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A critical role for CD40-CD40 ligand interactions in amplification of the mucosal CD8 T cell response.

Lefrançois L, Olson S, Masopust D - J. Exp. Med. (1999)

Bottom Line: Appearance of virus-specific mucosal, but not splenic, CD8 cells also relied heavily on CD40-CD40L interactions.The mucosal CTL response of transferred CD8 T cells was MHC class II and interleukin 12 independent.The results established a novel pathway of direct CD40L-mediated CD8 T cell activation.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatic Diseases, University of Connecticut Health Center, Farmington, Connecticut 06030, USA. llefranc@panda.uchc.edu

ABSTRACT
The role of CD40 ligand (CD40L) in CD8 T cell activation was assessed by tracking antigen-specific T cells in vivo using both adoptive transfer of T cell receptor transgenic T cells and major histocompatibility complex (MHC) class I tetramers. Soluble antigen immunization induced entry of CD8 cells into the intestinal mucosa and cytotoxic T lymphocyte (CTL) differentiation, whereas CD8 cells in secondary lymphoid tissue proliferated but were not cytolytic. Immunization concurrent with CD40L blockade or in the absence of CD40 demonstrated that accumulation of CD8 T cells in the mucosa was CD40L dependent. Furthermore, activation was mediated through CD40L expressed by the CD8 cells, since inhibition by anti-CD40L monoclonal antibodies occurred after adoptive transfer to CD40L-deficient mice. However, mucosal CD8 T cells in normal and CD40(-/-) mice were equivalent killers, indicating that CD40L was not required for CTL differentiation. Appearance of virus-specific mucosal, but not splenic, CD8 cells also relied heavily on CD40-CD40L interactions. The mucosal CTL response of transferred CD8 T cells was MHC class II and interleukin 12 independent. The results established a novel pathway of direct CD40L-mediated CD8 T cell activation.

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Direct inhibition of T cell expansion by CD8 T cell CD40L blockade. 2.5 × 106 naive OT-I–RAG−/− cells (Ly5.2) were transferred to Ly5.1 CD40L−/− mice. Mice were treated daily with αCD40L or control antibody starting at 1 d before immunization. 2 d after transfer mice were immunized with 5 mg OVA, and 3 d later MLN and LP cells were isolated and analyzed by flow cytometry for the presence of donor cells by analysis of Ly5.2 and CD8 expression. In unimmunized mice, OT-I cells comprised 0.4% of MLN cells and were undetectable in LP cells (data not shown).
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Figure 3: Direct inhibition of T cell expansion by CD8 T cell CD40L blockade. 2.5 × 106 naive OT-I–RAG−/− cells (Ly5.2) were transferred to Ly5.1 CD40L−/− mice. Mice were treated daily with αCD40L or control antibody starting at 1 d before immunization. 2 d after transfer mice were immunized with 5 mg OVA, and 3 d later MLN and LP cells were isolated and analyzed by flow cytometry for the presence of donor cells by analysis of Ly5.2 and CD8 expression. In unimmunized mice, OT-I cells comprised 0.4% of MLN cells and were undetectable in LP cells (data not shown).

Mentions: The results thus far demonstrated a role for CD40L in CD8 T cell activation, but did not indicate whether this effect was direct or indirect. To test this, we transferred OT-I–RAG−/− cells into CD40L-deficient mice and attempted to block activation with MR1. In this experiment, only the transferred CD8 T cells were capable of expressing CD40L, so that any inhibitory effect must be mediated at the level of the OT-I cells. After OT-I cell transfer, immunization of CD40L−/− mice with sOVA resulted in substantial proliferation in the periphery and appearance of OT-I cells in the LP (Fig. 3). Treatment with anti-CD40L mAb resulted in partial inhibition of OT-I accumulation in MLNs and much greater inhibition of OT-I accumulation in the LP (Fig. 3). Little inhibition of OT-I expansion in PLNs was observed (data not shown). These results were similar to those obtained in CD40L-competent mice (Fig. 1 and Fig. 2), and indicated that CD40L expressed by CD8 T cells was responsible for activation of OT-I cells destined for the mucosa.


A critical role for CD40-CD40 ligand interactions in amplification of the mucosal CD8 T cell response.

Lefrançois L, Olson S, Masopust D - J. Exp. Med. (1999)

Direct inhibition of T cell expansion by CD8 T cell CD40L blockade. 2.5 × 106 naive OT-I–RAG−/− cells (Ly5.2) were transferred to Ly5.1 CD40L−/− mice. Mice were treated daily with αCD40L or control antibody starting at 1 d before immunization. 2 d after transfer mice were immunized with 5 mg OVA, and 3 d later MLN and LP cells were isolated and analyzed by flow cytometry for the presence of donor cells by analysis of Ly5.2 and CD8 expression. In unimmunized mice, OT-I cells comprised 0.4% of MLN cells and were undetectable in LP cells (data not shown).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195681&req=5

Figure 3: Direct inhibition of T cell expansion by CD8 T cell CD40L blockade. 2.5 × 106 naive OT-I–RAG−/− cells (Ly5.2) were transferred to Ly5.1 CD40L−/− mice. Mice were treated daily with αCD40L or control antibody starting at 1 d before immunization. 2 d after transfer mice were immunized with 5 mg OVA, and 3 d later MLN and LP cells were isolated and analyzed by flow cytometry for the presence of donor cells by analysis of Ly5.2 and CD8 expression. In unimmunized mice, OT-I cells comprised 0.4% of MLN cells and were undetectable in LP cells (data not shown).
Mentions: The results thus far demonstrated a role for CD40L in CD8 T cell activation, but did not indicate whether this effect was direct or indirect. To test this, we transferred OT-I–RAG−/− cells into CD40L-deficient mice and attempted to block activation with MR1. In this experiment, only the transferred CD8 T cells were capable of expressing CD40L, so that any inhibitory effect must be mediated at the level of the OT-I cells. After OT-I cell transfer, immunization of CD40L−/− mice with sOVA resulted in substantial proliferation in the periphery and appearance of OT-I cells in the LP (Fig. 3). Treatment with anti-CD40L mAb resulted in partial inhibition of OT-I accumulation in MLNs and much greater inhibition of OT-I accumulation in the LP (Fig. 3). Little inhibition of OT-I expansion in PLNs was observed (data not shown). These results were similar to those obtained in CD40L-competent mice (Fig. 1 and Fig. 2), and indicated that CD40L expressed by CD8 T cells was responsible for activation of OT-I cells destined for the mucosa.

Bottom Line: Appearance of virus-specific mucosal, but not splenic, CD8 cells also relied heavily on CD40-CD40L interactions.The mucosal CTL response of transferred CD8 T cells was MHC class II and interleukin 12 independent.The results established a novel pathway of direct CD40L-mediated CD8 T cell activation.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatic Diseases, University of Connecticut Health Center, Farmington, Connecticut 06030, USA. llefranc@panda.uchc.edu

ABSTRACT
The role of CD40 ligand (CD40L) in CD8 T cell activation was assessed by tracking antigen-specific T cells in vivo using both adoptive transfer of T cell receptor transgenic T cells and major histocompatibility complex (MHC) class I tetramers. Soluble antigen immunization induced entry of CD8 cells into the intestinal mucosa and cytotoxic T lymphocyte (CTL) differentiation, whereas CD8 cells in secondary lymphoid tissue proliferated but were not cytolytic. Immunization concurrent with CD40L blockade or in the absence of CD40 demonstrated that accumulation of CD8 T cells in the mucosa was CD40L dependent. Furthermore, activation was mediated through CD40L expressed by the CD8 cells, since inhibition by anti-CD40L monoclonal antibodies occurred after adoptive transfer to CD40L-deficient mice. However, mucosal CD8 T cells in normal and CD40(-/-) mice were equivalent killers, indicating that CD40L was not required for CTL differentiation. Appearance of virus-specific mucosal, but not splenic, CD8 cells also relied heavily on CD40-CD40L interactions. The mucosal CTL response of transferred CD8 T cells was MHC class II and interleukin 12 independent. The results established a novel pathway of direct CD40L-mediated CD8 T cell activation.

Show MeSH
Related in: MedlinePlus