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Mice lacking expression of secondary lymphoid organ chemokine have defects in lymphocyte homing and dendritic cell localization.

Gunn MD, Kyuwa S, Tam C, Kakiuchi T, Matsuzawa A, Williams LT, Nakano H - J. Exp. Med. (1999)

Bottom Line: Secondary lymphoid organ chemokine (SLC) is expressed in high endothelial venules and in T cell zones of spleen and lymph nodes (LNs) and strongly attracts naive T cells.DC migration to LNs after contact sensitization is also substantially reduced.The plt mutation maps to the SLC locus; however, the sequence of SLC introns and exons in plt mice is normal.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Institute, University of California San Francisco, San Francisco, California 94143, USA. michael.gunn@duke.edu

ABSTRACT
Secondary lymphoid organ chemokine (SLC) is expressed in high endothelial venules and in T cell zones of spleen and lymph nodes (LNs) and strongly attracts naive T cells. In mice homozygous for the paucity of lymph node T cell (plt) mutation, naive T cells fail to home to LNs or the lymphoid regions of spleen. Here we demonstrate that expression of SLC is undetectable in plt mice. In addition to the defect in T cell homing, we demonstrate that dendritic cells (DCs) fail to accumulate in spleen and LN T cell zones of plt mice. DC migration to LNs after contact sensitization is also substantially reduced. The physiologic significance of these abnormalities in plt mice is indicated by a markedly increased sensitivity to infection with murine hepatitis virus. The plt mutation maps to the SLC locus; however, the sequence of SLC introns and exons in plt mice is normal. These findings suggest that the abnormalities in plt mice are due to a genetic defect in the expression of SLC and that SLC mediates the entry of naive T cells and antigen-stimulated DCs into the T cell zones of secondary lymphoid organs.

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Decreased migration of skin DCs to LNs in plt mice after  contact sensitization with FITC. (A) The shaved abdomens of +/+ and  plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and  analyzed by flow cytometry. A decreased number of large FITC+ cells  (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS® profile showing a marked decrease of CD11c+ FITC+ cells in plt mice after FITC skin painting. (C)  Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by  SA-PerCP, and analyzed by flow cytometry. Only large FITC+ cells  (boxed areas in A) are shown. (D) The number of FITC+ DCs (boxed areas  in A) that accumulate in LNs after skin painting with 2 mg FITC is  reduced in plt mice. Numbers represent mean ± SD (n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal  LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A+ B220− cells.
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Figure 4: Decreased migration of skin DCs to LNs in plt mice after contact sensitization with FITC. (A) The shaved abdomens of +/+ and plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and analyzed by flow cytometry. A decreased number of large FITC+ cells (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS® profile showing a marked decrease of CD11c+ FITC+ cells in plt mice after FITC skin painting. (C) Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by SA-PerCP, and analyzed by flow cytometry. Only large FITC+ cells (boxed areas in A) are shown. (D) The number of FITC+ DCs (boxed areas in A) that accumulate in LNs after skin painting with 2 mg FITC is reduced in plt mice. Numbers represent mean ± SD (n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A+ B220− cells.

Mentions: Interdigitating DCs in the T cell zones of LNs arise from DCs in the periphery which migrate to this area via the lymph after activation. To determine if the paucity of interdigitating cells observed in plt mice was due to a defect in the migration of DCs to LNs, we examined this migration after contact sensitization. 24 h after skin painting with 2 mg FITC, the frequency of FITC+ cells in the draining LNs of plt mice was markedly reduced compared with +/+ mice (Fig. 4 A). The identity of these cells as DCs was confirmed by their characteristic forward and side scatter profiles, their staining with anti-CD11c (Fig. 4 B) and anti–I-A (Fig. 4 C), and their low buoyant density. A comparison of eight FITC-painted plt mice with controls at 24 h revealed a 75% decrease in the number of FITC+ DCs in draining LNs (Fig. 4 D).


Mice lacking expression of secondary lymphoid organ chemokine have defects in lymphocyte homing and dendritic cell localization.

Gunn MD, Kyuwa S, Tam C, Kakiuchi T, Matsuzawa A, Williams LT, Nakano H - J. Exp. Med. (1999)

Decreased migration of skin DCs to LNs in plt mice after  contact sensitization with FITC. (A) The shaved abdomens of +/+ and  plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and  analyzed by flow cytometry. A decreased number of large FITC+ cells  (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS® profile showing a marked decrease of CD11c+ FITC+ cells in plt mice after FITC skin painting. (C)  Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by  SA-PerCP, and analyzed by flow cytometry. Only large FITC+ cells  (boxed areas in A) are shown. (D) The number of FITC+ DCs (boxed areas  in A) that accumulate in LNs after skin painting with 2 mg FITC is  reduced in plt mice. Numbers represent mean ± SD (n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal  LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A+ B220− cells.
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Related In: Results  -  Collection

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Figure 4: Decreased migration of skin DCs to LNs in plt mice after contact sensitization with FITC. (A) The shaved abdomens of +/+ and plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and analyzed by flow cytometry. A decreased number of large FITC+ cells (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS® profile showing a marked decrease of CD11c+ FITC+ cells in plt mice after FITC skin painting. (C) Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by SA-PerCP, and analyzed by flow cytometry. Only large FITC+ cells (boxed areas in A) are shown. (D) The number of FITC+ DCs (boxed areas in A) that accumulate in LNs after skin painting with 2 mg FITC is reduced in plt mice. Numbers represent mean ± SD (n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A+ B220− cells.
Mentions: Interdigitating DCs in the T cell zones of LNs arise from DCs in the periphery which migrate to this area via the lymph after activation. To determine if the paucity of interdigitating cells observed in plt mice was due to a defect in the migration of DCs to LNs, we examined this migration after contact sensitization. 24 h after skin painting with 2 mg FITC, the frequency of FITC+ cells in the draining LNs of plt mice was markedly reduced compared with +/+ mice (Fig. 4 A). The identity of these cells as DCs was confirmed by their characteristic forward and side scatter profiles, their staining with anti-CD11c (Fig. 4 B) and anti–I-A (Fig. 4 C), and their low buoyant density. A comparison of eight FITC-painted plt mice with controls at 24 h revealed a 75% decrease in the number of FITC+ DCs in draining LNs (Fig. 4 D).

Bottom Line: Secondary lymphoid organ chemokine (SLC) is expressed in high endothelial venules and in T cell zones of spleen and lymph nodes (LNs) and strongly attracts naive T cells.DC migration to LNs after contact sensitization is also substantially reduced.The plt mutation maps to the SLC locus; however, the sequence of SLC introns and exons in plt mice is normal.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Institute, University of California San Francisco, San Francisco, California 94143, USA. michael.gunn@duke.edu

ABSTRACT
Secondary lymphoid organ chemokine (SLC) is expressed in high endothelial venules and in T cell zones of spleen and lymph nodes (LNs) and strongly attracts naive T cells. In mice homozygous for the paucity of lymph node T cell (plt) mutation, naive T cells fail to home to LNs or the lymphoid regions of spleen. Here we demonstrate that expression of SLC is undetectable in plt mice. In addition to the defect in T cell homing, we demonstrate that dendritic cells (DCs) fail to accumulate in spleen and LN T cell zones of plt mice. DC migration to LNs after contact sensitization is also substantially reduced. The physiologic significance of these abnormalities in plt mice is indicated by a markedly increased sensitivity to infection with murine hepatitis virus. The plt mutation maps to the SLC locus; however, the sequence of SLC introns and exons in plt mice is normal. These findings suggest that the abnormalities in plt mice are due to a genetic defect in the expression of SLC and that SLC mediates the entry of naive T cells and antigen-stimulated DCs into the T cell zones of secondary lymphoid organs.

Show MeSH
Related in: MedlinePlus