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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 induction of Th1 and Th2 type cells in vitro. Lymph node cells from an MBP-specific TCR transgenic mouse were stained with CD8,  B220, Mac-1, CD69, and CD44, and the negative cells were collected by flow cytometry as naive, CD4+ T cells (reanalysis of cells by flow cytometry indicated a population >98% CD4+). These cells were cultured in vitro at a concentration of 105 CD4+ cells and 106 irradiated nontransgenic syngeneic  splenocytes per well in 24-well plates with varying concentrations of Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. 10 d later, cells were washed several times and  restimulated at a concentration of 5 × 104 T cells plus 3 × 105 irradiated splenocytes per well in 96-well round-bottom plates with 10 μM Ac1-11, Ac111[4A], or Ac1-11[4Y]. Supernatants were collected at 48 h, and cytokines were detected by ELISA. Black bars, cytokine response to Ac1-11 in secondary challenge; shaded bars, cytokine response to Ac1-11[4A] in secondary challenge; cross-hatched bars, cytokine response to Ac1-11[4Y] in secondary  challenge. In general, Ac1-11 induced Th1 type responses regardless of primary concentration, Ac1-11[4A] induced Th2 type responses at high concentrations and Th1 type responses at low concentrations, and Ac1-11[4Y] induced primarily Th2 type responses, except at the lowest two doses. nd, not determined. Previous experiments indicated that too few live cells were recovered at the doses for which cytokine levels were not determined to test a secondary response.
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Figure 7: The induction of Th1 and Th2 type cells in vitro. Lymph node cells from an MBP-specific TCR transgenic mouse were stained with CD8, B220, Mac-1, CD69, and CD44, and the negative cells were collected by flow cytometry as naive, CD4+ T cells (reanalysis of cells by flow cytometry indicated a population >98% CD4+). These cells were cultured in vitro at a concentration of 105 CD4+ cells and 106 irradiated nontransgenic syngeneic splenocytes per well in 24-well plates with varying concentrations of Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. 10 d later, cells were washed several times and restimulated at a concentration of 5 × 104 T cells plus 3 × 105 irradiated splenocytes per well in 96-well round-bottom plates with 10 μM Ac1-11, Ac111[4A], or Ac1-11[4Y]. Supernatants were collected at 48 h, and cytokines were detected by ELISA. Black bars, cytokine response to Ac1-11 in secondary challenge; shaded bars, cytokine response to Ac1-11[4A] in secondary challenge; cross-hatched bars, cytokine response to Ac1-11[4Y] in secondary challenge. In general, Ac1-11 induced Th1 type responses regardless of primary concentration, Ac1-11[4A] induced Th2 type responses at high concentrations and Th1 type responses at low concentrations, and Ac1-11[4Y] induced primarily Th2 type responses, except at the lowest two doses. nd, not determined. Previous experiments indicated that too few live cells were recovered at the doses for which cytokine levels were not determined to test a secondary response.

Mentions: To ascertain whether Ac1-11 and Ac1-11 [4A] uniformly induce Th1 differentiation, and whether Ac1-11[4Y] uniformly induces Th2 differentiation, the cytokine profiles of purified naive CD4+ T cells in response to a secondary stimulation after a primary stimulation in vitro with various doses of either Ac1-11, Ac1-11[4A], or Ac1-11[4Y] were determined. Ac1-11 induced primarily Th1 type responses, whereas Ac1-11[4A] and Ac1-11[4Y] induced both Th1 and Th2 type responses, depending on the dose of peptide in the primary stimulation (Fig. 7). IL-2 production was inversely correlated with the concentration of peptide in the primary stimulation. Both high and low concentrations of the three peptides induced the most IFN-γ, with the exception that Ac1-11[4Y], regardless of dose, did not induce a large IFN-γ response. IL-4 production was highest at intermediate concentrations, corresponding to those that induced the least amount of IFN-γ. IL-10 production of cells stimulated by Ac1-11 in the primary culture was detected only in cells activated by the highest concentration of Ac1-11. IL-10 production by cells stimulated by Ac1-11[4A] and Ac1-11[4Y] followed a dose curve–response similar to that of IL-4 production.


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 induction of Th1 and Th2 type cells in vitro. Lymph node cells from an MBP-specific TCR transgenic mouse were stained with CD8,  B220, Mac-1, CD69, and CD44, and the negative cells were collected by flow cytometry as naive, CD4+ T cells (reanalysis of cells by flow cytometry indicated a population >98% CD4+). These cells were cultured in vitro at a concentration of 105 CD4+ cells and 106 irradiated nontransgenic syngeneic  splenocytes per well in 24-well plates with varying concentrations of Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. 10 d later, cells were washed several times and  restimulated at a concentration of 5 × 104 T cells plus 3 × 105 irradiated splenocytes per well in 96-well round-bottom plates with 10 μM Ac1-11, Ac111[4A], or Ac1-11[4Y]. Supernatants were collected at 48 h, and cytokines were detected by ELISA. Black bars, cytokine response to Ac1-11 in secondary challenge; shaded bars, cytokine response to Ac1-11[4A] in secondary challenge; cross-hatched bars, cytokine response to Ac1-11[4Y] in secondary  challenge. In general, Ac1-11 induced Th1 type responses regardless of primary concentration, Ac1-11[4A] induced Th2 type responses at high concentrations and Th1 type responses at low concentrations, and Ac1-11[4Y] induced primarily Th2 type responses, except at the lowest two doses. nd, not determined. Previous experiments indicated that too few live cells were recovered at the doses for which cytokine levels were not determined to test a secondary response.
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Related In: Results  -  Collection

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Figure 7: The induction of Th1 and Th2 type cells in vitro. Lymph node cells from an MBP-specific TCR transgenic mouse were stained with CD8, B220, Mac-1, CD69, and CD44, and the negative cells were collected by flow cytometry as naive, CD4+ T cells (reanalysis of cells by flow cytometry indicated a population >98% CD4+). These cells were cultured in vitro at a concentration of 105 CD4+ cells and 106 irradiated nontransgenic syngeneic splenocytes per well in 24-well plates with varying concentrations of Ac1-11, Ac1-11[4A], or Ac1-11[4Y]. 10 d later, cells were washed several times and restimulated at a concentration of 5 × 104 T cells plus 3 × 105 irradiated splenocytes per well in 96-well round-bottom plates with 10 μM Ac1-11, Ac111[4A], or Ac1-11[4Y]. Supernatants were collected at 48 h, and cytokines were detected by ELISA. Black bars, cytokine response to Ac1-11 in secondary challenge; shaded bars, cytokine response to Ac1-11[4A] in secondary challenge; cross-hatched bars, cytokine response to Ac1-11[4Y] in secondary challenge. In general, Ac1-11 induced Th1 type responses regardless of primary concentration, Ac1-11[4A] induced Th2 type responses at high concentrations and Th1 type responses at low concentrations, and Ac1-11[4Y] induced primarily Th2 type responses, except at the lowest two doses. nd, not determined. Previous experiments indicated that too few live cells were recovered at the doses for which cytokine levels were not determined to test a secondary response.
Mentions: To ascertain whether Ac1-11 and Ac1-11 [4A] uniformly induce Th1 differentiation, and whether Ac1-11[4Y] uniformly induces Th2 differentiation, the cytokine profiles of purified naive CD4+ T cells in response to a secondary stimulation after a primary stimulation in vitro with various doses of either Ac1-11, Ac1-11[4A], or Ac1-11[4Y] were determined. Ac1-11 induced primarily Th1 type responses, whereas Ac1-11[4A] and Ac1-11[4Y] induced both Th1 and Th2 type responses, depending on the dose of peptide in the primary stimulation (Fig. 7). IL-2 production was inversely correlated with the concentration of peptide in the primary stimulation. Both high and low concentrations of the three peptides induced the most IFN-γ, with the exception that Ac1-11[4Y], regardless of dose, did not induce a large IFN-γ response. IL-4 production was highest at intermediate concentrations, corresponding to those that induced the least amount of IFN-γ. IL-10 production of cells stimulated by Ac1-11 in the primary culture was detected only in cells activated by the highest concentration of Ac1-11. IL-10 production by cells stimulated by Ac1-11[4A] and Ac1-11[4Y] followed a dose curve–response similar to that of IL-4 production.

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