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Targeted expression of major histocompatibility complex (MHC) class II molecules demonstrates that dendritic cells can induce negative but not positive selection of thymocytes in vivo.

Brocker T, Riedinger M, Karjalainen K - J. Exp. Med. (1997)

Bottom Line: Using the CD 11c promoter we expressed MHC class II I-E molecules specifically on DC of all tissues, but not on other cell types.In contrast, it only DC expressed I-E in a class II-deficient background, positive selection of CD4+ T cells could not be observed.Thus negative, but not positive, selection events can be induced by DC in vivo.

View Article: PubMed Central - PubMed

Affiliation: Basel Institute for Immunology, Switzerland.

ABSTRACT
It is well established that lymphoid dendritic cells (DC) play an important role in the immune system. Beside their role as potent inducers of primary T cell responses, DC seem to play a crucial part as major histocompatibility complex (MHC) class II+ "interdigitating cells" in the thymus during thymocyte development. Thymic DC have been implicated in tolerance induction and also by some authors in inducing major histocompatibility complex restriction of thymocytes. Most of our knowledge about thymic DC was obtained using highly invasive and manipulatory experimental protocols such as thymus reaggregation cultures, suspension cultures, thymus grafting, and bone marrow reconstitution experiments. The DC used in those studies had to go through extensive isolation procedures or were cultured with recombinant growth factors. Since the functions of DC after these in vitro manipulations have been reported to be not identical to those of DC in vivo, we intended to establish a system that would allow us to investigate DC function avoiding artificial interferences due to handling. Here we present a transgenic mouse model in which we targeted gene expression specifically to DC. Using the CD 11c promoter we expressed MHC class II I-E molecules specifically on DC of all tissues, but not on other cell types. We report that I-E expression on thymic DC is sufficient to negatively select I-E reactive CD4+ T cells, and to a less complete extent, CD8+ T cells. In contrast, it only DC expressed I-E in a class II-deficient background, positive selection of CD4+ T cells could not be observed. Thus negative, but not positive, selection events can be induced by DC in vivo.

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Serial cryostat sections of normal adult thymus stained with  14.4.4S specific for I-E (a, c, and e) and UEA-1 (b, d, and f  ), a lectin binding specific for medullary epithelial cells (28). (a and b) Serial sections of a  wild-type B6 (I-E−) thymus. (a) I-E, no I-E staining can be detected in  the B6 background, (b) UEA-1, staining is restricted to dense aggregates  of epithelial cells in the medulla (M), while no staining can be detected in  the cortex (C). (c and d) Serial sections of B6-Eαd (I-E+) thymus, class II I-E  transgene staining is confluent on BM-derived and epithelial cells in the  medulla (M), and reticular staining in the cortex (C) is typical of epithelial  cells. UEA-1 staining (as in b) marking the medullary region (M) only.  (e and f   ) Serial sections of B6CD11c-Eαd thymus. I-E (e) transgene under  control of the CD11c promoter can be detected only in medulla (M) and  medullary-cortical junctions. No I-E staining in cortex (C) was detectable. UEA-1 ( f  ) staining is the same as for b and d. All sections were photographed at ×100.
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Figure 2: Serial cryostat sections of normal adult thymus stained with 14.4.4S specific for I-E (a, c, and e) and UEA-1 (b, d, and f  ), a lectin binding specific for medullary epithelial cells (28). (a and b) Serial sections of a wild-type B6 (I-E−) thymus. (a) I-E, no I-E staining can be detected in the B6 background, (b) UEA-1, staining is restricted to dense aggregates of epithelial cells in the medulla (M), while no staining can be detected in the cortex (C). (c and d) Serial sections of B6-Eαd (I-E+) thymus, class II I-E transgene staining is confluent on BM-derived and epithelial cells in the medulla (M), and reticular staining in the cortex (C) is typical of epithelial cells. UEA-1 staining (as in b) marking the medullary region (M) only. (e and f   ) Serial sections of B6CD11c-Eαd thymus. I-E (e) transgene under control of the CD11c promoter can be detected only in medulla (M) and medullary-cortical junctions. No I-E staining in cortex (C) was detectable. UEA-1 ( f  ) staining is the same as for b and d. All sections were photographed at ×100.

Mentions: Immunohistological analysis of thymi of the three described mouse lines is shown in Fig. 2. Serial sections were incubated with UEA-1, a fucose-specific lectin that stains medullary epithelial cells (28; Fig. 2, b, d, and f   ), and 14.4.4S to monitor transgene expression (Fig. 2, a, c, and e). While B6 thymus does not show any I-E expression (Fig. 2 a) in medulla (as localized with UEA-1 in Fig. 2 b) or cortex, the control B6-Eαd thymus (Fig. 2, c and d) could be stained with 14.4.4S (Fig. 2 c) in medulla as well as in cortical regions. While the medulla (Fig. 2 c, M) showed a strong staining due to I-E expression on its BM-derived and epithelial components (32), the cortical area (Fig. 2 c, C) stained in a typical reticular lattice pattern due to transgene expression on cortical epithelial cells (32).


Targeted expression of major histocompatibility complex (MHC) class II molecules demonstrates that dendritic cells can induce negative but not positive selection of thymocytes in vivo.

Brocker T, Riedinger M, Karjalainen K - J. Exp. Med. (1997)

Serial cryostat sections of normal adult thymus stained with  14.4.4S specific for I-E (a, c, and e) and UEA-1 (b, d, and f  ), a lectin binding specific for medullary epithelial cells (28). (a and b) Serial sections of a  wild-type B6 (I-E−) thymus. (a) I-E, no I-E staining can be detected in  the B6 background, (b) UEA-1, staining is restricted to dense aggregates  of epithelial cells in the medulla (M), while no staining can be detected in  the cortex (C). (c and d) Serial sections of B6-Eαd (I-E+) thymus, class II I-E  transgene staining is confluent on BM-derived and epithelial cells in the  medulla (M), and reticular staining in the cortex (C) is typical of epithelial  cells. UEA-1 staining (as in b) marking the medullary region (M) only.  (e and f   ) Serial sections of B6CD11c-Eαd thymus. I-E (e) transgene under  control of the CD11c promoter can be detected only in medulla (M) and  medullary-cortical junctions. No I-E staining in cortex (C) was detectable. UEA-1 ( f  ) staining is the same as for b and d. All sections were photographed at ×100.
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Figure 2: Serial cryostat sections of normal adult thymus stained with 14.4.4S specific for I-E (a, c, and e) and UEA-1 (b, d, and f  ), a lectin binding specific for medullary epithelial cells (28). (a and b) Serial sections of a wild-type B6 (I-E−) thymus. (a) I-E, no I-E staining can be detected in the B6 background, (b) UEA-1, staining is restricted to dense aggregates of epithelial cells in the medulla (M), while no staining can be detected in the cortex (C). (c and d) Serial sections of B6-Eαd (I-E+) thymus, class II I-E transgene staining is confluent on BM-derived and epithelial cells in the medulla (M), and reticular staining in the cortex (C) is typical of epithelial cells. UEA-1 staining (as in b) marking the medullary region (M) only. (e and f   ) Serial sections of B6CD11c-Eαd thymus. I-E (e) transgene under control of the CD11c promoter can be detected only in medulla (M) and medullary-cortical junctions. No I-E staining in cortex (C) was detectable. UEA-1 ( f  ) staining is the same as for b and d. All sections were photographed at ×100.
Mentions: Immunohistological analysis of thymi of the three described mouse lines is shown in Fig. 2. Serial sections were incubated with UEA-1, a fucose-specific lectin that stains medullary epithelial cells (28; Fig. 2, b, d, and f   ), and 14.4.4S to monitor transgene expression (Fig. 2, a, c, and e). While B6 thymus does not show any I-E expression (Fig. 2 a) in medulla (as localized with UEA-1 in Fig. 2 b) or cortex, the control B6-Eαd thymus (Fig. 2, c and d) could be stained with 14.4.4S (Fig. 2 c) in medulla as well as in cortical regions. While the medulla (Fig. 2 c, M) showed a strong staining due to I-E expression on its BM-derived and epithelial components (32), the cortical area (Fig. 2 c, C) stained in a typical reticular lattice pattern due to transgene expression on cortical epithelial cells (32).

Bottom Line: Using the CD 11c promoter we expressed MHC class II I-E molecules specifically on DC of all tissues, but not on other cell types.In contrast, it only DC expressed I-E in a class II-deficient background, positive selection of CD4+ T cells could not be observed.Thus negative, but not positive, selection events can be induced by DC in vivo.

View Article: PubMed Central - PubMed

Affiliation: Basel Institute for Immunology, Switzerland.

ABSTRACT
It is well established that lymphoid dendritic cells (DC) play an important role in the immune system. Beside their role as potent inducers of primary T cell responses, DC seem to play a crucial part as major histocompatibility complex (MHC) class II+ "interdigitating cells" in the thymus during thymocyte development. Thymic DC have been implicated in tolerance induction and also by some authors in inducing major histocompatibility complex restriction of thymocytes. Most of our knowledge about thymic DC was obtained using highly invasive and manipulatory experimental protocols such as thymus reaggregation cultures, suspension cultures, thymus grafting, and bone marrow reconstitution experiments. The DC used in those studies had to go through extensive isolation procedures or were cultured with recombinant growth factors. Since the functions of DC after these in vitro manipulations have been reported to be not identical to those of DC in vivo, we intended to establish a system that would allow us to investigate DC function avoiding artificial interferences due to handling. Here we present a transgenic mouse model in which we targeted gene expression specifically to DC. Using the CD 11c promoter we expressed MHC class II I-E molecules specifically on DC of all tissues, but not on other cell types. We report that I-E expression on thymic DC is sufficient to negatively select I-E reactive CD4+ T cells, and to a less complete extent, CD8+ T cells. In contrast, it only DC expressed I-E in a class II-deficient background, positive selection of CD4+ T cells could not be observed. Thus negative, but not positive, selection events can be induced by DC in vivo.

Show MeSH
Related in: MedlinePlus