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In vivo-activated CD4 T cells upregulate CXC chemokine receptor 5 and reprogram their response to lymphoid chemokines.

Ansel KM, McHeyzer-Williams LJ, Ngo VN, McHeyzer-Williams MG, Cyster JG - J. Exp. Med. (1999)

Bottom Line: At the same time, the CXCR5(hi) cells showed reduced responsiveness to the T zone chemokines, Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC) and secondary lymphoid tissue chemokine (SLC).After adoptive transfer, CXCR5(hi) CD4 T cells did not migrate to follicles, indicating that additional changes may occur after immunization that help direct T cells to follicles.Taken together, our findings indicate that reprogramming of responsiveness to constitutively expressed lymphoid tissue chemokines plays an important role in T cell migration to the B cell compartment of lymphoid tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA.

ABSTRACT
Migration of antigen-activated CD4 T cells to B cell areas of lymphoid tissues is important for mounting T cell-dependent antibody responses. Here we show that CXC chemokine receptor (CXCR)5, the receptor for B lymphocyte chemoattractant (BLC), is upregulated on antigen-specific CD4 T cells in vivo when animals are immunized under conditions that promote T cell migration to follicles. In situ hybridization of secondary follicles for BLC showed high expression in mantle zones and low expression in germinal centers. When tested directly ex vivo, CXCR5(hi) T cells exhibited a vigorous chemotactic response to BLC. At the same time, the CXCR5(hi) cells showed reduced responsiveness to the T zone chemokines, Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC) and secondary lymphoid tissue chemokine (SLC). After adoptive transfer, CXCR5(hi) CD4 T cells did not migrate to follicles, indicating that additional changes may occur after immunization that help direct T cells to follicles. To further explore whether T cells could acquire an intrinsic ability to migrate to follicles, CD4(-)CD8(-) double negative (DN) T cells from MRL-lpr mice were studied. These T cells normally accumulate within follicles of MRL-lpr mice. Upon transfer to wild-type recipients, DN T cells migrated to follicle proximal regions in all secondary lymphoid tissues. Taken together, our findings indicate that reprogramming of responsiveness to constitutively expressed lymphoid tissue chemokines plays an important role in T cell migration to the B cell compartment of lymphoid tissues.

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CXCR5 expression and chemotactic response profile of DN T cells from MRL-lpr mice. (A) CXCR5 expression on DN (Thy1+B220+) and conventional (Thy1+B220−) T cells. DN T cells stained with the secondary antibody alone (no 1° Ab) are shown as a control. (B and C) Chemotaxis of a 3:1 mixture of MRL-lpr and B6 splenocytes in response to (B) BLC and (C) a panel of lymphoid chemokines. Results are expressed as percentage of input cells transmigrated for DN T cells (•) and conventional T cells (○). Chemokine concentrations in C: BLC, ELC, and SLC, 1 μg/ml; SDF1, 0.3 μg/ml. MRL-lpr mice were old 5 mo at the time of analysis. Lines (B) and bars (C) represent means of duplicate transwells. Results in A are representative of three, and in B and C of two independent experiments.
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Figure 7: CXCR5 expression and chemotactic response profile of DN T cells from MRL-lpr mice. (A) CXCR5 expression on DN (Thy1+B220+) and conventional (Thy1+B220−) T cells. DN T cells stained with the secondary antibody alone (no 1° Ab) are shown as a control. (B and C) Chemotaxis of a 3:1 mixture of MRL-lpr and B6 splenocytes in response to (B) BLC and (C) a panel of lymphoid chemokines. Results are expressed as percentage of input cells transmigrated for DN T cells (•) and conventional T cells (○). Chemokine concentrations in C: BLC, ELC, and SLC, 1 μg/ml; SDF1, 0.3 μg/ml. MRL-lpr mice were old 5 mo at the time of analysis. Lines (B) and bars (C) represent means of duplicate transwells. Results in A are representative of three, and in B and C of two independent experiments.

Mentions: The studies above demonstrate a tight relationship between CD4 T cell upregulation of CXCR5 expression, acquisition of BLC responsiveness, and migration into lymphoid follicles during an immune response. However, they do not establish whether intrinsic changes in the T cell are sufficient to direct these cells to follicles or whether additional (extrinsic) changes that accompany the adjuvant-induced immune response are also needed. Although many of the CD4 T cells in aged mice express CXCR5 (see Fig. 4 C), immunohistochemical analysis did not reveal a significantly greater number of T cells in follicles in aged mice compared with young mice (data not shown). When CXCR5hi CD4 cells were transferred from aged to young mice, they were found to localize within the T zone, with only occasional cells migrating into follicles (Fig. 6 A). These findings indicate that expression of CXCR5 is not sufficient to direct all types of T cells into B cell follicles. However, in contrast to aged normal mice, aged MRL-lpr mice contain very large numbers of T cells in a follicular distribution 3940. These CD3+ T cells are unusual in lacking CD4 and CD8 and in expressing B220 3940. Flow cytometric analysis showed that they also express high surface CXCR5 (Fig. 7 A), and in in vitro chemotaxis assays they demonstrated a robust response to BLC (Fig. 7 B) and a reduced response to ELC and SLC (Fig. 7 C). Importantly, the BLC dose–response curve of the DN T cells was typical of CXCR5+ CD4 T cells (compare Fig. 7 B and Fig. 4A and Fig. D) and not of B cells (see Fig. 4 D). Although the follicular location of the DN T cells in MRL-lpr mice suggests these cells have acquired the intrinsic ability to migrate to follicles, MRL-lpr mice have multiple immunological abnormalities, and it was possible that homing of DN T cells to follicles was dependent on extrinsic changes in the lymphoid tissues. To test this directly, DN T cells were purified from LNs of MRL-lpr mice, labeled with CFSE, and transferred to normal syngeneic MRL mice. Strikingly, the transferred T cells migrated to regions proximal to B cell areas in all secondary lymphoid tissues of the recipient mice (Fig. 6). Differences in the distribution of the cells were noted in the different tissues. In the spleen, the T cells homed to the outer rim of the follicles, especially near the marginal zone bridging channels, and often the cells appeared in contact with marginal metallophilic macrophages (Fig. 6 B). In LNs the T cells homed to perifollicular and interfollicular locations (Fig. 6 C), and in Peyer's patches the cells were seen to circle the whole follicular area (Fig. 6 D). These observations demonstrate that T cells can acquire the intrinsic ability to migrate to the boundaries of lymphoid follicles.


In vivo-activated CD4 T cells upregulate CXC chemokine receptor 5 and reprogram their response to lymphoid chemokines.

Ansel KM, McHeyzer-Williams LJ, Ngo VN, McHeyzer-Williams MG, Cyster JG - J. Exp. Med. (1999)

CXCR5 expression and chemotactic response profile of DN T cells from MRL-lpr mice. (A) CXCR5 expression on DN (Thy1+B220+) and conventional (Thy1+B220−) T cells. DN T cells stained with the secondary antibody alone (no 1° Ab) are shown as a control. (B and C) Chemotaxis of a 3:1 mixture of MRL-lpr and B6 splenocytes in response to (B) BLC and (C) a panel of lymphoid chemokines. Results are expressed as percentage of input cells transmigrated for DN T cells (•) and conventional T cells (○). Chemokine concentrations in C: BLC, ELC, and SLC, 1 μg/ml; SDF1, 0.3 μg/ml. MRL-lpr mice were old 5 mo at the time of analysis. Lines (B) and bars (C) represent means of duplicate transwells. Results in A are representative of three, and in B and C of two independent experiments.
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Related In: Results  -  Collection

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Figure 7: CXCR5 expression and chemotactic response profile of DN T cells from MRL-lpr mice. (A) CXCR5 expression on DN (Thy1+B220+) and conventional (Thy1+B220−) T cells. DN T cells stained with the secondary antibody alone (no 1° Ab) are shown as a control. (B and C) Chemotaxis of a 3:1 mixture of MRL-lpr and B6 splenocytes in response to (B) BLC and (C) a panel of lymphoid chemokines. Results are expressed as percentage of input cells transmigrated for DN T cells (•) and conventional T cells (○). Chemokine concentrations in C: BLC, ELC, and SLC, 1 μg/ml; SDF1, 0.3 μg/ml. MRL-lpr mice were old 5 mo at the time of analysis. Lines (B) and bars (C) represent means of duplicate transwells. Results in A are representative of three, and in B and C of two independent experiments.
Mentions: The studies above demonstrate a tight relationship between CD4 T cell upregulation of CXCR5 expression, acquisition of BLC responsiveness, and migration into lymphoid follicles during an immune response. However, they do not establish whether intrinsic changes in the T cell are sufficient to direct these cells to follicles or whether additional (extrinsic) changes that accompany the adjuvant-induced immune response are also needed. Although many of the CD4 T cells in aged mice express CXCR5 (see Fig. 4 C), immunohistochemical analysis did not reveal a significantly greater number of T cells in follicles in aged mice compared with young mice (data not shown). When CXCR5hi CD4 cells were transferred from aged to young mice, they were found to localize within the T zone, with only occasional cells migrating into follicles (Fig. 6 A). These findings indicate that expression of CXCR5 is not sufficient to direct all types of T cells into B cell follicles. However, in contrast to aged normal mice, aged MRL-lpr mice contain very large numbers of T cells in a follicular distribution 3940. These CD3+ T cells are unusual in lacking CD4 and CD8 and in expressing B220 3940. Flow cytometric analysis showed that they also express high surface CXCR5 (Fig. 7 A), and in in vitro chemotaxis assays they demonstrated a robust response to BLC (Fig. 7 B) and a reduced response to ELC and SLC (Fig. 7 C). Importantly, the BLC dose–response curve of the DN T cells was typical of CXCR5+ CD4 T cells (compare Fig. 7 B and Fig. 4A and Fig. D) and not of B cells (see Fig. 4 D). Although the follicular location of the DN T cells in MRL-lpr mice suggests these cells have acquired the intrinsic ability to migrate to follicles, MRL-lpr mice have multiple immunological abnormalities, and it was possible that homing of DN T cells to follicles was dependent on extrinsic changes in the lymphoid tissues. To test this directly, DN T cells were purified from LNs of MRL-lpr mice, labeled with CFSE, and transferred to normal syngeneic MRL mice. Strikingly, the transferred T cells migrated to regions proximal to B cell areas in all secondary lymphoid tissues of the recipient mice (Fig. 6). Differences in the distribution of the cells were noted in the different tissues. In the spleen, the T cells homed to the outer rim of the follicles, especially near the marginal zone bridging channels, and often the cells appeared in contact with marginal metallophilic macrophages (Fig. 6 B). In LNs the T cells homed to perifollicular and interfollicular locations (Fig. 6 C), and in Peyer's patches the cells were seen to circle the whole follicular area (Fig. 6 D). These observations demonstrate that T cells can acquire the intrinsic ability to migrate to the boundaries of lymphoid follicles.

Bottom Line: At the same time, the CXCR5(hi) cells showed reduced responsiveness to the T zone chemokines, Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC) and secondary lymphoid tissue chemokine (SLC).After adoptive transfer, CXCR5(hi) CD4 T cells did not migrate to follicles, indicating that additional changes may occur after immunization that help direct T cells to follicles.Taken together, our findings indicate that reprogramming of responsiveness to constitutively expressed lymphoid tissue chemokines plays an important role in T cell migration to the B cell compartment of lymphoid tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA.

ABSTRACT
Migration of antigen-activated CD4 T cells to B cell areas of lymphoid tissues is important for mounting T cell-dependent antibody responses. Here we show that CXC chemokine receptor (CXCR)5, the receptor for B lymphocyte chemoattractant (BLC), is upregulated on antigen-specific CD4 T cells in vivo when animals are immunized under conditions that promote T cell migration to follicles. In situ hybridization of secondary follicles for BLC showed high expression in mantle zones and low expression in germinal centers. When tested directly ex vivo, CXCR5(hi) T cells exhibited a vigorous chemotactic response to BLC. At the same time, the CXCR5(hi) cells showed reduced responsiveness to the T zone chemokines, Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC) and secondary lymphoid tissue chemokine (SLC). After adoptive transfer, CXCR5(hi) CD4 T cells did not migrate to follicles, indicating that additional changes may occur after immunization that help direct T cells to follicles. To further explore whether T cells could acquire an intrinsic ability to migrate to follicles, CD4(-)CD8(-) double negative (DN) T cells from MRL-lpr mice were studied. These T cells normally accumulate within follicles of MRL-lpr mice. Upon transfer to wild-type recipients, DN T cells migrated to follicle proximal regions in all secondary lymphoid tissues. Taken together, our findings indicate that reprogramming of responsiveness to constitutively expressed lymphoid tissue chemokines plays an important role in T cell migration to the B cell compartment of lymphoid tissues.

Show MeSH