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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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CD8αα expression correlates with the intensity of TCR activation(a) Total splenocytes were cultured in the presence of graded concentration of soluble anti-CD3 and anti-CD28. CD8αα expression, as measured by TL tetramer staining, was analyzed 3 d after in vitro culture. Representative data on gated CD8+ T cells are shown. IL-7Rα expression is also depicted. Three independent experiments were performed. (b) Naïve OT-I cells were cultured with artificial APC (MEC.B7) in the presence of graded concentration of OVA257-264 SIINFEKL (N4) or altered peptide ligands (Q4R7 and Q4). CD8αα expression was detected 2 d after in vitro culture. Three independent experiments were performed. (c) 1 x105 or 1 × 103 sorted naïve Ly5.1+ CD8+ OT-I cells were transferred into B6 recipient mice. 1 d after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. 7 d p.i., CD8αα expression was analyzed on Ly5.1+ CD8+ OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 4 mice per group). (d) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were transferred into WT recipient mice. 1 d after transfer, mice were orally infected with 2 × 108 WT Lm-Q4OVA or Lm-N4OVA. 7 d p.i., CD8αα expression was analyzed on donor OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 5 mice per group). Data are representative of three (a, b) and two (c, d) independent experiments.
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Figure 4: CD8αα expression correlates with the intensity of TCR activation(a) Total splenocytes were cultured in the presence of graded concentration of soluble anti-CD3 and anti-CD28. CD8αα expression, as measured by TL tetramer staining, was analyzed 3 d after in vitro culture. Representative data on gated CD8+ T cells are shown. IL-7Rα expression is also depicted. Three independent experiments were performed. (b) Naïve OT-I cells were cultured with artificial APC (MEC.B7) in the presence of graded concentration of OVA257-264 SIINFEKL (N4) or altered peptide ligands (Q4R7 and Q4). CD8αα expression was detected 2 d after in vitro culture. Three independent experiments were performed. (c) 1 x105 or 1 × 103 sorted naïve Ly5.1+ CD8+ OT-I cells were transferred into B6 recipient mice. 1 d after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. 7 d p.i., CD8αα expression was analyzed on Ly5.1+ CD8+ OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 4 mice per group). (d) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were transferred into WT recipient mice. 1 d after transfer, mice were orally infected with 2 × 108 WT Lm-Q4OVA or Lm-N4OVA. 7 d p.i., CD8αα expression was analyzed on donor OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 5 mice per group). Data are representative of three (a, b) and two (c, d) independent experiments.

Mentions: Due to the relatively high affinity interaction of CD8αα with TL, activation-induced expression of CD8αα can be distinguished from CD8αβ co-receptor expression using TL tetramers12. Activated CD8αβ T cells do not all induce CD8αα to the same extent and a variegated expression pattern of high (CD8ααhi) and low (CD8ααlo/-) expression is typically observed using TL tetramers. Together with the fact that the initial induction of CD8αα requires TCR stimulation, this finding suggests that there might be a close link between the intensity of TCR activation and the degree of CD8αα induction. In support of this notion, TL tetramer staining of polyclonal CD8αβ+ wild-type T lymphocytes activated in vitro with variable concentrations of soluble anti-CD3 and anti-CD28 showed graded increase in CD8αα expression with higher concentrations of CD3 plus CD28 mAbs (Fig. 4a). Furthermore, OT-I T cells stimulated with different altered peptide ligands (APLs) that bind equally well to the Kb class I molecule as the original OT-I ligand, SIINFEKL (N4), but which have different antigenic potencies31, also showed a tight association between the extent of CD8αα induction and the degree of TCR activation. Therefore the high affinity N4 ligand induced the highest amounts of CD8αα expression compared to the lower-affinity altered peptide ligands (Fig. 4b). Similar results of affinity-based induction of CD8αα were obtained in vivo when mice were analyzed that were infected orally with Lm-OVA after they received either a high- or a low-precursor frequency of naïve donor OT-I cells, which because of antigenic competition will model a low or high antigen dose, respectively (Fig. 4c). Additionally, we analyzed recipient mice adoptively transferred with equal amounts of OT-I precursor cells, but orally infected with bacteria expressing either the low-affinity Q4 (Lm-Q4)- or the high-affinity N4 (Lm-N4)-antigen. Mice receiving higher affinity N4 as compared to Q4 antigen generated more CD8αα-expressing CD8αβ T cells (Fig. 4d). In each of these approaches, the results consistently indicated that the extent of CD8αα induction represents a sensitive measurement for the intensity of the signal strength received through the activated TCR.


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)

CD8αα expression correlates with the intensity of TCR activation(a) Total splenocytes were cultured in the presence of graded concentration of soluble anti-CD3 and anti-CD28. CD8αα expression, as measured by TL tetramer staining, was analyzed 3 d after in vitro culture. Representative data on gated CD8+ T cells are shown. IL-7Rα expression is also depicted. Three independent experiments were performed. (b) Naïve OT-I cells were cultured with artificial APC (MEC.B7) in the presence of graded concentration of OVA257-264 SIINFEKL (N4) or altered peptide ligands (Q4R7 and Q4). CD8αα expression was detected 2 d after in vitro culture. Three independent experiments were performed. (c) 1 x105 or 1 × 103 sorted naïve Ly5.1+ CD8+ OT-I cells were transferred into B6 recipient mice. 1 d after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. 7 d p.i., CD8αα expression was analyzed on Ly5.1+ CD8+ OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 4 mice per group). (d) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were transferred into WT recipient mice. 1 d after transfer, mice were orally infected with 2 × 108 WT Lm-Q4OVA or Lm-N4OVA. 7 d p.i., CD8αα expression was analyzed on donor OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 5 mice per group). Data are representative of three (a, b) and two (c, d) independent experiments.
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Figure 4: CD8αα expression correlates with the intensity of TCR activation(a) Total splenocytes were cultured in the presence of graded concentration of soluble anti-CD3 and anti-CD28. CD8αα expression, as measured by TL tetramer staining, was analyzed 3 d after in vitro culture. Representative data on gated CD8+ T cells are shown. IL-7Rα expression is also depicted. Three independent experiments were performed. (b) Naïve OT-I cells were cultured with artificial APC (MEC.B7) in the presence of graded concentration of OVA257-264 SIINFEKL (N4) or altered peptide ligands (Q4R7 and Q4). CD8αα expression was detected 2 d after in vitro culture. Three independent experiments were performed. (c) 1 x105 or 1 × 103 sorted naïve Ly5.1+ CD8+ OT-I cells were transferred into B6 recipient mice. 1 d after transfer, mice were orally infected with 1 × 109 ActA- Lm-OVA. 7 d p.i., CD8αα expression was analyzed on Ly5.1+ CD8+ OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 4 mice per group). (d) 5 × 104 naïve Ly5.1+ CD8+ OT-I cells were transferred into WT recipient mice. 1 d after transfer, mice were orally infected with 2 × 108 WT Lm-Q4OVA or Lm-N4OVA. 7 d p.i., CD8αα expression was analyzed on donor OT-I cells from the spleen and IEL (representative data from a single mouse is shown, n = 5 mice per group). Data are representative of three (a, b) and two (c, d) independent experiments.
Mentions: Due to the relatively high affinity interaction of CD8αα with TL, activation-induced expression of CD8αα can be distinguished from CD8αβ co-receptor expression using TL tetramers12. Activated CD8αβ T cells do not all induce CD8αα to the same extent and a variegated expression pattern of high (CD8ααhi) and low (CD8ααlo/-) expression is typically observed using TL tetramers. Together with the fact that the initial induction of CD8αα requires TCR stimulation, this finding suggests that there might be a close link between the intensity of TCR activation and the degree of CD8αα induction. In support of this notion, TL tetramer staining of polyclonal CD8αβ+ wild-type T lymphocytes activated in vitro with variable concentrations of soluble anti-CD3 and anti-CD28 showed graded increase in CD8αα expression with higher concentrations of CD3 plus CD28 mAbs (Fig. 4a). Furthermore, OT-I T cells stimulated with different altered peptide ligands (APLs) that bind equally well to the Kb class I molecule as the original OT-I ligand, SIINFEKL (N4), but which have different antigenic potencies31, also showed a tight association between the extent of CD8αα induction and the degree of TCR activation. Therefore the high affinity N4 ligand induced the highest amounts of CD8αα expression compared to the lower-affinity altered peptide ligands (Fig. 4b). Similar results of affinity-based induction of CD8αα were obtained in vivo when mice were analyzed that were infected orally with Lm-OVA after they received either a high- or a low-precursor frequency of naïve donor OT-I cells, which because of antigenic competition will model a low or high antigen dose, respectively (Fig. 4c). Additionally, we analyzed recipient mice adoptively transferred with equal amounts of OT-I precursor cells, but orally infected with bacteria expressing either the low-affinity Q4 (Lm-Q4)- or the high-affinity N4 (Lm-N4)-antigen. Mice receiving higher affinity N4 as compared to Q4 antigen generated more CD8αα-expressing CD8αβ T cells (Fig. 4d). In each of these approaches, the results consistently indicated that the extent of CD8αα induction represents a sensitive measurement for the intensity of the signal strength received through the activated TCR.

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