Limits...
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.

Show MeSH

Related in: MedlinePlus

Constitutive expression of TL on intestinal epithelial cells mediates selection of mature memory CD8αβ T cells(a,b) Naïve Ly5.1+ CD8+OT-I cells were cultured in the presence of APC (MEC.B7.SigOVA). After 2 days' culture, CD8ααhi and CD8ααlo/- OT-I cells were sorted and cultured for 3 more days in vitro. Then 0.5 × 106 CD8ααhi or CD8ααlo/- cells were adoptively transferred into B6 recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. Donor Ly5.1+ OT-I cells were tracked in the spleen and IEL 3 d (a) and 5 d (b) p.i.. Representative data from 3-4 mice in each group are shown. At least five independent experiments were performed. (c) As shown in (a), secondary OT-I memory cells were assessed in the IEL 45 d p.i.. Representative data from three to four mice in each group are shown. At least three independent experiments were performed. (d) Sorted in vitro activated Ly5.1+ CD8ααlo/- OT-I cells were cultured for 3 d and 0.5 × 106 primary effector cells were transferred into WT or TL-recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. 4 months p.i., memory OT-I cells were tracked in the spleens and IEL. Pooled data ± s.e.m. are shown. At least two independent experiments were performed. (e, f) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were orally infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. (g, h) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were intravenously infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. Data are representative of three independent experiments(e, f, g, h). (i) Ly5.1 mice adoptively transferred with 5 × 104 naïve WT or ΔE8I OT-I cells were orally immunized with 1 × 109 ActA- Lm-OVA. Two months after immunization, mice were re-challenged orally with 1 × 1010 WT Lm-OVA. Bacterial loads in the livers were assessed day 3 p.i.. Pooled data ± s.e.m. are shown (n = 6). Representative data are shown of three independent experiments. * P < 0.05; NS: not significant (unpaired t-test).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3197978&req=5

Figure 7: Constitutive expression of TL on intestinal epithelial cells mediates selection of mature memory CD8αβ T cells(a,b) Naïve Ly5.1+ CD8+OT-I cells were cultured in the presence of APC (MEC.B7.SigOVA). After 2 days' culture, CD8ααhi and CD8ααlo/- OT-I cells were sorted and cultured for 3 more days in vitro. Then 0.5 × 106 CD8ααhi or CD8ααlo/- cells were adoptively transferred into B6 recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. Donor Ly5.1+ OT-I cells were tracked in the spleen and IEL 3 d (a) and 5 d (b) p.i.. Representative data from 3-4 mice in each group are shown. At least five independent experiments were performed. (c) As shown in (a), secondary OT-I memory cells were assessed in the IEL 45 d p.i.. Representative data from three to four mice in each group are shown. At least three independent experiments were performed. (d) Sorted in vitro activated Ly5.1+ CD8ααlo/- OT-I cells were cultured for 3 d and 0.5 × 106 primary effector cells were transferred into WT or TL-recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. 4 months p.i., memory OT-I cells were tracked in the spleens and IEL. Pooled data ± s.e.m. are shown. At least two independent experiments were performed. (e, f) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were orally infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. (g, h) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were intravenously infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. Data are representative of three independent experiments(e, f, g, h). (i) Ly5.1 mice adoptively transferred with 5 × 104 naïve WT or ΔE8I OT-I cells were orally immunized with 1 × 109 ActA- Lm-OVA. Two months after immunization, mice were re-challenged orally with 1 × 1010 WT Lm-OVA. Bacterial loads in the livers were assessed day 3 p.i.. Pooled data ± s.e.m. are shown (n = 6). Representative data are shown of three independent experiments. * P < 0.05; NS: not significant (unpaired t-test).

Mentions: The constitutive TL expression on intestinal epithelial cells15,16, suggests that TL might continuously shape the resident mucosal memory CD8αβ+ T cell population even after rechallenge. To investigate this possibility, we examined the fate of primary and secondary CD8ααhi- or CD8ααlo/--CD8αβ effector T cells in vivo. OT-I cells initially primed in vitro in the absence of TL, using TL negative APCs, were sorted into CD8ααhi- and CD8ααlo/--primary effector T cells and adoptively transferred to wild-type recipient mice. Both subsets of primary effector cells displayed a similar short-term homing capacity (Supplementary Fig. 5a), and both effector T cell types responded alike when tested in vitro for interferon-γ production (Supplementary Fig. 5b) or in vivo for cytotoxicity (Supplementary Fig. 5c). However, in response to an oral re-challenge with Lm-OVA, primary memory OT-I T cells derived from the CD8ααhi precursors expanded in the spleen and in the intestine of the wild-type recipient mice, whereas memory cells from the CD8ααlo/- effector pool were only detectable in the host spleen, but not in the intestine (Fig. 7a). These data indicate that only CD8ααhi primary memory cells can persist long-term as mucosal TEM in proximity of TL constitutively expressed on the epithelial cells. Upon re-challenge, activated memory cells in lymphoid tissues expand and migrate as secondary effector cells to non-lymphoid tissues, such as the epithelium of the gut35. In agreement with this, comparable numbers of effector cells derived from either CD8ααhi or CD8ααlo/- primary effector OT-I cells were present in the intestine 5 days after recall (Fig. 7b). Nevertheless, when analyzed 45 days later, progeny of CD8ααhi effector cells were present as secondary memory cells in both the spleen and the intestine, whereas the CD8ααlo/- secondary effector cells did not remain as secondary TEM in the intestine (Fig. 7c). In contrast, in vitro primed CD8ααlo/-CD8αβ+ OT-I effector cells accumulated efficiently as TEM within the gut epithelium of TL- recipient mice (Fig. 7d), indicating that the constitutive expression of TL continues to mediate selective pressure that prevents the accumulation of CD8ααlo/- primary and secondary effector cells as mucosal TEM. These results also specify that CD8ααlo/- effector cells are not intrinsically incapable of converting to mucosal TEM, but that under normal physiological conditions, only CD8ααhi primary effector cells form long-lived mucosal TEM in proximity to TL constitutively expressed on the epithelial cells.


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)

Constitutive expression of TL on intestinal epithelial cells mediates selection of mature memory CD8αβ T cells(a,b) Naïve Ly5.1+ CD8+OT-I cells were cultured in the presence of APC (MEC.B7.SigOVA). After 2 days' culture, CD8ααhi and CD8ααlo/- OT-I cells were sorted and cultured for 3 more days in vitro. Then 0.5 × 106 CD8ααhi or CD8ααlo/- cells were adoptively transferred into B6 recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. Donor Ly5.1+ OT-I cells were tracked in the spleen and IEL 3 d (a) and 5 d (b) p.i.. Representative data from 3-4 mice in each group are shown. At least five independent experiments were performed. (c) As shown in (a), secondary OT-I memory cells were assessed in the IEL 45 d p.i.. Representative data from three to four mice in each group are shown. At least three independent experiments were performed. (d) Sorted in vitro activated Ly5.1+ CD8ααlo/- OT-I cells were cultured for 3 d and 0.5 × 106 primary effector cells were transferred into WT or TL-recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. 4 months p.i., memory OT-I cells were tracked in the spleens and IEL. Pooled data ± s.e.m. are shown. At least two independent experiments were performed. (e, f) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were orally infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. (g, h) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were intravenously infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. Data are representative of three independent experiments(e, f, g, h). (i) Ly5.1 mice adoptively transferred with 5 × 104 naïve WT or ΔE8I OT-I cells were orally immunized with 1 × 109 ActA- Lm-OVA. Two months after immunization, mice were re-challenged orally with 1 × 1010 WT Lm-OVA. Bacterial loads in the livers were assessed day 3 p.i.. Pooled data ± s.e.m. are shown (n = 6). Representative data are shown of three independent experiments. * P < 0.05; NS: not significant (unpaired t-test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3197978&req=5

Figure 7: Constitutive expression of TL on intestinal epithelial cells mediates selection of mature memory CD8αβ T cells(a,b) Naïve Ly5.1+ CD8+OT-I cells were cultured in the presence of APC (MEC.B7.SigOVA). After 2 days' culture, CD8ααhi and CD8ααlo/- OT-I cells were sorted and cultured for 3 more days in vitro. Then 0.5 × 106 CD8ααhi or CD8ααlo/- cells were adoptively transferred into B6 recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. Donor Ly5.1+ OT-I cells were tracked in the spleen and IEL 3 d (a) and 5 d (b) p.i.. Representative data from 3-4 mice in each group are shown. At least five independent experiments were performed. (c) As shown in (a), secondary OT-I memory cells were assessed in the IEL 45 d p.i.. Representative data from three to four mice in each group are shown. At least three independent experiments were performed. (d) Sorted in vitro activated Ly5.1+ CD8ααlo/- OT-I cells were cultured for 3 d and 0.5 × 106 primary effector cells were transferred into WT or TL-recipients. One month after transfer, mice were orally infected with 5 × 108 ActA- Lm-OVA. 4 months p.i., memory OT-I cells were tracked in the spleens and IEL. Pooled data ± s.e.m. are shown. At least two independent experiments were performed. (e, f) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were orally infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. (g, h) 5 × 104 naïve CD8+ OT-I cells were transferred into Ly5.1+ WT or Ly5.1+ TL- recipient mice. 1 d after transfer, mice were intravenously infected with Lm-Q4OVA. Effector OT-I cells in the peripheral blood (7 d p.i.) and memory OT-I cells (2 m p.i.) in the spleen and IEL were analyzed. Pooled data ± s.e.m. are shown. Data are representative of three independent experiments(e, f, g, h). (i) Ly5.1 mice adoptively transferred with 5 × 104 naïve WT or ΔE8I OT-I cells were orally immunized with 1 × 109 ActA- Lm-OVA. Two months after immunization, mice were re-challenged orally with 1 × 1010 WT Lm-OVA. Bacterial loads in the livers were assessed day 3 p.i.. Pooled data ± s.e.m. are shown (n = 6). Representative data are shown of three independent experiments. * P < 0.05; NS: not significant (unpaired t-test).
Mentions: The constitutive TL expression on intestinal epithelial cells15,16, suggests that TL might continuously shape the resident mucosal memory CD8αβ+ T cell population even after rechallenge. To investigate this possibility, we examined the fate of primary and secondary CD8ααhi- or CD8ααlo/--CD8αβ effector T cells in vivo. OT-I cells initially primed in vitro in the absence of TL, using TL negative APCs, were sorted into CD8ααhi- and CD8ααlo/--primary effector T cells and adoptively transferred to wild-type recipient mice. Both subsets of primary effector cells displayed a similar short-term homing capacity (Supplementary Fig. 5a), and both effector T cell types responded alike when tested in vitro for interferon-γ production (Supplementary Fig. 5b) or in vivo for cytotoxicity (Supplementary Fig. 5c). However, in response to an oral re-challenge with Lm-OVA, primary memory OT-I T cells derived from the CD8ααhi precursors expanded in the spleen and in the intestine of the wild-type recipient mice, whereas memory cells from the CD8ααlo/- effector pool were only detectable in the host spleen, but not in the intestine (Fig. 7a). These data indicate that only CD8ααhi primary memory cells can persist long-term as mucosal TEM in proximity of TL constitutively expressed on the epithelial cells. Upon re-challenge, activated memory cells in lymphoid tissues expand and migrate as secondary effector cells to non-lymphoid tissues, such as the epithelium of the gut35. In agreement with this, comparable numbers of effector cells derived from either CD8ααhi or CD8ααlo/- primary effector OT-I cells were present in the intestine 5 days after recall (Fig. 7b). Nevertheless, when analyzed 45 days later, progeny of CD8ααhi effector cells were present as secondary memory cells in both the spleen and the intestine, whereas the CD8ααlo/- secondary effector cells did not remain as secondary TEM in the intestine (Fig. 7c). In contrast, in vitro primed CD8ααlo/-CD8αβ+ OT-I effector cells accumulated efficiently as TEM within the gut epithelium of TL- recipient mice (Fig. 7d), indicating that the constitutive expression of TL continues to mediate selective pressure that prevents the accumulation of CD8ααlo/- primary and secondary effector cells as mucosal TEM. These results also specify that CD8ααlo/- effector cells are not intrinsically incapable of converting to mucosal TEM, but that under normal physiological conditions, only CD8ααhi primary effector cells form long-lived mucosal TEM in proximity to TL constitutively expressed on the epithelial cells.

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