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In vivo instruction of suppressor commitment in naive T cells.

Apostolou I, von Boehmer H - J. Exp. Med. (2004)

Bottom Line: The induction of antigen-specific tolerance in the mature immune system of the intact organism has met with limited success.Therefore, nonspecific immunosuppression has been the treatment of choice to prevent unwanted immunity.Here, it is shown that prolonged subcutaneous infusion of low doses of peptide by means of osmotic pumps transforms mature T cells into CD4+25+ suppressor cells that can persist for long periods of time in the absence of antigen and confer specific immunologic tolerance upon challenge with antigen.

View Article: PubMed Central - PubMed

Affiliation: Dana-Farber Cancer Institute, Harvard Medical School, 44 Binney St., Boston, MA 02115, USA.

ABSTRACT
The induction of antigen-specific tolerance in the mature immune system of the intact organism has met with limited success. Therefore, nonspecific immunosuppression has been the treatment of choice to prevent unwanted immunity. Here, it is shown that prolonged subcutaneous infusion of low doses of peptide by means of osmotic pumps transforms mature T cells into CD4+25+ suppressor cells that can persist for long periods of time in the absence of antigen and confer specific immunologic tolerance upon challenge with antigen. The described procedure resembles approaches of tolerance induction used decades ago, induces tolerance in the absence of immunity, and holds the promise to become an effective means of inducing antigen-specific tolerance prospectively, whereas its power to suppress already ongoing immune responses remains to be determined.

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Peptide dose–response relationship. (A) Percentages of 6.5+CD4+ (left) and of CD25+6.5+CD4+ (right) splenocytes in mice given either PBS (10 μg HA peptide/d) or 10−3, 10−1, 1, and 10 μg HA peptide/d for 14 d. (B) Expression of various markers by 6.5+CD4+ T cells of control mice (shaded histograms) and CD25+ and CD25− subsets of 6.5+CD4+ cells (unshaded histograms) of mice 14 d after implantation of osmotic pumps delivering 10−1 μg HA peptide/d. (C) Suppressive activity of CD25+CD4+ TCR-HA transgenic T cells from mice infused with various doses of HA peptide. Naive 6.5+CD4+ T cells from untreated TCR-HA,RAG-2−/− mice (N) were cultured alone or cocultured with sorted CD25+6.5+CD4+ T cells from Tx TCR-HA,RAG-2−/− mice (P, CD25+) given the indicated amount of peptide for 14 d in the presence of HA peptide-pulsed x-irradiated BALB/c nude splenocytes. Data are expressed as cpm ([3H]TdR incorporation) of coculture experiments over that of cultures of naive 6.5+CD4+ T cells from control TCR-HA,RAG-2−/− mice. One representative FACS® analysis of two independent experiments is shown for each peptide dosage in (A and B). All mice used in that experiment were age matched, Tx, and pump implanted at the same time.
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fig2: Peptide dose–response relationship. (A) Percentages of 6.5+CD4+ (left) and of CD25+6.5+CD4+ (right) splenocytes in mice given either PBS (10 μg HA peptide/d) or 10−3, 10−1, 1, and 10 μg HA peptide/d for 14 d. (B) Expression of various markers by 6.5+CD4+ T cells of control mice (shaded histograms) and CD25+ and CD25− subsets of 6.5+CD4+ cells (unshaded histograms) of mice 14 d after implantation of osmotic pumps delivering 10−1 μg HA peptide/d. (C) Suppressive activity of CD25+CD4+ TCR-HA transgenic T cells from mice infused with various doses of HA peptide. Naive 6.5+CD4+ T cells from untreated TCR-HA,RAG-2−/− mice (N) were cultured alone or cocultured with sorted CD25+6.5+CD4+ T cells from Tx TCR-HA,RAG-2−/− mice (P, CD25+) given the indicated amount of peptide for 14 d in the presence of HA peptide-pulsed x-irradiated BALB/c nude splenocytes. Data are expressed as cpm ([3H]TdR incorporation) of coculture experiments over that of cultures of naive 6.5+CD4+ T cells from control TCR-HA,RAG-2−/− mice. One representative FACS® analysis of two independent experiments is shown for each peptide dosage in (A and B). All mice used in that experiment were age matched, Tx, and pump implanted at the same time.

Mentions: Titration of the constantly supplied peptide dose indicated that a daily dose of as little as 0.001 μg was sufficient to induce an increase in the frequency of CD4+25+ cells (Fig. 2 A) that, however, never exceeded 25% of all CD4+ 6.5+ cells. When CD25+ and CD25− CD4+ T cells from pump-implanted mice were analyzed in vitro, the former but not the latter were anergic in terms of Ag-induced proliferation (Fig. 1 D) and could suppress the proliferative response of naive T cells in the standard in vitro assay that is often used to identify suppressor T cells (28), irrespective of the inducing peptide dose (Fig. 2 C and not depicted). Thus, in this assay, the unfractionated CD4+6.5+ population of cells was unresponsive to antigenic stimulation after peptide infusion (i.e., it was composed of suppressor and suppressed cells; Fig. 1 D).


In vivo instruction of suppressor commitment in naive T cells.

Apostolou I, von Boehmer H - J. Exp. Med. (2004)

Peptide dose–response relationship. (A) Percentages of 6.5+CD4+ (left) and of CD25+6.5+CD4+ (right) splenocytes in mice given either PBS (10 μg HA peptide/d) or 10−3, 10−1, 1, and 10 μg HA peptide/d for 14 d. (B) Expression of various markers by 6.5+CD4+ T cells of control mice (shaded histograms) and CD25+ and CD25− subsets of 6.5+CD4+ cells (unshaded histograms) of mice 14 d after implantation of osmotic pumps delivering 10−1 μg HA peptide/d. (C) Suppressive activity of CD25+CD4+ TCR-HA transgenic T cells from mice infused with various doses of HA peptide. Naive 6.5+CD4+ T cells from untreated TCR-HA,RAG-2−/− mice (N) were cultured alone or cocultured with sorted CD25+6.5+CD4+ T cells from Tx TCR-HA,RAG-2−/− mice (P, CD25+) given the indicated amount of peptide for 14 d in the presence of HA peptide-pulsed x-irradiated BALB/c nude splenocytes. Data are expressed as cpm ([3H]TdR incorporation) of coculture experiments over that of cultures of naive 6.5+CD4+ T cells from control TCR-HA,RAG-2−/− mice. One representative FACS® analysis of two independent experiments is shown for each peptide dosage in (A and B). All mice used in that experiment were age matched, Tx, and pump implanted at the same time.
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Related In: Results  -  Collection

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fig2: Peptide dose–response relationship. (A) Percentages of 6.5+CD4+ (left) and of CD25+6.5+CD4+ (right) splenocytes in mice given either PBS (10 μg HA peptide/d) or 10−3, 10−1, 1, and 10 μg HA peptide/d for 14 d. (B) Expression of various markers by 6.5+CD4+ T cells of control mice (shaded histograms) and CD25+ and CD25− subsets of 6.5+CD4+ cells (unshaded histograms) of mice 14 d after implantation of osmotic pumps delivering 10−1 μg HA peptide/d. (C) Suppressive activity of CD25+CD4+ TCR-HA transgenic T cells from mice infused with various doses of HA peptide. Naive 6.5+CD4+ T cells from untreated TCR-HA,RAG-2−/− mice (N) were cultured alone or cocultured with sorted CD25+6.5+CD4+ T cells from Tx TCR-HA,RAG-2−/− mice (P, CD25+) given the indicated amount of peptide for 14 d in the presence of HA peptide-pulsed x-irradiated BALB/c nude splenocytes. Data are expressed as cpm ([3H]TdR incorporation) of coculture experiments over that of cultures of naive 6.5+CD4+ T cells from control TCR-HA,RAG-2−/− mice. One representative FACS® analysis of two independent experiments is shown for each peptide dosage in (A and B). All mice used in that experiment were age matched, Tx, and pump implanted at the same time.
Mentions: Titration of the constantly supplied peptide dose indicated that a daily dose of as little as 0.001 μg was sufficient to induce an increase in the frequency of CD4+25+ cells (Fig. 2 A) that, however, never exceeded 25% of all CD4+ 6.5+ cells. When CD25+ and CD25− CD4+ T cells from pump-implanted mice were analyzed in vitro, the former but not the latter were anergic in terms of Ag-induced proliferation (Fig. 1 D) and could suppress the proliferative response of naive T cells in the standard in vitro assay that is often used to identify suppressor T cells (28), irrespective of the inducing peptide dose (Fig. 2 C and not depicted). Thus, in this assay, the unfractionated CD4+6.5+ population of cells was unresponsive to antigenic stimulation after peptide infusion (i.e., it was composed of suppressor and suppressed cells; Fig. 1 D).

Bottom Line: The induction of antigen-specific tolerance in the mature immune system of the intact organism has met with limited success.Therefore, nonspecific immunosuppression has been the treatment of choice to prevent unwanted immunity.Here, it is shown that prolonged subcutaneous infusion of low doses of peptide by means of osmotic pumps transforms mature T cells into CD4+25+ suppressor cells that can persist for long periods of time in the absence of antigen and confer specific immunologic tolerance upon challenge with antigen.

View Article: PubMed Central - PubMed

Affiliation: Dana-Farber Cancer Institute, Harvard Medical School, 44 Binney St., Boston, MA 02115, USA.

ABSTRACT
The induction of antigen-specific tolerance in the mature immune system of the intact organism has met with limited success. Therefore, nonspecific immunosuppression has been the treatment of choice to prevent unwanted immunity. Here, it is shown that prolonged subcutaneous infusion of low doses of peptide by means of osmotic pumps transforms mature T cells into CD4+25+ suppressor cells that can persist for long periods of time in the absence of antigen and confer specific immunologic tolerance upon challenge with antigen. The described procedure resembles approaches of tolerance induction used decades ago, induces tolerance in the absence of immunity, and holds the promise to become an effective means of inducing antigen-specific tolerance prospectively, whereas its power to suppress already ongoing immune responses remains to be determined.

Show MeSH
Related in: MedlinePlus