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Induction of apoptosis and T helper 2 (Th2) responses correlates with peptide affinity for the major histocompatibility complex in self-reactive T cell receptor transgenic mice.

Pearson CI, van Ewijk W, McDevitt HO - J. Exp. Med. (1997)

Bottom Line: Concomitantly, T cells apoptose in the periphery, the degree of which correlates with the affinity of the peptide for the MHC.In addition, a shift in the T helper phenotype of the surviving T cells occurs such that the low affinity peptide, Ac1-11, induces primarily a Th1 response, whereas the highest affinity peptide, Ac1-11[4Y], induces primarily a Th2 type response.These data show that both the nature and the presumed number of the peptide-MHC complexes formed during specific peptide therapy affect both the degree of peripheral programmed cell death as well as the outcome of the T helper subset response in vivo, leading to amelioration of disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Stanford University Medical Center, California 94305, USA.

ABSTRACT
Multiple sclerosis is an autoimmune disease thought to be mediated by CD4+ T helper cells (Th). Experimental autoimmune encephalomyelitis is a rodent model of multiple sclerosis and has been used extensively to explore a variety of immunotherapies using soluble protein or peptide antigens. The underlying mechanisms of such therapy have been attributed to induction of T cell anergy, a switch in Th1 to Th2 responses, or peripheral deletion of autoreactive T cells. In this study, we have developed transgenic mice expressing a T cell receptor (TCR) specific for the NH2-terminal peptide Ac1-11 of the autoantigen myelin basic protein to explore the mechanism of soluble peptide therapy. T cells from these mice are highly skewed toward the CD4 population and have an abnormal thymic architecture, a phenomenon found in other TCR transgenic mice that exhibit a highly skewed CD4/CD8 ratio. Soluble Ac1-11 or the analogues Ac 1-11 [4A] or Ac1-11[4Y] (which bind to the major histocompatibility complex [MHC] class II molecule I-Au with increasing affinities) given intravenously activates T cells, rendering cells hyperresponsive in vitro for at least two days after injection. Concomitantly, T cells apoptose in the periphery, the degree of which correlates with the affinity of the peptide for the MHC. In addition, a shift in the T helper phenotype of the surviving T cells occurs such that the low affinity peptide, Ac1-11, induces primarily a Th1 response, whereas the highest affinity peptide, Ac1-11[4Y], induces primarily a Th2 type response. These data show that both the nature and the presumed number of the peptide-MHC complexes formed during specific peptide therapy affect both the degree of peripheral programmed cell death as well as the outcome of the T helper subset response in vivo, leading to amelioration of disease.

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The effect of the addition of anti–class II antibody  (10-3.6) on in vitro differentiation of naive TCR transgenic  mice T cells by 50, 5, and 0.5  μM Ac1-11[4Y]. Black bars represent responses of T cells cultured with 12.5 μg/ml 10-3.6  and Ac1-11[4Y], shaded bars  those of T cells cultured with  3.125 μg/ml 10-3.6 and Ac111[4Y], and cross-hatched bars  those of T cells cultured with  Ac1-11[4Y] alone. Sorted naive T  cells were cultured in 24-well  plates with irradiated splenocytes as APCs with various amounts of 10-3.6 and Ac1-11[4Y]. IL-2 production in the primary stimulation was determined  on culture supernatant 48 h after initial stimulation with Ac1-11[4Y] and 10-3.6. After a total of 7 d, viable cells were then washed and cultured with 10  μM Ac1-11[4Y] or medium only in 96-well round-bottom plates at 5 × 104 cells/well and 5 × 105 APCs/well for 48 h. IFN-γ and IL-4 production in  the secondary stimulation was determined as described. (A) IL-2 production by naive T cells in the primary stimulation is decreased by addition of 10-3.6.  (B) IFN-γ production by T cells in the secondary stimulation does not change significantly by addition of 10-3.6. (C) IL-4 production by T cells in the  secondary stimulation increases 2–20-fold, depending on the initial concentration of Ac1-11[4Y].
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Figure 8: The effect of the addition of anti–class II antibody (10-3.6) on in vitro differentiation of naive TCR transgenic mice T cells by 50, 5, and 0.5 μM Ac1-11[4Y]. Black bars represent responses of T cells cultured with 12.5 μg/ml 10-3.6 and Ac1-11[4Y], shaded bars those of T cells cultured with 3.125 μg/ml 10-3.6 and Ac111[4Y], and cross-hatched bars those of T cells cultured with Ac1-11[4Y] alone. Sorted naive T cells were cultured in 24-well plates with irradiated splenocytes as APCs with various amounts of 10-3.6 and Ac1-11[4Y]. IL-2 production in the primary stimulation was determined on culture supernatant 48 h after initial stimulation with Ac1-11[4Y] and 10-3.6. After a total of 7 d, viable cells were then washed and cultured with 10 μM Ac1-11[4Y] or medium only in 96-well round-bottom plates at 5 × 104 cells/well and 5 × 105 APCs/well for 48 h. IFN-γ and IL-4 production in the secondary stimulation was determined as described. (A) IL-2 production by naive T cells in the primary stimulation is decreased by addition of 10-3.6. (B) IFN-γ production by T cells in the secondary stimulation does not change significantly by addition of 10-3.6. (C) IL-4 production by T cells in the secondary stimulation increases 2–20-fold, depending on the initial concentration of Ac1-11[4Y].

Mentions: The different abilities of each of the three peptides to induce Th1 and Th2 responses may stem from the individual affinity of the peptide–MHC for the TCR or from differing numbers of peptide–MHC ligands interacting with TCR at the site of APC–T cell contact. To differentiate between these possibilities, two different concentrations of anti–class II antibody (10-3.6) were added to three different doses of Ac1-11[4Y] in the primary stimulation. At 12.5 μg/ml, 10-3.6 reduced the primary IL-2 production by 30–50%, whereas 3.125 μg/ml 10-3.6 reduced the primary IL-2 production by 5–30% (Fig. 8 A). Interestingly, suppression of the primary response by anti–class II antibody at 12.5 μg/ml induced stronger Th2 responses in the secondary stimulation at all three primary doses of Ac1-11[4Y] than did addition of no antibody (Fig. 8, B and C). Addition of 3.125 μg/ml 103.6 in the primary stimulation correlated with about twofold increased production of both IFN-γ and IL-4 in the secondary stimulation but did not change the ratio of IFN-γ to IL-4. In contrast, addition of 3.125 μg/ml 10-3.6 to a primary stimulation of 50 μM Ac1-11 inhibited primary IL-2 production by 30% and correlated with increases in IFN-γ responses of 20-fold in the secondary stimulation, while addition of 12.5 μg/ml 10-3.6 inhibited primary IL-2 production by ∼75% and correlated with increases in IFN-γ production of >200-fold in the secondary stimulation (data not shown). The production of IL-4 was similar between 12.5 μg/ml, 3.125 μg/ml, and no antibody in the Ac1-11 primary stimulation. These data indicate that the difference in affinity between Ac1-11 and Ac1-11[4Y] accounts for their differential ability to induce Th1 or Th2 responses.


Induction of apoptosis and T helper 2 (Th2) responses correlates with peptide affinity for the major histocompatibility complex in self-reactive T cell receptor transgenic mice.

Pearson CI, van Ewijk W, McDevitt HO - J. Exp. Med. (1997)

The effect of the addition of anti–class II antibody  (10-3.6) on in vitro differentiation of naive TCR transgenic  mice T cells by 50, 5, and 0.5  μM Ac1-11[4Y]. Black bars represent responses of T cells cultured with 12.5 μg/ml 10-3.6  and Ac1-11[4Y], shaded bars  those of T cells cultured with  3.125 μg/ml 10-3.6 and Ac111[4Y], and cross-hatched bars  those of T cells cultured with  Ac1-11[4Y] alone. Sorted naive T  cells were cultured in 24-well  plates with irradiated splenocytes as APCs with various amounts of 10-3.6 and Ac1-11[4Y]. IL-2 production in the primary stimulation was determined  on culture supernatant 48 h after initial stimulation with Ac1-11[4Y] and 10-3.6. After a total of 7 d, viable cells were then washed and cultured with 10  μM Ac1-11[4Y] or medium only in 96-well round-bottom plates at 5 × 104 cells/well and 5 × 105 APCs/well for 48 h. IFN-γ and IL-4 production in  the secondary stimulation was determined as described. (A) IL-2 production by naive T cells in the primary stimulation is decreased by addition of 10-3.6.  (B) IFN-γ production by T cells in the secondary stimulation does not change significantly by addition of 10-3.6. (C) IL-4 production by T cells in the  secondary stimulation increases 2–20-fold, depending on the initial concentration of Ac1-11[4Y].
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Related In: Results  -  Collection

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Figure 8: The effect of the addition of anti–class II antibody (10-3.6) on in vitro differentiation of naive TCR transgenic mice T cells by 50, 5, and 0.5 μM Ac1-11[4Y]. Black bars represent responses of T cells cultured with 12.5 μg/ml 10-3.6 and Ac1-11[4Y], shaded bars those of T cells cultured with 3.125 μg/ml 10-3.6 and Ac111[4Y], and cross-hatched bars those of T cells cultured with Ac1-11[4Y] alone. Sorted naive T cells were cultured in 24-well plates with irradiated splenocytes as APCs with various amounts of 10-3.6 and Ac1-11[4Y]. IL-2 production in the primary stimulation was determined on culture supernatant 48 h after initial stimulation with Ac1-11[4Y] and 10-3.6. After a total of 7 d, viable cells were then washed and cultured with 10 μM Ac1-11[4Y] or medium only in 96-well round-bottom plates at 5 × 104 cells/well and 5 × 105 APCs/well for 48 h. IFN-γ and IL-4 production in the secondary stimulation was determined as described. (A) IL-2 production by naive T cells in the primary stimulation is decreased by addition of 10-3.6. (B) IFN-γ production by T cells in the secondary stimulation does not change significantly by addition of 10-3.6. (C) IL-4 production by T cells in the secondary stimulation increases 2–20-fold, depending on the initial concentration of Ac1-11[4Y].
Mentions: The different abilities of each of the three peptides to induce Th1 and Th2 responses may stem from the individual affinity of the peptide–MHC for the TCR or from differing numbers of peptide–MHC ligands interacting with TCR at the site of APC–T cell contact. To differentiate between these possibilities, two different concentrations of anti–class II antibody (10-3.6) were added to three different doses of Ac1-11[4Y] in the primary stimulation. At 12.5 μg/ml, 10-3.6 reduced the primary IL-2 production by 30–50%, whereas 3.125 μg/ml 10-3.6 reduced the primary IL-2 production by 5–30% (Fig. 8 A). Interestingly, suppression of the primary response by anti–class II antibody at 12.5 μg/ml induced stronger Th2 responses in the secondary stimulation at all three primary doses of Ac1-11[4Y] than did addition of no antibody (Fig. 8, B and C). Addition of 3.125 μg/ml 103.6 in the primary stimulation correlated with about twofold increased production of both IFN-γ and IL-4 in the secondary stimulation but did not change the ratio of IFN-γ to IL-4. In contrast, addition of 3.125 μg/ml 10-3.6 to a primary stimulation of 50 μM Ac1-11 inhibited primary IL-2 production by 30% and correlated with increases in IFN-γ responses of 20-fold in the secondary stimulation, while addition of 12.5 μg/ml 10-3.6 inhibited primary IL-2 production by ∼75% and correlated with increases in IFN-γ production of >200-fold in the secondary stimulation (data not shown). The production of IL-4 was similar between 12.5 μg/ml, 3.125 μg/ml, and no antibody in the Ac1-11 primary stimulation. These data indicate that the difference in affinity between Ac1-11 and Ac1-11[4Y] accounts for their differential ability to induce Th1 or Th2 responses.

Bottom Line: Concomitantly, T cells apoptose in the periphery, the degree of which correlates with the affinity of the peptide for the MHC.In addition, a shift in the T helper phenotype of the surviving T cells occurs such that the low affinity peptide, Ac1-11, induces primarily a Th1 response, whereas the highest affinity peptide, Ac1-11[4Y], induces primarily a Th2 type response.These data show that both the nature and the presumed number of the peptide-MHC complexes formed during specific peptide therapy affect both the degree of peripheral programmed cell death as well as the outcome of the T helper subset response in vivo, leading to amelioration of disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Stanford University Medical Center, California 94305, USA.

ABSTRACT
Multiple sclerosis is an autoimmune disease thought to be mediated by CD4+ T helper cells (Th). Experimental autoimmune encephalomyelitis is a rodent model of multiple sclerosis and has been used extensively to explore a variety of immunotherapies using soluble protein or peptide antigens. The underlying mechanisms of such therapy have been attributed to induction of T cell anergy, a switch in Th1 to Th2 responses, or peripheral deletion of autoreactive T cells. In this study, we have developed transgenic mice expressing a T cell receptor (TCR) specific for the NH2-terminal peptide Ac1-11 of the autoantigen myelin basic protein to explore the mechanism of soluble peptide therapy. T cells from these mice are highly skewed toward the CD4 population and have an abnormal thymic architecture, a phenomenon found in other TCR transgenic mice that exhibit a highly skewed CD4/CD8 ratio. Soluble Ac1-11 or the analogues Ac 1-11 [4A] or Ac1-11[4Y] (which bind to the major histocompatibility complex [MHC] class II molecule I-Au with increasing affinities) given intravenously activates T cells, rendering cells hyperresponsive in vitro for at least two days after injection. Concomitantly, T cells apoptose in the periphery, the degree of which correlates with the affinity of the peptide for the MHC. In addition, a shift in the T helper phenotype of the surviving T cells occurs such that the low affinity peptide, Ac1-11, induces primarily a Th1 response, whereas the highest affinity peptide, Ac1-11[4Y], induces primarily a Th2 type response. These data show that both the nature and the presumed number of the peptide-MHC complexes formed during specific peptide therapy affect both the degree of peripheral programmed cell death as well as the outcome of the T helper subset response in vivo, leading to amelioration of disease.

Show MeSH
Related in: MedlinePlus