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Transferable anergy: superantigen treatment induces CD4+ T cell tolerance that is reversible and requires CD4-CD8- cells and interferon gamma.

Cauley LS, Cauley KA, Shub F, Huston G, Swain SL - J. Exp. Med. (1997)

Bottom Line: Bacterial superantigens induce peripheral unresponsiveness in CD4+ T cell populations that express appropriate Vbeta chains.However, when the SEA-treated CD4+ cells were completely purified away from all other contaminating cells, they regained the ability to proliferate and secrete cytokines.Further analysis demonstrated that interferon gamma, but not the Fas receptor, played a critical role in the suppression.

View Article: PubMed Central - PubMed

Affiliation: Trudeau Institute, Saranac Lake, New York 12983, USA.

ABSTRACT
Bacterial superantigens induce peripheral unresponsiveness in CD4+ T cell populations that express appropriate Vbeta chains. We have used Vbeta3/Valpha11 T cell receptor transgenic (Tg) mice and the Vbeta3-specific superantigen staphylococcal enterotoxin A (SEA) to further investigate the mechanisms that contribute to such unresponsiveness. As in other models, in vivo exposure to SEA rendered the Tg CD4+ cells unresponsive to subsequent restimulation in vitro with antigen or mitogens. However, when the SEA-treated CD4+ cells were completely purified away from all other contaminating cells, they regained the ability to proliferate and secrete cytokines. Moreover, enriched CD4-CD8- cells from the SEA-treated mice suppressed the responses of fresh control CD4+ cells in mixed cultures indicating that the apparent "anergy" was both transferable and reversible. Further analysis demonstrated that interferon gamma, but not the Fas receptor, played a critical role in the suppression.

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Extensively purified CD4+ cells from SEA-treated mice can  synthesize cytokine mRNA. The SEA-mediated T cell anergy was reversed by FACS® sorting the CD4+ cells. (A) PCR analysis of cytokine  mRNAs from SEA-treated and control CD4+ T cells after high level purification by positive selection and restimulation with 2C11 and anti-CD28 Ab. PCR products were electrophoresed on 1.5% agarose gels  stained with ethidium bromide. Competitor DNA is not shown. (B)  PCR products were quantitated using linearized PQRS plasmid as competitor DNA. Upper band is the modified plasmid product and the lower  band represents cytokine mRNA product. Two bands indicates the  equivalence point of cDNA and plasmid DNA PCR products. RNA  from CD4+ cells analyzed 4 h after restimulation is shown. PQRS plasmid concentration is shown in pg/ml. For IL-2 analysis, PQRS plasmid is  shown at 100 pg/ml for induced samples, and 0.1 pg/ml for uninduced  samples to show equivalence points before normalization to hypoxanthine  phosphoribosyl transferase. (C) Kinetic analysis of mRNA synthesized by  SEA-treated and control CD4+ cells is shown. Data has been normalized  to hypoxanthine phosphoribosyl transferase.
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Figure 4: Extensively purified CD4+ cells from SEA-treated mice can synthesize cytokine mRNA. The SEA-mediated T cell anergy was reversed by FACS® sorting the CD4+ cells. (A) PCR analysis of cytokine mRNAs from SEA-treated and control CD4+ T cells after high level purification by positive selection and restimulation with 2C11 and anti-CD28 Ab. PCR products were electrophoresed on 1.5% agarose gels stained with ethidium bromide. Competitor DNA is not shown. (B) PCR products were quantitated using linearized PQRS plasmid as competitor DNA. Upper band is the modified plasmid product and the lower band represents cytokine mRNA product. Two bands indicates the equivalence point of cDNA and plasmid DNA PCR products. RNA from CD4+ cells analyzed 4 h after restimulation is shown. PQRS plasmid concentration is shown in pg/ml. For IL-2 analysis, PQRS plasmid is shown at 100 pg/ml for induced samples, and 0.1 pg/ml for uninduced samples to show equivalence points before normalization to hypoxanthine phosphoribosyl transferase. (C) Kinetic analysis of mRNA synthesized by SEA-treated and control CD4+ cells is shown. Data has been normalized to hypoxanthine phosphoribosyl transferase.

Mentions: We used sterile FACS® sorting to determine whether cytokine production was restored by purification. Sorted CD4+ cells were stimulated and analyzed for the ability to synthesize cytokine mRNA (Fig. 4). 24 h supernatants were also analyzed for cytokine content (Fig. 5).


Transferable anergy: superantigen treatment induces CD4+ T cell tolerance that is reversible and requires CD4-CD8- cells and interferon gamma.

Cauley LS, Cauley KA, Shub F, Huston G, Swain SL - J. Exp. Med. (1997)

Extensively purified CD4+ cells from SEA-treated mice can  synthesize cytokine mRNA. The SEA-mediated T cell anergy was reversed by FACS® sorting the CD4+ cells. (A) PCR analysis of cytokine  mRNAs from SEA-treated and control CD4+ T cells after high level purification by positive selection and restimulation with 2C11 and anti-CD28 Ab. PCR products were electrophoresed on 1.5% agarose gels  stained with ethidium bromide. Competitor DNA is not shown. (B)  PCR products were quantitated using linearized PQRS plasmid as competitor DNA. Upper band is the modified plasmid product and the lower  band represents cytokine mRNA product. Two bands indicates the  equivalence point of cDNA and plasmid DNA PCR products. RNA  from CD4+ cells analyzed 4 h after restimulation is shown. PQRS plasmid concentration is shown in pg/ml. For IL-2 analysis, PQRS plasmid is  shown at 100 pg/ml for induced samples, and 0.1 pg/ml for uninduced  samples to show equivalence points before normalization to hypoxanthine  phosphoribosyl transferase. (C) Kinetic analysis of mRNA synthesized by  SEA-treated and control CD4+ cells is shown. Data has been normalized  to hypoxanthine phosphoribosyl transferase.
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Figure 4: Extensively purified CD4+ cells from SEA-treated mice can synthesize cytokine mRNA. The SEA-mediated T cell anergy was reversed by FACS® sorting the CD4+ cells. (A) PCR analysis of cytokine mRNAs from SEA-treated and control CD4+ T cells after high level purification by positive selection and restimulation with 2C11 and anti-CD28 Ab. PCR products were electrophoresed on 1.5% agarose gels stained with ethidium bromide. Competitor DNA is not shown. (B) PCR products were quantitated using linearized PQRS plasmid as competitor DNA. Upper band is the modified plasmid product and the lower band represents cytokine mRNA product. Two bands indicates the equivalence point of cDNA and plasmid DNA PCR products. RNA from CD4+ cells analyzed 4 h after restimulation is shown. PQRS plasmid concentration is shown in pg/ml. For IL-2 analysis, PQRS plasmid is shown at 100 pg/ml for induced samples, and 0.1 pg/ml for uninduced samples to show equivalence points before normalization to hypoxanthine phosphoribosyl transferase. (C) Kinetic analysis of mRNA synthesized by SEA-treated and control CD4+ cells is shown. Data has been normalized to hypoxanthine phosphoribosyl transferase.
Mentions: We used sterile FACS® sorting to determine whether cytokine production was restored by purification. Sorted CD4+ cells were stimulated and analyzed for the ability to synthesize cytokine mRNA (Fig. 4). 24 h supernatants were also analyzed for cytokine content (Fig. 5).

Bottom Line: Bacterial superantigens induce peripheral unresponsiveness in CD4+ T cell populations that express appropriate Vbeta chains.However, when the SEA-treated CD4+ cells were completely purified away from all other contaminating cells, they regained the ability to proliferate and secrete cytokines.Further analysis demonstrated that interferon gamma, but not the Fas receptor, played a critical role in the suppression.

View Article: PubMed Central - PubMed

Affiliation: Trudeau Institute, Saranac Lake, New York 12983, USA.

ABSTRACT
Bacterial superantigens induce peripheral unresponsiveness in CD4+ T cell populations that express appropriate Vbeta chains. We have used Vbeta3/Valpha11 T cell receptor transgenic (Tg) mice and the Vbeta3-specific superantigen staphylococcal enterotoxin A (SEA) to further investigate the mechanisms that contribute to such unresponsiveness. As in other models, in vivo exposure to SEA rendered the Tg CD4+ cells unresponsive to subsequent restimulation in vitro with antigen or mitogens. However, when the SEA-treated CD4+ cells were completely purified away from all other contaminating cells, they regained the ability to proliferate and secrete cytokines. Moreover, enriched CD4-CD8- cells from the SEA-treated mice suppressed the responses of fresh control CD4+ cells in mixed cultures indicating that the apparent "anergy" was both transferable and reversible. Further analysis demonstrated that interferon gamma, but not the Fas receptor, played a critical role in the suppression.

Show MeSH
Related in: MedlinePlus