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Mucosal memory CD8⁺ T cells are selected in the periphery by an MHC class I molecule.

Huang Y, Park Y, Wang-Zhu Y, Larange A, Arens R, Bernardo I, Olivares-Villagómez D, Herndler-Brandstetter D, Abraham N, Grubeck-Loebenstein B, Schoenberger SP, Van Kaer L, Kronenberg M, Teitell MA, Cheroutre H - Nat. Immunol. (2011)

Bottom Line: The presence of immune memory at pathogen-entry sites is a prerequisite for protection.Nevertheless, the mechanisms that warrant immunity at peripheral interfaces are not understood.Furthermore, constitutive expression of TL on epithelial cells led to continued selection of mature CD8αβ(+) memory T cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Immunology, La Jolla Institute for Allergy & Immunology, La Jolla, California, USA.

ABSTRACT
The presence of immune memory at pathogen-entry sites is a prerequisite for protection. Nevertheless, the mechanisms that warrant immunity at peripheral interfaces are not understood. Here we show that the nonclassical major histocompatibility complex (MHC) class I molecule thymus leukemia antigen (TL), induced on dendritic cells interacting with CD8αα on activated CD8αβ(+) T cells, mediated affinity-based selection of memory precursor cells. Furthermore, constitutive expression of TL on epithelial cells led to continued selection of mature CD8αβ(+) memory T cells. The memory process driven by TL and CD8αα was essential for the generation of CD8αβ(+) memory T cells in the intestine and the accumulation of highly antigen-sensitive CD8αβ(+) memory T cells that form the first line of defense at the largest entry port for pathogens.

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TL negatively affects memory generation of CD8αβ+ T cells(a) WT or TL- mice were orally infected with 1 × 109 ActA- Lm-OVA. 30 days p.i., splenocytes (SPL) and IEL were isolated for IFN-γ intracellular staining and CD8α cell surface staining after ex vivo re-stimulation with OVA257-264 peptide. Graph depicts pooled data ± s.e.m.. (b) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were adoptively transferred into Ly5.2+ WT or TL- recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. Donor OT-I cells were tracked in the spleen and IEL 2 months p.i. Graph depicts pooled data ± s.e.m.. (c) 1 × 106 naïve Ly5.1+ OT-I cells were transferred into WT or TL-Tg recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. The donor OT-I cells in the spleens and IEL were tracked two months p.i. Graph depicts pooled data ± s.e.m.. (d) 5 × 104 Ly5.1+ naïve OT-I cells were transferred into C57BL/6 mice which were subsequently immunized i.v. with 5 × 105 OVAp-loaded DCs generated from bone marrow cells (BMDC) of WT or TL-Tg mice (top). Alternatively, OT-I cells were primed in vitro with TL negative APC or APC transfected with TL and then transferred to B6 recipients (bottom). Memory OT-I cells in the spleen were analyzed 2 months after DC immunization or after transfer of in vitro activated OT-I cells. Graph depicts pooled data ± s.e.m.. (e) Spleen or mLN DC were sorted based on the CD11c and MHC II expression and assessed for TL expression by flow cytometry. (f) Freshly isolated SPL or mLN DC were activated with CpG for 1 d and then analyzed for TL expression. * P < 0.05 and ** P < 0.001 (unpaired t-test). All data are representative of three independent experiments.
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Figure 1: TL negatively affects memory generation of CD8αβ+ T cells(a) WT or TL- mice were orally infected with 1 × 109 ActA- Lm-OVA. 30 days p.i., splenocytes (SPL) and IEL were isolated for IFN-γ intracellular staining and CD8α cell surface staining after ex vivo re-stimulation with OVA257-264 peptide. Graph depicts pooled data ± s.e.m.. (b) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were adoptively transferred into Ly5.2+ WT or TL- recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. Donor OT-I cells were tracked in the spleen and IEL 2 months p.i. Graph depicts pooled data ± s.e.m.. (c) 1 × 106 naïve Ly5.1+ OT-I cells were transferred into WT or TL-Tg recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. The donor OT-I cells in the spleens and IEL were tracked two months p.i. Graph depicts pooled data ± s.e.m.. (d) 5 × 104 Ly5.1+ naïve OT-I cells were transferred into C57BL/6 mice which were subsequently immunized i.v. with 5 × 105 OVAp-loaded DCs generated from bone marrow cells (BMDC) of WT or TL-Tg mice (top). Alternatively, OT-I cells were primed in vitro with TL negative APC or APC transfected with TL and then transferred to B6 recipients (bottom). Memory OT-I cells in the spleen were analyzed 2 months after DC immunization or after transfer of in vitro activated OT-I cells. Graph depicts pooled data ± s.e.m.. (e) Spleen or mLN DC were sorted based on the CD11c and MHC II expression and assessed for TL expression by flow cytometry. (f) Freshly isolated SPL or mLN DC were activated with CpG for 1 d and then analyzed for TL expression. * P < 0.05 and ** P < 0.001 (unpaired t-test). All data are representative of three independent experiments.

Mentions: Considering the class I-like antigen presenting molecules encoded by the mouse genome, TL is distinguished because it has a particularly high affinity for CD8αα, due to unique amino acids substitutions at three positions in the membrane proximal α3 domain17. To assess if TL, the most likely physiologic ligand for CD8αα in vivo12, also plays a role in the generation of CD8αβ effector memory cells, we analyzed CD8αβ T cell memory differentiation in TL gene (H2-T3b) knock out mice18, referred here as TL- mice. The absence of TL did not impair, but rather enhanced, the generation of OVA-specific CD8αβ memory T cells in the spleen of TL- mice infected orally with Lm bacteria expressing OVA antigen (Lm-OVA) (Fig. 1a). A similar effect was observed in the epithelium of the intestine when intraepithelial lymphocytes (IELs) were analyzed (Fig. 1a). Likewise, naïve OVA peptide (OVAp) and H-2Kb specific monoclonal TCR transgenic OT-I T cells transferred to WT or TL- recipient mice that were subsequently orally infected with Lm-OVA also generated more OT-I TCR transgenic memory T cells in the absence of TL expression (Fig. 1b). These observations are consistent with previous published data using single chain MHC class I transgenic mice on a β2m-deficient background, which also indicated that in the absence of TL, normal or slightly enhanced memory formed in response to a viral infection19. All together, the data show that, whereas CD8αα promotes memory differentiation of CD8αβ effector T cells12, its high-affinity ligand TL, appears to inhibit this process.


Mucosal memory CD8⁺ T cells are selected in the periphery by an MHC class I molecule.

Huang Y, Park Y, Wang-Zhu Y, Larange A, Arens R, Bernardo I, Olivares-Villagómez D, Herndler-Brandstetter D, Abraham N, Grubeck-Loebenstein B, Schoenberger SP, Van Kaer L, Kronenberg M, Teitell MA, Cheroutre H - Nat. Immunol. (2011)

TL negatively affects memory generation of CD8αβ+ T cells(a) WT or TL- mice were orally infected with 1 × 109 ActA- Lm-OVA. 30 days p.i., splenocytes (SPL) and IEL were isolated for IFN-γ intracellular staining and CD8α cell surface staining after ex vivo re-stimulation with OVA257-264 peptide. Graph depicts pooled data ± s.e.m.. (b) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were adoptively transferred into Ly5.2+ WT or TL- recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. Donor OT-I cells were tracked in the spleen and IEL 2 months p.i. Graph depicts pooled data ± s.e.m.. (c) 1 × 106 naïve Ly5.1+ OT-I cells were transferred into WT or TL-Tg recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. The donor OT-I cells in the spleens and IEL were tracked two months p.i. Graph depicts pooled data ± s.e.m.. (d) 5 × 104 Ly5.1+ naïve OT-I cells were transferred into C57BL/6 mice which were subsequently immunized i.v. with 5 × 105 OVAp-loaded DCs generated from bone marrow cells (BMDC) of WT or TL-Tg mice (top). Alternatively, OT-I cells were primed in vitro with TL negative APC or APC transfected with TL and then transferred to B6 recipients (bottom). Memory OT-I cells in the spleen were analyzed 2 months after DC immunization or after transfer of in vitro activated OT-I cells. Graph depicts pooled data ± s.e.m.. (e) Spleen or mLN DC were sorted based on the CD11c and MHC II expression and assessed for TL expression by flow cytometry. (f) Freshly isolated SPL or mLN DC were activated with CpG for 1 d and then analyzed for TL expression. * P < 0.05 and ** P < 0.001 (unpaired t-test). All data are representative of three independent experiments.
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Related In: Results  -  Collection

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Show All Figures
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Figure 1: TL negatively affects memory generation of CD8αβ+ T cells(a) WT or TL- mice were orally infected with 1 × 109 ActA- Lm-OVA. 30 days p.i., splenocytes (SPL) and IEL were isolated for IFN-γ intracellular staining and CD8α cell surface staining after ex vivo re-stimulation with OVA257-264 peptide. Graph depicts pooled data ± s.e.m.. (b) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were adoptively transferred into Ly5.2+ WT or TL- recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. Donor OT-I cells were tracked in the spleen and IEL 2 months p.i. Graph depicts pooled data ± s.e.m.. (c) 1 × 106 naïve Ly5.1+ OT-I cells were transferred into WT or TL-Tg recipients. One day after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. The donor OT-I cells in the spleens and IEL were tracked two months p.i. Graph depicts pooled data ± s.e.m.. (d) 5 × 104 Ly5.1+ naïve OT-I cells were transferred into C57BL/6 mice which were subsequently immunized i.v. with 5 × 105 OVAp-loaded DCs generated from bone marrow cells (BMDC) of WT or TL-Tg mice (top). Alternatively, OT-I cells were primed in vitro with TL negative APC or APC transfected with TL and then transferred to B6 recipients (bottom). Memory OT-I cells in the spleen were analyzed 2 months after DC immunization or after transfer of in vitro activated OT-I cells. Graph depicts pooled data ± s.e.m.. (e) Spleen or mLN DC were sorted based on the CD11c and MHC II expression and assessed for TL expression by flow cytometry. (f) Freshly isolated SPL or mLN DC were activated with CpG for 1 d and then analyzed for TL expression. * P < 0.05 and ** P < 0.001 (unpaired t-test). All data are representative of three independent experiments.
Mentions: Considering the class I-like antigen presenting molecules encoded by the mouse genome, TL is distinguished because it has a particularly high affinity for CD8αα, due to unique amino acids substitutions at three positions in the membrane proximal α3 domain17. To assess if TL, the most likely physiologic ligand for CD8αα in vivo12, also plays a role in the generation of CD8αβ effector memory cells, we analyzed CD8αβ T cell memory differentiation in TL gene (H2-T3b) knock out mice18, referred here as TL- mice. The absence of TL did not impair, but rather enhanced, the generation of OVA-specific CD8αβ memory T cells in the spleen of TL- mice infected orally with Lm bacteria expressing OVA antigen (Lm-OVA) (Fig. 1a). A similar effect was observed in the epithelium of the intestine when intraepithelial lymphocytes (IELs) were analyzed (Fig. 1a). Likewise, naïve OVA peptide (OVAp) and H-2Kb specific monoclonal TCR transgenic OT-I T cells transferred to WT or TL- recipient mice that were subsequently orally infected with Lm-OVA also generated more OT-I TCR transgenic memory T cells in the absence of TL expression (Fig. 1b). These observations are consistent with previous published data using single chain MHC class I transgenic mice on a β2m-deficient background, which also indicated that in the absence of TL, normal or slightly enhanced memory formed in response to a viral infection19. All together, the data show that, whereas CD8αα promotes memory differentiation of CD8αβ effector T cells12, its high-affinity ligand TL, appears to inhibit this process.

Bottom Line: The presence of immune memory at pathogen-entry sites is a prerequisite for protection.Nevertheless, the mechanisms that warrant immunity at peripheral interfaces are not understood.Furthermore, constitutive expression of TL on epithelial cells led to continued selection of mature CD8αβ(+) memory T cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Immunology, La Jolla Institute for Allergy & Immunology, La Jolla, California, USA.

ABSTRACT
The presence of immune memory at pathogen-entry sites is a prerequisite for protection. Nevertheless, the mechanisms that warrant immunity at peripheral interfaces are not understood. Here we show that the nonclassical major histocompatibility complex (MHC) class I molecule thymus leukemia antigen (TL), induced on dendritic cells interacting with CD8αα on activated CD8αβ(+) T cells, mediated affinity-based selection of memory precursor cells. Furthermore, constitutive expression of TL on epithelial cells led to continued selection of mature CD8αβ(+) memory T cells. The memory process driven by TL and CD8αα was essential for the generation of CD8αβ(+) memory T cells in the intestine and the accumulation of highly antigen-sensitive CD8αβ(+) memory T cells that form the first line of defense at the largest entry port for pathogens.

Show MeSH
Related in: MedlinePlus