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High levels of a major histocompatibility complex II-self peptide complex on dendritic cells from the T cell areas of lymph nodes.

Inaba K, Pack M, Inaba M, Sakuta H, Isdell F, Steinman RM - J. Exp. Med. (1997)

Bottom Line: Shortman. 1992.Therefore DCs within the T cell areas can be isolated.Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

View Article: PubMed Central - PubMed

Affiliation: Kyoto University, Kitashirakawa-Oiusake-cho, Kyoto 606-01, Japan.

ABSTRACT
T lymphocytes recirculate continually through the T cell areas of peripheral lymph nodes. During each passage, the T cells survey the surface of large dendritic cells (DCs), also known as interdigitating cells. However, these DCs have been difficult to release from the lymph node. By emphasizing the use of calcium-free media, as shown by Vremec et al. (Vremec, D., M. Zorbas, R. Scollay, D.J. Saunders, C.F. Ardavin, L. Wu, and K. Shortman. 1992. J. Exp. Med. 176:47-58.), we have been able to release and enrich DCs from the T cell areas. The DCs express the CD11c leukocyte integrin, the DEC-205 multilectin receptor for antigen presentation, the intracellular granule antigens which are recognized by monoclonal antibodies M342, 2A1, and MIDC-8, very high levels of MHC I and MHC II, and abundant accessory molecules such as CD40, CD54, and CD86. When examined with the Y-Ae monoclonal which recognizes complexes formed between I-Ab and a peptide derived from I-Ealpha, the T cell area DCs expressed the highest levels. The enriched DCs also stimulated a T-T hybridoma specific for this MHC II-peptide complex, and the hybridoma underwent apoptosis. Therefore DCs within the T cell areas can be isolated. Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

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Sorted lymph node DCs are potent APCs for IL-2 release  and death of Y-Ae-specific, T-T hybridomas. Function is shown for sorted  DCs and B cells from C57BL/6 and B6 × D2 F1 mice. (A) The response  was measured as IL-2 release (bottom, proliferation of conA blasts) or 3H-TdR uptake (top) by the T-T hybrid, the latter being reduced by apoptosis. ▪, C57BL/6 B cells; □, B6D2F1 B cells; --•--, C57BL/6 DCs; --○--  B6D2F1 DCs; ○, None. (B) Apoptosis was measured by identifying hypodiploid profiles using propidium iodide staining. Extensive apoptosis  occurs when the DCs are from B6D2 mice which express the Y-Ae  epitope.
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Figure 8: Sorted lymph node DCs are potent APCs for IL-2 release and death of Y-Ae-specific, T-T hybridomas. Function is shown for sorted DCs and B cells from C57BL/6 and B6 × D2 F1 mice. (A) The response was measured as IL-2 release (bottom, proliferation of conA blasts) or 3H-TdR uptake (top) by the T-T hybrid, the latter being reduced by apoptosis. ▪, C57BL/6 B cells; □, B6D2F1 B cells; --•--, C57BL/6 DCs; --○-- B6D2F1 DCs; ○, None. (B) Apoptosis was measured by identifying hypodiploid profiles using propidium iodide staining. Extensive apoptosis occurs when the DCs are from B6D2 mice which express the Y-Ae epitope.

Mentions: FACS®-purified, CD11c+ B220− DCs and CD11c− B220+ B cells were used to present the Y-Ae epitope to two T-T hybrids which recognize I-Ab/I-Eα peptide complexes (10). The DCs were >30-fold more potent in inducing IL-2 release and inducing the hybridomas to stop synthesizing DNA (Fig. 8 A), an indication of apoptosis (25). To verify that apoptosis had taken place, the cells were stained with propidium iodide and shown to have high levels of hypodiploid profiles 24 h after stimulation with DCs from the appropriate strain of mice (Fig. 8 B, compare B6D2 with B6 DCs).


High levels of a major histocompatibility complex II-self peptide complex on dendritic cells from the T cell areas of lymph nodes.

Inaba K, Pack M, Inaba M, Sakuta H, Isdell F, Steinman RM - J. Exp. Med. (1997)

Sorted lymph node DCs are potent APCs for IL-2 release  and death of Y-Ae-specific, T-T hybridomas. Function is shown for sorted  DCs and B cells from C57BL/6 and B6 × D2 F1 mice. (A) The response  was measured as IL-2 release (bottom, proliferation of conA blasts) or 3H-TdR uptake (top) by the T-T hybrid, the latter being reduced by apoptosis. ▪, C57BL/6 B cells; □, B6D2F1 B cells; --•--, C57BL/6 DCs; --○--  B6D2F1 DCs; ○, None. (B) Apoptosis was measured by identifying hypodiploid profiles using propidium iodide staining. Extensive apoptosis  occurs when the DCs are from B6D2 mice which express the Y-Ae  epitope.
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Related In: Results  -  Collection

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Figure 8: Sorted lymph node DCs are potent APCs for IL-2 release and death of Y-Ae-specific, T-T hybridomas. Function is shown for sorted DCs and B cells from C57BL/6 and B6 × D2 F1 mice. (A) The response was measured as IL-2 release (bottom, proliferation of conA blasts) or 3H-TdR uptake (top) by the T-T hybrid, the latter being reduced by apoptosis. ▪, C57BL/6 B cells; □, B6D2F1 B cells; --•--, C57BL/6 DCs; --○-- B6D2F1 DCs; ○, None. (B) Apoptosis was measured by identifying hypodiploid profiles using propidium iodide staining. Extensive apoptosis occurs when the DCs are from B6D2 mice which express the Y-Ae epitope.
Mentions: FACS®-purified, CD11c+ B220− DCs and CD11c− B220+ B cells were used to present the Y-Ae epitope to two T-T hybrids which recognize I-Ab/I-Eα peptide complexes (10). The DCs were >30-fold more potent in inducing IL-2 release and inducing the hybridomas to stop synthesizing DNA (Fig. 8 A), an indication of apoptosis (25). To verify that apoptosis had taken place, the cells were stained with propidium iodide and shown to have high levels of hypodiploid profiles 24 h after stimulation with DCs from the appropriate strain of mice (Fig. 8 B, compare B6D2 with B6 DCs).

Bottom Line: Shortman. 1992.Therefore DCs within the T cell areas can be isolated.Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

View Article: PubMed Central - PubMed

Affiliation: Kyoto University, Kitashirakawa-Oiusake-cho, Kyoto 606-01, Japan.

ABSTRACT
T lymphocytes recirculate continually through the T cell areas of peripheral lymph nodes. During each passage, the T cells survey the surface of large dendritic cells (DCs), also known as interdigitating cells. However, these DCs have been difficult to release from the lymph node. By emphasizing the use of calcium-free media, as shown by Vremec et al. (Vremec, D., M. Zorbas, R. Scollay, D.J. Saunders, C.F. Ardavin, L. Wu, and K. Shortman. 1992. J. Exp. Med. 176:47-58.), we have been able to release and enrich DCs from the T cell areas. The DCs express the CD11c leukocyte integrin, the DEC-205 multilectin receptor for antigen presentation, the intracellular granule antigens which are recognized by monoclonal antibodies M342, 2A1, and MIDC-8, very high levels of MHC I and MHC II, and abundant accessory molecules such as CD40, CD54, and CD86. When examined with the Y-Ae monoclonal which recognizes complexes formed between I-Ab and a peptide derived from I-Ealpha, the T cell area DCs expressed the highest levels. The enriched DCs also stimulated a T-T hybridoma specific for this MHC II-peptide complex, and the hybridoma underwent apoptosis. Therefore DCs within the T cell areas can be isolated. Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

Show MeSH
Related in: MedlinePlus