Limits...
Immune tolerance after delivery of dying cells to dendritic cells in situ.

Liu K, Iyoda T, Saternus M, Kimura Y, Inaba K, Steinman RM - J. Exp. Med. (2002)

Bottom Line: After ingestion and presentation of cell-associated OVA by the CD8(+) subset of dendritic cells in situ, large numbers of antigen-reactive, CD8(+) T cell receptor (TCR) transgenic T lymphocytes were driven into cell cycle, but then the T cells were deleted.In contrast to observations made in the steady state, the antigen-reactive T cells expanded in numbers for 1-2 wk and produced large amounts of interleukin 2 and interferon gamma, while the animals retained responsiveness to antigen rechallenge.The specific tolerance that develops when dendritic cells process self tissues in the steady state should prevent or reduce the development of autoimmunity when dying cells are subsequently processed during infection.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, Chris Browne Center for Immunology and Immune Diseases, The Rockefeller University, New York, NY 10021, USA.

ABSTRACT
Peripheral immune tolerance is believed to be induced by the processing and presentation of self-tissues that die during physiologic tissue turnover. To examine the mechanism that mediates tolerance, we injected mice with dying syngeneic TAP(-/-) splenocytes loaded with small amounts of the protein antigen, ovalbumin (OVA). After ingestion and presentation of cell-associated OVA by the CD8(+) subset of dendritic cells in situ, large numbers of antigen-reactive, CD8(+) T cell receptor (TCR) transgenic T lymphocytes were driven into cell cycle, but then the T cells were deleted. The animals were also tolerant to challenge with OVA in complete Freund's adjuvant. An agonistic anti-CD40 monoclonal antibody was then administered together with the OVA-loaded splenocytes, so that the dendritic cells in the recipient mice would mature. In contrast to observations made in the steady state, the antigen-reactive T cells expanded in numbers for 1-2 wk and produced large amounts of interleukin 2 and interferon gamma, while the animals retained responsiveness to antigen rechallenge. The specific tolerance that develops when dendritic cells process self tissues in the steady state should prevent or reduce the development of autoimmunity when dying cells are subsequently processed during infection.

Show MeSH

Related in: MedlinePlus

Tolerance of mice given OVA-loaded TAP−/− splenocytes to OVA challenge with CFA. (A) 2 d after injection of 0.3 × 106 OT-I T cells, B6 mice were injected with 25 × 106 OVA-loaded or unloaded TAP−/− splenocytes intravenously. Some mice were given OVA-loaded TAP−/− splenocytes once (day –6) and others twice (days –6 and 0). 9 d after the last injection of OVA-loaded TAP−/− splenocytes, the mice were challenged with OVA in CFA, and total numbers of CD8+ CD45.1+ OT-I cells in draining (brachial) and distal (inguinal) nodes were measured by FACS® (left) as in Fig. 3. Enriched CD8+ T cells using magnetic microbeads® also were cocultured with the graded doses (inset) of SIINFEKL-pulsed spleen cells after γ-irradiation at 15 Gy. T cell proliferation was measured by 3H-TdR uptake at 38–48 h (middle), and IFN-γ–secreting cells by ELISPOT at 36 h (right). (B) Immunity develops if OVA-loaded splenocytes are presented along with anti-CD40 agonistic antibody, assessed as in B with ELISPOT (right panel) and T cell proliferation (38–48 h; left panel). The data in A and B are representative of three experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2194037&req=5

fig4: Tolerance of mice given OVA-loaded TAP−/− splenocytes to OVA challenge with CFA. (A) 2 d after injection of 0.3 × 106 OT-I T cells, B6 mice were injected with 25 × 106 OVA-loaded or unloaded TAP−/− splenocytes intravenously. Some mice were given OVA-loaded TAP−/− splenocytes once (day –6) and others twice (days –6 and 0). 9 d after the last injection of OVA-loaded TAP−/− splenocytes, the mice were challenged with OVA in CFA, and total numbers of CD8+ CD45.1+ OT-I cells in draining (brachial) and distal (inguinal) nodes were measured by FACS® (left) as in Fig. 3. Enriched CD8+ T cells using magnetic microbeads® also were cocultured with the graded doses (inset) of SIINFEKL-pulsed spleen cells after γ-irradiation at 15 Gy. T cell proliferation was measured by 3H-TdR uptake at 38–48 h (middle), and IFN-γ–secreting cells by ELISPOT at 36 h (right). (B) Immunity develops if OVA-loaded splenocytes are presented along with anti-CD40 agonistic antibody, assessed as in B with ELISPOT (right panel) and T cell proliferation (38–48 h; left panel). The data in A and B are representative of three experiments.

Mentions: To determine if systemic tolerance had been induced by DCs presenting cell-associated antigen in the steady-state, we rechallenged the mice with OVA in CFA. Mice were injected with OVA-charged splenocytes either once (day –6) or twice (day –6 and 0), and 9 d later challenged with OVA in CFA. In mice given TAP−/− splenocytes without OVA protein, a strong immune response was evident 3 d after the challenge with OVA in CFA, by three criteria (Fig. 4 A, top row). These were (a) increased OT-I T cell numbers in the lymph nodes draining the site of OVA/CFA injection, (b) strong proliferation to OVA challenge in culture, and (c) the appearance of many IFN-γ–secreting cells in ELISPOT assays. In contrast, mice exposed to one or two doses of OVA-bearing splenocytes became unresponsive to challenge with CFA/OVA (Fig. 4 A). This tolerance was antigen specific, as coadministered F5 TCR transgenic cells specific for a viral peptide (see Fig. 1 E) were not deleted (data not depicted).


Immune tolerance after delivery of dying cells to dendritic cells in situ.

Liu K, Iyoda T, Saternus M, Kimura Y, Inaba K, Steinman RM - J. Exp. Med. (2002)

Tolerance of mice given OVA-loaded TAP−/− splenocytes to OVA challenge with CFA. (A) 2 d after injection of 0.3 × 106 OT-I T cells, B6 mice were injected with 25 × 106 OVA-loaded or unloaded TAP−/− splenocytes intravenously. Some mice were given OVA-loaded TAP−/− splenocytes once (day –6) and others twice (days –6 and 0). 9 d after the last injection of OVA-loaded TAP−/− splenocytes, the mice were challenged with OVA in CFA, and total numbers of CD8+ CD45.1+ OT-I cells in draining (brachial) and distal (inguinal) nodes were measured by FACS® (left) as in Fig. 3. Enriched CD8+ T cells using magnetic microbeads® also were cocultured with the graded doses (inset) of SIINFEKL-pulsed spleen cells after γ-irradiation at 15 Gy. T cell proliferation was measured by 3H-TdR uptake at 38–48 h (middle), and IFN-γ–secreting cells by ELISPOT at 36 h (right). (B) Immunity develops if OVA-loaded splenocytes are presented along with anti-CD40 agonistic antibody, assessed as in B with ELISPOT (right panel) and T cell proliferation (38–48 h; left panel). The data in A and B are representative of three experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194037&req=5

fig4: Tolerance of mice given OVA-loaded TAP−/− splenocytes to OVA challenge with CFA. (A) 2 d after injection of 0.3 × 106 OT-I T cells, B6 mice were injected with 25 × 106 OVA-loaded or unloaded TAP−/− splenocytes intravenously. Some mice were given OVA-loaded TAP−/− splenocytes once (day –6) and others twice (days –6 and 0). 9 d after the last injection of OVA-loaded TAP−/− splenocytes, the mice were challenged with OVA in CFA, and total numbers of CD8+ CD45.1+ OT-I cells in draining (brachial) and distal (inguinal) nodes were measured by FACS® (left) as in Fig. 3. Enriched CD8+ T cells using magnetic microbeads® also were cocultured with the graded doses (inset) of SIINFEKL-pulsed spleen cells after γ-irradiation at 15 Gy. T cell proliferation was measured by 3H-TdR uptake at 38–48 h (middle), and IFN-γ–secreting cells by ELISPOT at 36 h (right). (B) Immunity develops if OVA-loaded splenocytes are presented along with anti-CD40 agonistic antibody, assessed as in B with ELISPOT (right panel) and T cell proliferation (38–48 h; left panel). The data in A and B are representative of three experiments.
Mentions: To determine if systemic tolerance had been induced by DCs presenting cell-associated antigen in the steady-state, we rechallenged the mice with OVA in CFA. Mice were injected with OVA-charged splenocytes either once (day –6) or twice (day –6 and 0), and 9 d later challenged with OVA in CFA. In mice given TAP−/− splenocytes without OVA protein, a strong immune response was evident 3 d after the challenge with OVA in CFA, by three criteria (Fig. 4 A, top row). These were (a) increased OT-I T cell numbers in the lymph nodes draining the site of OVA/CFA injection, (b) strong proliferation to OVA challenge in culture, and (c) the appearance of many IFN-γ–secreting cells in ELISPOT assays. In contrast, mice exposed to one or two doses of OVA-bearing splenocytes became unresponsive to challenge with CFA/OVA (Fig. 4 A). This tolerance was antigen specific, as coadministered F5 TCR transgenic cells specific for a viral peptide (see Fig. 1 E) were not deleted (data not depicted).

Bottom Line: After ingestion and presentation of cell-associated OVA by the CD8(+) subset of dendritic cells in situ, large numbers of antigen-reactive, CD8(+) T cell receptor (TCR) transgenic T lymphocytes were driven into cell cycle, but then the T cells were deleted.In contrast to observations made in the steady state, the antigen-reactive T cells expanded in numbers for 1-2 wk and produced large amounts of interleukin 2 and interferon gamma, while the animals retained responsiveness to antigen rechallenge.The specific tolerance that develops when dendritic cells process self tissues in the steady state should prevent or reduce the development of autoimmunity when dying cells are subsequently processed during infection.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, Chris Browne Center for Immunology and Immune Diseases, The Rockefeller University, New York, NY 10021, USA.

ABSTRACT
Peripheral immune tolerance is believed to be induced by the processing and presentation of self-tissues that die during physiologic tissue turnover. To examine the mechanism that mediates tolerance, we injected mice with dying syngeneic TAP(-/-) splenocytes loaded with small amounts of the protein antigen, ovalbumin (OVA). After ingestion and presentation of cell-associated OVA by the CD8(+) subset of dendritic cells in situ, large numbers of antigen-reactive, CD8(+) T cell receptor (TCR) transgenic T lymphocytes were driven into cell cycle, but then the T cells were deleted. The animals were also tolerant to challenge with OVA in complete Freund's adjuvant. An agonistic anti-CD40 monoclonal antibody was then administered together with the OVA-loaded splenocytes, so that the dendritic cells in the recipient mice would mature. In contrast to observations made in the steady state, the antigen-reactive T cells expanded in numbers for 1-2 wk and produced large amounts of interleukin 2 and interferon gamma, while the animals retained responsiveness to antigen rechallenge. The specific tolerance that develops when dendritic cells process self tissues in the steady state should prevent or reduce the development of autoimmunity when dying cells are subsequently processed during infection.

Show MeSH
Related in: MedlinePlus