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Selective expansion of cross-reactive CD8(+) memory T cells by viral variants.

Haanen JB, Wolkers MC, Kruisbeek AM, Schumacher TN - J. Exp. Med. (1999)

Bottom Line: The role of memory T cells during the immune response against random antigenic variants has not been resolved.Here, we show by simultaneous staining with two tetrameric major histocompatibility complex (MHC)-peptide molecules, that the polyclonal CD8(+) T cell response against a series of natural variants of the influenza A nucleoprotein epitope is completely dominated by infrequent cross-reactive T cells that expand from an original memory population.These results provide direct evidence that the repertoire of antigen-specific T cells used during an infection critically depends on prior antigen encounters, and indicate that polyclonal memory T cell populations can provide protection against a range of antigenic variants.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, The Netherlands Cancer Institute, Amsterdam 1066 CX, The Netherlands.

ABSTRACT
The role of memory T cells during the immune response against random antigenic variants has not been resolved. Here, we show by simultaneous staining with two tetrameric major histocompatibility complex (MHC)-peptide molecules, that the polyclonal CD8(+) T cell response against a series of natural variants of the influenza A nucleoprotein epitope is completely dominated by infrequent cross-reactive T cells that expand from an original memory population. Based on both biochemical and functional criteria, these cross-reactive cytotoxic T cells productively recognize both the parental and the mutant epitope in vitro and in vivo. These results provide direct evidence that the repertoire of antigen-specific T cells used during an infection critically depends on prior antigen encounters, and indicate that polyclonal memory T cell populations can provide protection against a range of antigenic variants.

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Related in: MedlinePlus

Selective expansion of cross-reactive T cells. (A) Naive mice were infected intranasally with 10 hemagglutinin units (HAU) influenza A/NT/60/68 (H3N2; epitope: ASNENMDAM) or 200 HAU A/HKx31 (H3N2; epitope: ASNENMETM). 4–6 wk after the primary infection, mice were challenged with 200 HAU influenza A/NT/60/68 (panels 1 and 2) or A/PR/8/34 (panels 3 and 4) (H1N1; epitope: ASNENMETM). At day 10 after challenge, mice were killed and lung-infiltrating mononuclear cells were stained for CD8 and antigen-specific TCR expression using anti-CD8b mAb and MHC tetramers. Gated CD8+ T cells were analyzed for binding of APC-labeled MHC tetramers with the influenza A/NT/60/68 epitope and PE-labeled MHC tetramers with the influenza A/PR/8/34 epitope. Identical results are obtained when challenge is performed with influenza A 81/HO (H7N2; NP366–374 sequence ASNENMDAM) instead of A/NT/60/68, and is therefore independent of HN subtype (not shown). Results represent the outcome of more than five experiments performed. (B) Mice were sequentially infected with influenza virus A/HKx31 and A/NT/60/68. On day 7 after the second infection, spleen cells were incubated with MHC tetramers containing the NP366–374 epitope (ASNENMDAM) of A/NT/60/68. H-2Db monomers complexed with ASNENMDAM (▪) or ASNENMETM (▴) were added in increasing amounts, and inhibition of the binding of MHC tetramers to CD8+ T cells was determined.
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Figure 1: Selective expansion of cross-reactive T cells. (A) Naive mice were infected intranasally with 10 hemagglutinin units (HAU) influenza A/NT/60/68 (H3N2; epitope: ASNENMDAM) or 200 HAU A/HKx31 (H3N2; epitope: ASNENMETM). 4–6 wk after the primary infection, mice were challenged with 200 HAU influenza A/NT/60/68 (panels 1 and 2) or A/PR/8/34 (panels 3 and 4) (H1N1; epitope: ASNENMETM). At day 10 after challenge, mice were killed and lung-infiltrating mononuclear cells were stained for CD8 and antigen-specific TCR expression using anti-CD8b mAb and MHC tetramers. Gated CD8+ T cells were analyzed for binding of APC-labeled MHC tetramers with the influenza A/NT/60/68 epitope and PE-labeled MHC tetramers with the influenza A/PR/8/34 epitope. Identical results are obtained when challenge is performed with influenza A 81/HO (H7N2; NP366–374 sequence ASNENMDAM) instead of A/NT/60/68, and is therefore independent of HN subtype (not shown). Results represent the outcome of more than five experiments performed. (B) Mice were sequentially infected with influenza virus A/HKx31 and A/NT/60/68. On day 7 after the second infection, spleen cells were incubated with MHC tetramers containing the NP366–374 epitope (ASNENMDAM) of A/NT/60/68. H-2Db monomers complexed with ASNENMDAM (▪) or ASNENMETM (▴) were added in increasing amounts, and inhibition of the binding of MHC tetramers to CD8+ T cells was determined.

Mentions: To assess the effect of random antigen variation on the dynamics of T cell responses in vivo, we infected mice with pairs of influenza A viruses. These viruses expressed either the same NP366–374 epitope or epitope variants. The specificity of the resulting T cell repertoire was assessed by two-parameter MHC tetramer staining, and association of MHC tetramers to NK receptors was ruled out through analysis of CD8b-expressing cells only. When mice are infected once or twice with either influenza virus strain A/NT/60/68 or A/PR/8/34, which differ in the sequence of the immunodominant NP CTL epitope at positions 7 and 8 (ASNENMDAM vs. ASNENMETM), the vast majority of the resulting NP-specific T cells selectively recognize the epitope of the strain encountered and not that of the opposite strain (Fig. 1 A, panels 1 and 3). This dominant role of the peptide side chains at positions 7 and 8 in ligand recognition by the majority of T cells is in accord with the prominent contribution of p8 and especially p7 to the TCR-exposed surface of this peptide–MHC complex 22.


Selective expansion of cross-reactive CD8(+) memory T cells by viral variants.

Haanen JB, Wolkers MC, Kruisbeek AM, Schumacher TN - J. Exp. Med. (1999)

Selective expansion of cross-reactive T cells. (A) Naive mice were infected intranasally with 10 hemagglutinin units (HAU) influenza A/NT/60/68 (H3N2; epitope: ASNENMDAM) or 200 HAU A/HKx31 (H3N2; epitope: ASNENMETM). 4–6 wk after the primary infection, mice were challenged with 200 HAU influenza A/NT/60/68 (panels 1 and 2) or A/PR/8/34 (panels 3 and 4) (H1N1; epitope: ASNENMETM). At day 10 after challenge, mice were killed and lung-infiltrating mononuclear cells were stained for CD8 and antigen-specific TCR expression using anti-CD8b mAb and MHC tetramers. Gated CD8+ T cells were analyzed for binding of APC-labeled MHC tetramers with the influenza A/NT/60/68 epitope and PE-labeled MHC tetramers with the influenza A/PR/8/34 epitope. Identical results are obtained when challenge is performed with influenza A 81/HO (H7N2; NP366–374 sequence ASNENMDAM) instead of A/NT/60/68, and is therefore independent of HN subtype (not shown). Results represent the outcome of more than five experiments performed. (B) Mice were sequentially infected with influenza virus A/HKx31 and A/NT/60/68. On day 7 after the second infection, spleen cells were incubated with MHC tetramers containing the NP366–374 epitope (ASNENMDAM) of A/NT/60/68. H-2Db monomers complexed with ASNENMDAM (▪) or ASNENMETM (▴) were added in increasing amounts, and inhibition of the binding of MHC tetramers to CD8+ T cells was determined.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195685&req=5

Figure 1: Selective expansion of cross-reactive T cells. (A) Naive mice were infected intranasally with 10 hemagglutinin units (HAU) influenza A/NT/60/68 (H3N2; epitope: ASNENMDAM) or 200 HAU A/HKx31 (H3N2; epitope: ASNENMETM). 4–6 wk after the primary infection, mice were challenged with 200 HAU influenza A/NT/60/68 (panels 1 and 2) or A/PR/8/34 (panels 3 and 4) (H1N1; epitope: ASNENMETM). At day 10 after challenge, mice were killed and lung-infiltrating mononuclear cells were stained for CD8 and antigen-specific TCR expression using anti-CD8b mAb and MHC tetramers. Gated CD8+ T cells were analyzed for binding of APC-labeled MHC tetramers with the influenza A/NT/60/68 epitope and PE-labeled MHC tetramers with the influenza A/PR/8/34 epitope. Identical results are obtained when challenge is performed with influenza A 81/HO (H7N2; NP366–374 sequence ASNENMDAM) instead of A/NT/60/68, and is therefore independent of HN subtype (not shown). Results represent the outcome of more than five experiments performed. (B) Mice were sequentially infected with influenza virus A/HKx31 and A/NT/60/68. On day 7 after the second infection, spleen cells were incubated with MHC tetramers containing the NP366–374 epitope (ASNENMDAM) of A/NT/60/68. H-2Db monomers complexed with ASNENMDAM (▪) or ASNENMETM (▴) were added in increasing amounts, and inhibition of the binding of MHC tetramers to CD8+ T cells was determined.
Mentions: To assess the effect of random antigen variation on the dynamics of T cell responses in vivo, we infected mice with pairs of influenza A viruses. These viruses expressed either the same NP366–374 epitope or epitope variants. The specificity of the resulting T cell repertoire was assessed by two-parameter MHC tetramer staining, and association of MHC tetramers to NK receptors was ruled out through analysis of CD8b-expressing cells only. When mice are infected once or twice with either influenza virus strain A/NT/60/68 or A/PR/8/34, which differ in the sequence of the immunodominant NP CTL epitope at positions 7 and 8 (ASNENMDAM vs. ASNENMETM), the vast majority of the resulting NP-specific T cells selectively recognize the epitope of the strain encountered and not that of the opposite strain (Fig. 1 A, panels 1 and 3). This dominant role of the peptide side chains at positions 7 and 8 in ligand recognition by the majority of T cells is in accord with the prominent contribution of p8 and especially p7 to the TCR-exposed surface of this peptide–MHC complex 22.

Bottom Line: The role of memory T cells during the immune response against random antigenic variants has not been resolved.Here, we show by simultaneous staining with two tetrameric major histocompatibility complex (MHC)-peptide molecules, that the polyclonal CD8(+) T cell response against a series of natural variants of the influenza A nucleoprotein epitope is completely dominated by infrequent cross-reactive T cells that expand from an original memory population.These results provide direct evidence that the repertoire of antigen-specific T cells used during an infection critically depends on prior antigen encounters, and indicate that polyclonal memory T cell populations can provide protection against a range of antigenic variants.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, The Netherlands Cancer Institute, Amsterdam 1066 CX, The Netherlands.

ABSTRACT
The role of memory T cells during the immune response against random antigenic variants has not been resolved. Here, we show by simultaneous staining with two tetrameric major histocompatibility complex (MHC)-peptide molecules, that the polyclonal CD8(+) T cell response against a series of natural variants of the influenza A nucleoprotein epitope is completely dominated by infrequent cross-reactive T cells that expand from an original memory population. Based on both biochemical and functional criteria, these cross-reactive cytotoxic T cells productively recognize both the parental and the mutant epitope in vitro and in vivo. These results provide direct evidence that the repertoire of antigen-specific T cells used during an infection critically depends on prior antigen encounters, and indicate that polyclonal memory T cell populations can provide protection against a range of antigenic variants.

Show MeSH
Related in: MedlinePlus