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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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Cytokine profiles of splenocytes from mice injected with PBS, Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. Increasing  amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokine IFN-γ were detected depending on  the peptide injected. Cells were cultured at 3–5 × 105 cells per well in 96-well round-bottom plates with either medium only, Ac1-11, Ac1-11[4A], or  Ac1-11[4Y]. Supernatants were collected 45–52 h later, and the amount of IL-2, IFN-γ, IL-4, and IL-10 was determined by ELISA. Black bars, cytokine  production in response to Ac1-11 stimulation; shaded bars, cytokine production in response to Ac1-11[4A]; and cross-hatched bars, cytokine production  in response to Ac1-11[4Y]. (A) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after  administration. (B) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 6 d after administration. Similar to day 1, increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokines IL-2 and  IFN-γ were detected depending on the peptide injected. No cytokines were detected in supernatants from cells incubated with medium only.
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Figure 6: Cytokine profiles of splenocytes from mice injected with PBS, Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. Increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokine IFN-γ were detected depending on the peptide injected. Cells were cultured at 3–5 × 105 cells per well in 96-well round-bottom plates with either medium only, Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. Supernatants were collected 45–52 h later, and the amount of IL-2, IFN-γ, IL-4, and IL-10 was determined by ELISA. Black bars, cytokine production in response to Ac1-11 stimulation; shaded bars, cytokine production in response to Ac1-11[4A]; and cross-hatched bars, cytokine production in response to Ac1-11[4Y]. (A) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. (B) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 6 d after administration. Similar to day 1, increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokines IL-2 and IFN-γ were detected depending on the peptide injected. No cytokines were detected in supernatants from cells incubated with medium only.

Mentions: Since a large population of antigen-responsive T cells survive the single injection of Ac1-11, Ac1-11[4A], or Ac1-11[4Y], we examined whether these peptides differed in induction of T helper subsets. TCR transgenic spleen cells when cultured in vitro with 100 μM Ac1-11[4A] or Ac1-11[4Y] (the concentration at which T cells proliferate equally) produce >200 U/ml IL-2, <50 ng/ml IFN-γ, and little or no detectable IL-4 or IL-10, which is consistent with a Th0 profile (Fig. 6). Lymph node cells (105) cultured with irradiated syngeneic splenocytes (5 × 105) displayed similar cytokine profiles (data not shown). Within 24 h after injection of 2.4 mg of peptide, spleen or lymph node cells from Ac1-11– or Ac1-11[4A]–injected mice produce more IL-2 and up to 10 times more IFN-γ. Little or no IL-4 is detectable, and <1 ng/ml IL-10 is produced (Fig. 6 A). In contrast, cells from Ac1-11[4Y]–injected mice make the least amount of IL-2, IFN-γ at levels comparable to those produced by cells from naive mice, detectable levels of IL-4 (up to 1 ng/ml), and 2–6 times more IL-10 than that produced by cells from Ac1-11– and Ac1-11[4A]– injected mice (Fig. 6 A). By day 6, IFN-γ production was comparable between the different treatment groups. Cells from Ac1-11[4Y] consistently produce the least amount of IFN-γ. The amount of IL-4 or IL-10 produced increased with the affinity of the peptide for I-Au (Fig. 6 B). Thus, a single dose of low or medium affinity peptides, Ac1-11 and Ac1-11[4A], induces differentiation into the Th1 subset, while a single dose of a high affinity peptide, Ac1-11[4Y], induces differentiation into the Th2 subset. Similar results were obtained after a single dose of 240 μg of peptide (data not shown).


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)

Cytokine profiles of splenocytes from mice injected with PBS, Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. Increasing  amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokine IFN-γ were detected depending on  the peptide injected. Cells were cultured at 3–5 × 105 cells per well in 96-well round-bottom plates with either medium only, Ac1-11, Ac1-11[4A], or  Ac1-11[4Y]. Supernatants were collected 45–52 h later, and the amount of IL-2, IFN-γ, IL-4, and IL-10 was determined by ELISA. Black bars, cytokine  production in response to Ac1-11 stimulation; shaded bars, cytokine production in response to Ac1-11[4A]; and cross-hatched bars, cytokine production  in response to Ac1-11[4Y]. (A) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after  administration. (B) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 6 d after administration. Similar to day 1, increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokines IL-2 and  IFN-γ were detected depending on the peptide injected. No cytokines were detected in supernatants from cells incubated with medium only.
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Figure 6: Cytokine profiles of splenocytes from mice injected with PBS, Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. Increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokine IFN-γ were detected depending on the peptide injected. Cells were cultured at 3–5 × 105 cells per well in 96-well round-bottom plates with either medium only, Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. Supernatants were collected 45–52 h later, and the amount of IL-2, IFN-γ, IL-4, and IL-10 was determined by ELISA. Black bars, cytokine production in response to Ac1-11 stimulation; shaded bars, cytokine production in response to Ac1-11[4A]; and cross-hatched bars, cytokine production in response to Ac1-11[4Y]. (A) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 1 d after administration. (B) Cytokine profiles of splenocytes from mice injected with PBS or 2.4 mg of Ac1-11, Ac1-11[4A], or Ac1-11[4Y] 6 d after administration. Similar to day 1, increasing amounts of Th2 type cytokines IL-4 and IL-10 were detected, while decreasing amounts of the Th1 type cytokines IL-2 and IFN-γ were detected depending on the peptide injected. No cytokines were detected in supernatants from cells incubated with medium only.
Mentions: Since a large population of antigen-responsive T cells survive the single injection of Ac1-11, Ac1-11[4A], or Ac1-11[4Y], we examined whether these peptides differed in induction of T helper subsets. TCR transgenic spleen cells when cultured in vitro with 100 μM Ac1-11[4A] or Ac1-11[4Y] (the concentration at which T cells proliferate equally) produce >200 U/ml IL-2, <50 ng/ml IFN-γ, and little or no detectable IL-4 or IL-10, which is consistent with a Th0 profile (Fig. 6). Lymph node cells (105) cultured with irradiated syngeneic splenocytes (5 × 105) displayed similar cytokine profiles (data not shown). Within 24 h after injection of 2.4 mg of peptide, spleen or lymph node cells from Ac1-11– or Ac1-11[4A]–injected mice produce more IL-2 and up to 10 times more IFN-γ. Little or no IL-4 is detectable, and <1 ng/ml IL-10 is produced (Fig. 6 A). In contrast, cells from Ac1-11[4Y]–injected mice make the least amount of IL-2, IFN-γ at levels comparable to those produced by cells from naive mice, detectable levels of IL-4 (up to 1 ng/ml), and 2–6 times more IL-10 than that produced by cells from Ac1-11– and Ac1-11[4A]– injected mice (Fig. 6 A). By day 6, IFN-γ production was comparable between the different treatment groups. Cells from Ac1-11[4Y] consistently produce the least amount of IFN-γ. The amount of IL-4 or IL-10 produced increased with the affinity of the peptide for I-Au (Fig. 6 B). Thus, a single dose of low or medium affinity peptides, Ac1-11 and Ac1-11[4A], induces differentiation into the Th1 subset, while a single dose of a high affinity peptide, Ac1-11[4Y], induces differentiation into the Th2 subset. Similar results were obtained after a single dose of 240 μg of peptide (data not shown).

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