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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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Staining for the 2A1 antigen which is abundant in DCs in lymph node T areas. (a and b) Low and high power views for macrophages (sialoadhesin+) in the subcapsular sinus (arrows) and medulla (brown) and the 2A1 antigen in the deep cortex (T area) and to a less extent B cell follicles (blue).  (c) The deep cortex with several venules (*) is stained for the 2A1 antigen (brown) and counterstained with hematoxylin.
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Figure 2: Staining for the 2A1 antigen which is abundant in DCs in lymph node T areas. (a and b) Low and high power views for macrophages (sialoadhesin+) in the subcapsular sinus (arrows) and medulla (brown) and the 2A1 antigen in the deep cortex (T area) and to a less extent B cell follicles (blue). (c) The deep cortex with several venules (*) is stained for the 2A1 antigen (brown) and counterstained with hematoxylin.

Mentions: Fig. 2 illustrates the distribution of the 2A1 staining. This monoclonal stains granules within isolated mature DCs as described (8), but the molecule expressing the 2A1 antigen has yet to be identified. In sections, strong staining is seen on many profiles in the T cell areas, with weaker staining in the B cell areas. However, 2A1 staining is not seen on most DCs which are isolated by earlier methods from spleen and lymph node. Two other mAbs, M342 (15) and MIDC-8 (16), also do not stain freshly isolated less mature DCs but only stain after the cells have been cultured overnight.


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)

Staining for the 2A1 antigen which is abundant in DCs in lymph node T areas. (a and b) Low and high power views for macrophages (sialoadhesin+) in the subcapsular sinus (arrows) and medulla (brown) and the 2A1 antigen in the deep cortex (T area) and to a less extent B cell follicles (blue).  (c) The deep cortex with several venules (*) is stained for the 2A1 antigen (brown) and counterstained with hematoxylin.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199017&req=5

Figure 2: Staining for the 2A1 antigen which is abundant in DCs in lymph node T areas. (a and b) Low and high power views for macrophages (sialoadhesin+) in the subcapsular sinus (arrows) and medulla (brown) and the 2A1 antigen in the deep cortex (T area) and to a less extent B cell follicles (blue). (c) The deep cortex with several venules (*) is stained for the 2A1 antigen (brown) and counterstained with hematoxylin.
Mentions: Fig. 2 illustrates the distribution of the 2A1 staining. This monoclonal stains granules within isolated mature DCs as described (8), but the molecule expressing the 2A1 antigen has yet to be identified. In sections, strong staining is seen on many profiles in the T cell areas, with weaker staining in the B cell areas. However, 2A1 staining is not seen on most DCs which are isolated by earlier methods from spleen and lymph node. Two other mAbs, M342 (15) and MIDC-8 (16), also do not stain freshly isolated less mature DCs but only stain after the cells have been cultured overnight.

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