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Molecular basis for hematopoietic/mesenchymal interaction during initiation of Peyer's patch organogenesis.

Honda K, Nakano H, Yoshida H, Nishikawa S, Rennert P, Ikuta K, Tamechika M, Yamaguchi K, Fukumoto T, Chiba T, Nishikawa SI - J. Exp. Med. (2001)

Bottom Line: The lymphoid population expresses CXCR5 and has a strong chemotactic response to B lymphocyte chemoattractant (BLC).Importantly, chemokines produced by VCAM-1(+)ICAM-1(+) nonlymphoid cells mediate the recruitment of lymphoid cells.Thus, promotion of PP development relies on mutual interaction between mesenchymal and lymphoid cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Syogoin-Kawaharacho 53, Sakyo-ku, Kyoto 606-8507, Japan. khonda@virus.kyoto-u.ac.jp

ABSTRACT
Mice deficient in lymphotoxin beta receptor (LTbetaR) or interleukin 7 receptor alpha (IL-7Ralpha) lack Peyer's patches (PPs). Deficiency in CXC chemokine receptor 5 (CXCR5) also severely affects the development of PPs. A molecular network involving these three signaling pathways has been implicated in PP organogenesis, but it remains unclear how they are connected during this process. We have shown that PP organogenesis is initiated at sites containing IL-7Ralpha(+) lymphoid cells and vascular cell adhesion molecule (VCAM)-1/intercellular adhesion molecule (ICAM)-1 expressing nonlymphoid elements. Here we characterize these lymphoid and nonlymphoid components in terms of chemokine signals. The lymphoid population expresses CXCR5 and has a strong chemotactic response to B lymphocyte chemoattractant (BLC). Importantly, chemokines produced by VCAM-1(+)ICAM-1(+) nonlymphoid cells mediate the recruitment of lymphoid cells. Furthermore, we show that these VCAM-1(+)ICAM-1(+) cells are mesenchymal cells that are activated by lymphoid cells through the LTbetaR to express adhesion molecules and chemokines. Thus, promotion of PP development relies on mutual interaction between mesenchymal and lymphoid cells.

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Chemotactic activity of BLC, ELC, and SLC on CD4+CD3−IL-7Rα1 cells. (A) Whole-mount immunostaining analysis of E15.5, E16.5, and E17.5 gut with anti-CD4 mAb. (B) RT-PCR analysis on CXCR5 and CCR7 in FACS®-sorted CD4+CD3−IL-7Rα1 cells from intestine at E17.5. The numbers of cells subjected to RT-PCR are 1,000, 200, 40, and 8 cells from left to right in each panel. Specific primers for CD4, IL-7Rα, and CD3-ε were used as positive and negative controls. (C) CD4+ cells were prepared with Dynabeads and placed in the chemotaxis chamber in duplicates. ELC and SLC maximally attracted CD4+ cells at 100–300 ng/ml. Results are expressed as the percentage of input cells migrating to the lower chamber containing BLC (1 μg/ml), ELC (300 ng/ml), or SLC (300 ng/ml). Data from individual wells are shown as filled diamonds and means as bars. The results are representative of three independent experiments. (D) Chemotactic response to BLC by CD4+ cells from intestine at E17.5 (filled circles) or B cells from Spl (open squares). Lines represent the averages of duplicated Transwells. The results are representative of at least three independent experiments.
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Figure 1: Chemotactic activity of BLC, ELC, and SLC on CD4+CD3−IL-7Rα1 cells. (A) Whole-mount immunostaining analysis of E15.5, E16.5, and E17.5 gut with anti-CD4 mAb. (B) RT-PCR analysis on CXCR5 and CCR7 in FACS®-sorted CD4+CD3−IL-7Rα1 cells from intestine at E17.5. The numbers of cells subjected to RT-PCR are 1,000, 200, 40, and 8 cells from left to right in each panel. Specific primers for CD4, IL-7Rα, and CD3-ε were used as positive and negative controls. (C) CD4+ cells were prepared with Dynabeads and placed in the chemotaxis chamber in duplicates. ELC and SLC maximally attracted CD4+ cells at 100–300 ng/ml. Results are expressed as the percentage of input cells migrating to the lower chamber containing BLC (1 μg/ml), ELC (300 ng/ml), or SLC (300 ng/ml). Data from individual wells are shown as filled diamonds and means as bars. The results are representative of three independent experiments. (D) Chemotactic response to BLC by CD4+ cells from intestine at E17.5 (filled circles) or B cells from Spl (open squares). Lines represent the averages of duplicated Transwells. The results are representative of at least three independent experiments.

Mentions: At E16.5, CD4+ cells begin to form segregated clusters in embryonic intestine, in contrast to the scattered pattern over the gut that is observed up to E15.5. By E17.5, increasing numbers of CD4+ cells are recruited to PP anlagen (Fig. 1 A). We attempted to investigate the molecular mechanisms underlying this clustering process. If the migration of CD4+ cells is regulated by specific chemokines, the corresponding chemokine receptors should be expressed in the PP inducers. Among the characterized receptors, CXCR5 and CCR7 have been suggested to participate in the accumulation of PP inducers. In particular, CXCR5 was a likely candidate, because mice genetically deficient in this gene possess few PPs 12. RT-PCR analyses revealed that both chemokine receptors are expressed in CD4+CD3−IL-7Rα1 cells sorted from the intestine (Fig. 1 B). Moreover, in vitro chemotactic analyses showed that their ligands, BLC, ELC, and SLC, all induced migration of CD4+CD3−IL-7Rα1 cells (Fig. 1 C). Furthermore, the magnitude of response of CD4+CD3−IL-7Rα1 cells to BLC was comparable to that observed for B lymphocytes (Fig. 1 D). It was likely that the overall higher frequency in CD4+CD3−IL-7Rα1 cells than in B cells reflected a high basal motility in CD4+CD3−IL-7Rα1 cells. Similar results were obtained with CD4+CD3−IL-7Rα1 cells from E15.5 intestines. These data show that PP inducers are highly competent to respond to chemokines.


Molecular basis for hematopoietic/mesenchymal interaction during initiation of Peyer's patch organogenesis.

Honda K, Nakano H, Yoshida H, Nishikawa S, Rennert P, Ikuta K, Tamechika M, Yamaguchi K, Fukumoto T, Chiba T, Nishikawa SI - J. Exp. Med. (2001)

Chemotactic activity of BLC, ELC, and SLC on CD4+CD3−IL-7Rα1 cells. (A) Whole-mount immunostaining analysis of E15.5, E16.5, and E17.5 gut with anti-CD4 mAb. (B) RT-PCR analysis on CXCR5 and CCR7 in FACS®-sorted CD4+CD3−IL-7Rα1 cells from intestine at E17.5. The numbers of cells subjected to RT-PCR are 1,000, 200, 40, and 8 cells from left to right in each panel. Specific primers for CD4, IL-7Rα, and CD3-ε were used as positive and negative controls. (C) CD4+ cells were prepared with Dynabeads and placed in the chemotaxis chamber in duplicates. ELC and SLC maximally attracted CD4+ cells at 100–300 ng/ml. Results are expressed as the percentage of input cells migrating to the lower chamber containing BLC (1 μg/ml), ELC (300 ng/ml), or SLC (300 ng/ml). Data from individual wells are shown as filled diamonds and means as bars. The results are representative of three independent experiments. (D) Chemotactic response to BLC by CD4+ cells from intestine at E17.5 (filled circles) or B cells from Spl (open squares). Lines represent the averages of duplicated Transwells. The results are representative of at least three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193398&req=5

Figure 1: Chemotactic activity of BLC, ELC, and SLC on CD4+CD3−IL-7Rα1 cells. (A) Whole-mount immunostaining analysis of E15.5, E16.5, and E17.5 gut with anti-CD4 mAb. (B) RT-PCR analysis on CXCR5 and CCR7 in FACS®-sorted CD4+CD3−IL-7Rα1 cells from intestine at E17.5. The numbers of cells subjected to RT-PCR are 1,000, 200, 40, and 8 cells from left to right in each panel. Specific primers for CD4, IL-7Rα, and CD3-ε were used as positive and negative controls. (C) CD4+ cells were prepared with Dynabeads and placed in the chemotaxis chamber in duplicates. ELC and SLC maximally attracted CD4+ cells at 100–300 ng/ml. Results are expressed as the percentage of input cells migrating to the lower chamber containing BLC (1 μg/ml), ELC (300 ng/ml), or SLC (300 ng/ml). Data from individual wells are shown as filled diamonds and means as bars. The results are representative of three independent experiments. (D) Chemotactic response to BLC by CD4+ cells from intestine at E17.5 (filled circles) or B cells from Spl (open squares). Lines represent the averages of duplicated Transwells. The results are representative of at least three independent experiments.
Mentions: At E16.5, CD4+ cells begin to form segregated clusters in embryonic intestine, in contrast to the scattered pattern over the gut that is observed up to E15.5. By E17.5, increasing numbers of CD4+ cells are recruited to PP anlagen (Fig. 1 A). We attempted to investigate the molecular mechanisms underlying this clustering process. If the migration of CD4+ cells is regulated by specific chemokines, the corresponding chemokine receptors should be expressed in the PP inducers. Among the characterized receptors, CXCR5 and CCR7 have been suggested to participate in the accumulation of PP inducers. In particular, CXCR5 was a likely candidate, because mice genetically deficient in this gene possess few PPs 12. RT-PCR analyses revealed that both chemokine receptors are expressed in CD4+CD3−IL-7Rα1 cells sorted from the intestine (Fig. 1 B). Moreover, in vitro chemotactic analyses showed that their ligands, BLC, ELC, and SLC, all induced migration of CD4+CD3−IL-7Rα1 cells (Fig. 1 C). Furthermore, the magnitude of response of CD4+CD3−IL-7Rα1 cells to BLC was comparable to that observed for B lymphocytes (Fig. 1 D). It was likely that the overall higher frequency in CD4+CD3−IL-7Rα1 cells than in B cells reflected a high basal motility in CD4+CD3−IL-7Rα1 cells. Similar results were obtained with CD4+CD3−IL-7Rα1 cells from E15.5 intestines. These data show that PP inducers are highly competent to respond to chemokines.

Bottom Line: The lymphoid population expresses CXCR5 and has a strong chemotactic response to B lymphocyte chemoattractant (BLC).Importantly, chemokines produced by VCAM-1(+)ICAM-1(+) nonlymphoid cells mediate the recruitment of lymphoid cells.Thus, promotion of PP development relies on mutual interaction between mesenchymal and lymphoid cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Syogoin-Kawaharacho 53, Sakyo-ku, Kyoto 606-8507, Japan. khonda@virus.kyoto-u.ac.jp

ABSTRACT
Mice deficient in lymphotoxin beta receptor (LTbetaR) or interleukin 7 receptor alpha (IL-7Ralpha) lack Peyer's patches (PPs). Deficiency in CXC chemokine receptor 5 (CXCR5) also severely affects the development of PPs. A molecular network involving these three signaling pathways has been implicated in PP organogenesis, but it remains unclear how they are connected during this process. We have shown that PP organogenesis is initiated at sites containing IL-7Ralpha(+) lymphoid cells and vascular cell adhesion molecule (VCAM)-1/intercellular adhesion molecule (ICAM)-1 expressing nonlymphoid elements. Here we characterize these lymphoid and nonlymphoid components in terms of chemokine signals. The lymphoid population expresses CXCR5 and has a strong chemotactic response to B lymphocyte chemoattractant (BLC). Importantly, chemokines produced by VCAM-1(+)ICAM-1(+) nonlymphoid cells mediate the recruitment of lymphoid cells. Furthermore, we show that these VCAM-1(+)ICAM-1(+) cells are mesenchymal cells that are activated by lymphoid cells through the LTbetaR to express adhesion molecules and chemokines. Thus, promotion of PP development relies on mutual interaction between mesenchymal and lymphoid cells.

Show MeSH
Related in: MedlinePlus