E. coli induce clonal expansion of DO11.10 T cells in vivo. DO11.10/BALB/c adoptive transfer recipients were challenged with a single intravenous injection of 106 viable pnir15.OVA-E. coli or pnir15.TET-E. coli. Two-color flow cytometric analysis was performed on splenic or pooled peripheral lymph node lymphocytes isolated at the indicated times after immunization as described in Materials and Methods. The percentages of KJ1–26+CD4+ cells in the lymphocyte gate were determined and used to calculate the total number of splenic KJ1–26+ CD4+ as described previously (reference 36). (A) Representative flow cytometric profiles for lymph node lymphocytes isolated 5 d after challenge with pnir15.TET-E. coli (left) or pnir15.OVA-E. coli (right). The percentages of total gated cells in each quadrant are indicated. (B) Kinetics of clonal expansion of total splenic KJ1–26+CD4+ T cells induced by pnir15.OVA-E. coli versus control pnir15.TET-E. coli. Data points are the mean ± SD of 3–4 animals per group.
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Related In: Results  -  Collection

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fig2: OVA-expressing E. coli induce clonal expansion of DO11.10 T cells in vivo. DO11.10/BALB/c adoptive transfer recipients were challenged with a single intravenous injection of 106 viable pnir15.OVA-E. coli or pnir15.TET-E. coli. Two-color flow cytometric analysis was performed on splenic or pooled peripheral lymph node lymphocytes isolated at the indicated times after immunization as described in Materials and Methods. The percentages of KJ1–26+CD4+ cells in the lymphocyte gate were determined and used to calculate the total number of splenic KJ1–26+ CD4+ as described previously (reference 36). (A) Representative flow cytometric profiles for lymph node lymphocytes isolated 5 d after challenge with pnir15.TET-E. coli (left) or pnir15.OVA-E. coli (right). The percentages of total gated cells in each quadrant are indicated. (B) Kinetics of clonal expansion of total splenic KJ1–26+CD4+ T cells induced by pnir15.OVA-E. coli versus control pnir15.TET-E. coli. Data points are the mean ± SD of 3–4 animals per group.
Mentions: The immunogenicity of live pnir15.OVA-E. coli was assessed in vivo using a DO11.10 adoptive transfer system (45). Normal BALB/c mice that received transfers of 5 × 106 DO11.10 CD4+ T cells (DO11.10/BALB mice) were innoculated intravenously with 106 pnir15.OVA-E. coli or pnir15.TET-E. coli, and expansion of the clonotypic T cell population in the spleens of recipient mice was monitored by flow cytometric analysis (Fig. 2) . In agreement with a previous study by Chen et al. (46), intravenous administration of the OVA-producing E. coli stimulated a robust clonal expansion in the spleens and lymph nodes of DO11.10 adoptive transfer recipients. Significant clonal expansion (approximately fivefold) was induced by challenge with pnir15.OVA-E. coli, but not the pnir15.TET-E. coli control. The peak response measured was on day five, with a rapid decline to baseline by day nine. Induction of a comparable peak response with free OVA required intraperitoneal administration of a ∼100-fold higher dose of OVA (100 μg) in complete Freund's or a block copolymer adjuvant (data not shown), indicating that the bacterially associated OVA is more efficient at stimulating an in vivo response. Collectively, these data indicate that the pnir15.OVA plasmid produces high-level expression of OVA that is potently immunogenic for DO11.10 T cells in vitro and in vivo.

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