Limits...
Interleukin 18 acts on memory T helper cells type 1 to induce airway inflammation and hyperresponsiveness in a naive host mouse.

Sugimoto T, Ishikawa Y, Yoshimoto T, Hayashi N, Fujimoto J, Nakanishi K - J. Exp. Med. (2004)

Bottom Line: Thus, Th1 cells become harmful when they are stimulated with Ag and IL-18.Newly polarized Th1 cells and IFN-gamma-expressing Th1 cells, both of which express IL-18 receptor alpha chain strongly, produce IFN-gamma, IL-9, IL-13, granulocyte/macrophage colony-stimulating factor, tumor necrosis factor alpha, regulated on activation, normal T cell expressed and secreted, and macrophage inflammatory protein 1alpha upon stimulation with Ag, IL-2, and IL-18 in vitro.Thus, Ag and IL-18 stimulate memory Th1 cells to induce severe airway inflammation and AHR in the naive host.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Hyogo College of Medicine, Mukogawa-cho, Nishinomiya, 663-8501, Japan.

ABSTRACT
Interleukin (IL)-18 was originally regarded to induce T helper cell (Th)1-related cytokines. In general, factors favoring interferon (IFN)-gamma production are believed to abolish allergic diseases. Thus, we tested the role of IL-18 in regulation of bronchial asthma. To avoid a background response of host-derived T cells, we administered memory type Th1 or Th2 cells into unsensitized mice and examined their role in induction of bronchial asthma. Administration of antigen (Ag) induced both airway inflammation and airway hyperresponsiveness (AHR) in mice receiving memory Th2 cells. In contrast, the same treatment induced only airway inflammation but not AHR in mice receiving memory Th1 cells. However, these mice developed striking AHR when they were coadministered with IL-18. Furthermore, mice having received IFN-gamma-expressing Th1 cells sorted from polarized Th1 cells developed severe airway inflammation and AHR after intranasal administration of Ag and IL-18. Thus, Th1 cells become harmful when they are stimulated with Ag and IL-18. Newly polarized Th1 cells and IFN-gamma-expressing Th1 cells, both of which express IL-18 receptor alpha chain strongly, produce IFN-gamma, IL-9, IL-13, granulocyte/macrophage colony-stimulating factor, tumor necrosis factor alpha, regulated on activation, normal T cell expressed and secreted, and macrophage inflammatory protein 1alpha upon stimulation with Ag, IL-2, and IL-18 in vitro. Thus, Ag and IL-18 stimulate memory Th1 cells to induce severe airway inflammation and AHR in the naive host.

Show MeSH

Related in: MedlinePlus

IL-18 induced IFN-γ, IL-9, IL-13, GM-CSF, RANTES, and MIP-1α production from OVA-specific Th1 cells. (A) 105/ml naive splenic CD4+ CD62L+ T cells from DO11.10 Tg mice were cultured with 100 pM IL-2 plus 1 μM OVA323–339 and 106/ml irradiated T cell–depleted BALB/c splenocytes in six-well plates in a total 3-ml volume for 7 d. For differentiation of Th1 cells, 10 ng/ml IL-12 and 10 μg/ml anti–IL-4 were added to the culture. For the differentiation of Th2 cells, 1,000 U/ml IL-4, 10 μg/ml anti–IL-12p40, and 10 μg/ml anti–IFN-γ were added. Two rounds of antigenic stimulations under polarizing conditions were performed. After priming, cells were washed and recultured at 105/0.2 ml/well with 100 pM IL-2, 1 μM OVA323–339, plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes in the presence of various concentrations of IL-18 (0–100 ng/ml). After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-4, IL-5, IL-6, IL-9, IL-13, RANTES, MIP-1α, and GM-CSF contents by ELISA. Results are geometric means ± SEM. (B) Surface expression of IL-18Rα chain by flow cytometry. The percentages shown represent the proportion of IL-18Rα chain+ cells among CD4+ cells. (C) Polarized Th1 cells were cultured at 105/0.2 ml/well with medium alone or various combinations of 100 pM IL-2, 20 ng/ml IL-12, and 50 ng/ml IL-18 in the presence of 1 μM OVA323–339 plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes. After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-13, and GM-CSF contents by ELISA. Results are geometric means ± SEM.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2211833&req=5

fig3: IL-18 induced IFN-γ, IL-9, IL-13, GM-CSF, RANTES, and MIP-1α production from OVA-specific Th1 cells. (A) 105/ml naive splenic CD4+ CD62L+ T cells from DO11.10 Tg mice were cultured with 100 pM IL-2 plus 1 μM OVA323–339 and 106/ml irradiated T cell–depleted BALB/c splenocytes in six-well plates in a total 3-ml volume for 7 d. For differentiation of Th1 cells, 10 ng/ml IL-12 and 10 μg/ml anti–IL-4 were added to the culture. For the differentiation of Th2 cells, 1,000 U/ml IL-4, 10 μg/ml anti–IL-12p40, and 10 μg/ml anti–IFN-γ were added. Two rounds of antigenic stimulations under polarizing conditions were performed. After priming, cells were washed and recultured at 105/0.2 ml/well with 100 pM IL-2, 1 μM OVA323–339, plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes in the presence of various concentrations of IL-18 (0–100 ng/ml). After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-4, IL-5, IL-6, IL-9, IL-13, RANTES, MIP-1α, and GM-CSF contents by ELISA. Results are geometric means ± SEM. (B) Surface expression of IL-18Rα chain by flow cytometry. The percentages shown represent the proportion of IL-18Rα chain+ cells among CD4+ cells. (C) Polarized Th1 cells were cultured at 105/0.2 ml/well with medium alone or various combinations of 100 pM IL-2, 20 ng/ml IL-12, and 50 ng/ml IL-18 in the presence of 1 μM OVA323–339 plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes. After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-13, and GM-CSF contents by ELISA. Results are geometric means ± SEM.

Mentions: To understand how Ag and IL-18 induce airway inflammation and AHR, we next tested the types of cytokines or chemokines produced by Ag-, IL-2–, and IL-18–stimulated Th1 cells in vitro. We measured the concentrations of IL-4, IL-5, IL-6, IL-9, IL-13, TNF-α, GM-CSF, RANTES, eotaxin, MIP-1α, and IFN-γ in the culture supernatant of Th1 cells because they are reported to be deeply involved in induction of bronchial asthma (1–7). In particular, the chemokines RANTES, MIP-1α, and eotaxin are central to the delivery of eosinophils to the airway (53). Naive CD4+ T cells from OVA-specific TCR Tg mice were primed in vitro under Th1- or Th2-inducing conditions for two consecutive rounds of 7-d stimulation. Upon challenge with OVA peptide and IL-2, as reported elsewhere, Th2 cells produced significant amounts of IL-4, IL-5, IL-6, IL-9, and IL-13 but not IFN-γ, and Th1 cells produced IFN-γ but not Th2 cytokines (Fig. 3 A). Additional stimulation of Ag plus IL-2–stimulated Th1 cells with IL-18 dose dependently induced IFN-γ (Fig. 3 A; reference 44). Most surprisingly, this treatment simultaneously induced RANTES, MIP-1α, IL-9, IL-13, TNF-α (not depicted), and GM-CSF in Th1 cells (Fig. 3 A). Eotaxin was not detectable in any culture supernatant of Th1 cells. The mechanism underlying this difference in IL-18 responsiveness between Th1 and Th2 cells is principally explained by preferential expression of IL-18Rα chain on Th1 cells (44). As expected, Th1 cells express a high level of IL-18R, whereas Th2 cells express modestly (Fig. 3 B). Furthermore, they simultaneously produced a large amount of chemokines (Fig. 3 A). Thus, IL-18R–expressing Th1 cells are target cells for IL-18 and responded to it not only by increased production of IFN-γ, but also by production of some Th2 cytokines (IL-9 and IL-13), TNF-α (not depicted), GM-CSF, and chemokines. Because IL-12 plus IL-18 is known to induce IFN-γ production from Th1 cells but inhibit IL-4 and IL-13 production from Th2 cells in vivo and in vitro (30, 33, 38, 44), it is important to test the effect of IL-12 on the ability of IL-18 to induce IL-13 from Th1 cells. We found that IL-12 slightly diminished the production of IL-13 and GM-CSF from Th1 cells stimulated with IL-18 alone or IL-18 plus IL-2 in the presence of OVA (Fig. 3 C). Moreover, additional IL-12 stimulation did not affect IL-13 and GM-CSF production from Th1 cells stimulated with immobilized anti-CD3 plus anti-CD28 antibodies (not depicted). Thus, IL-12 is essential for induction of Th1 cells but not critically involved in induction of cytokine production from Th1 cells. As reported elsewhere, Th2 cells failed to respond to IL-18 by enhanced production of IL-4, although they modestly increased production of IL-5, IL-9, and IL-13 in response to IL-18 (Fig. 3 A). These results suggested the possible involvement of cytokines, particularly of IFN-γ, TNF-α, IL-9, IL-13, GM-CSF, and some chemokines from Th1 cells in induction of both severe airway inflammation and AHR.


Interleukin 18 acts on memory T helper cells type 1 to induce airway inflammation and hyperresponsiveness in a naive host mouse.

Sugimoto T, Ishikawa Y, Yoshimoto T, Hayashi N, Fujimoto J, Nakanishi K - J. Exp. Med. (2004)

IL-18 induced IFN-γ, IL-9, IL-13, GM-CSF, RANTES, and MIP-1α production from OVA-specific Th1 cells. (A) 105/ml naive splenic CD4+ CD62L+ T cells from DO11.10 Tg mice were cultured with 100 pM IL-2 plus 1 μM OVA323–339 and 106/ml irradiated T cell–depleted BALB/c splenocytes in six-well plates in a total 3-ml volume for 7 d. For differentiation of Th1 cells, 10 ng/ml IL-12 and 10 μg/ml anti–IL-4 were added to the culture. For the differentiation of Th2 cells, 1,000 U/ml IL-4, 10 μg/ml anti–IL-12p40, and 10 μg/ml anti–IFN-γ were added. Two rounds of antigenic stimulations under polarizing conditions were performed. After priming, cells were washed and recultured at 105/0.2 ml/well with 100 pM IL-2, 1 μM OVA323–339, plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes in the presence of various concentrations of IL-18 (0–100 ng/ml). After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-4, IL-5, IL-6, IL-9, IL-13, RANTES, MIP-1α, and GM-CSF contents by ELISA. Results are geometric means ± SEM. (B) Surface expression of IL-18Rα chain by flow cytometry. The percentages shown represent the proportion of IL-18Rα chain+ cells among CD4+ cells. (C) Polarized Th1 cells were cultured at 105/0.2 ml/well with medium alone or various combinations of 100 pM IL-2, 20 ng/ml IL-12, and 50 ng/ml IL-18 in the presence of 1 μM OVA323–339 plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes. After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-13, and GM-CSF contents by ELISA. Results are geometric means ± SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211833&req=5

fig3: IL-18 induced IFN-γ, IL-9, IL-13, GM-CSF, RANTES, and MIP-1α production from OVA-specific Th1 cells. (A) 105/ml naive splenic CD4+ CD62L+ T cells from DO11.10 Tg mice were cultured with 100 pM IL-2 plus 1 μM OVA323–339 and 106/ml irradiated T cell–depleted BALB/c splenocytes in six-well plates in a total 3-ml volume for 7 d. For differentiation of Th1 cells, 10 ng/ml IL-12 and 10 μg/ml anti–IL-4 were added to the culture. For the differentiation of Th2 cells, 1,000 U/ml IL-4, 10 μg/ml anti–IL-12p40, and 10 μg/ml anti–IFN-γ were added. Two rounds of antigenic stimulations under polarizing conditions were performed. After priming, cells were washed and recultured at 105/0.2 ml/well with 100 pM IL-2, 1 μM OVA323–339, plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes in the presence of various concentrations of IL-18 (0–100 ng/ml). After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-4, IL-5, IL-6, IL-9, IL-13, RANTES, MIP-1α, and GM-CSF contents by ELISA. Results are geometric means ± SEM. (B) Surface expression of IL-18Rα chain by flow cytometry. The percentages shown represent the proportion of IL-18Rα chain+ cells among CD4+ cells. (C) Polarized Th1 cells were cultured at 105/0.2 ml/well with medium alone or various combinations of 100 pM IL-2, 20 ng/ml IL-12, and 50 ng/ml IL-18 in the presence of 1 μM OVA323–339 plus 105/0.2 ml irradiated T cell–depleted BALB/c splenocytes. After 48 h of culture, supernatants were harvested and tested for IFN-γ, IL-13, and GM-CSF contents by ELISA. Results are geometric means ± SEM.
Mentions: To understand how Ag and IL-18 induce airway inflammation and AHR, we next tested the types of cytokines or chemokines produced by Ag-, IL-2–, and IL-18–stimulated Th1 cells in vitro. We measured the concentrations of IL-4, IL-5, IL-6, IL-9, IL-13, TNF-α, GM-CSF, RANTES, eotaxin, MIP-1α, and IFN-γ in the culture supernatant of Th1 cells because they are reported to be deeply involved in induction of bronchial asthma (1–7). In particular, the chemokines RANTES, MIP-1α, and eotaxin are central to the delivery of eosinophils to the airway (53). Naive CD4+ T cells from OVA-specific TCR Tg mice were primed in vitro under Th1- or Th2-inducing conditions for two consecutive rounds of 7-d stimulation. Upon challenge with OVA peptide and IL-2, as reported elsewhere, Th2 cells produced significant amounts of IL-4, IL-5, IL-6, IL-9, and IL-13 but not IFN-γ, and Th1 cells produced IFN-γ but not Th2 cytokines (Fig. 3 A). Additional stimulation of Ag plus IL-2–stimulated Th1 cells with IL-18 dose dependently induced IFN-γ (Fig. 3 A; reference 44). Most surprisingly, this treatment simultaneously induced RANTES, MIP-1α, IL-9, IL-13, TNF-α (not depicted), and GM-CSF in Th1 cells (Fig. 3 A). Eotaxin was not detectable in any culture supernatant of Th1 cells. The mechanism underlying this difference in IL-18 responsiveness between Th1 and Th2 cells is principally explained by preferential expression of IL-18Rα chain on Th1 cells (44). As expected, Th1 cells express a high level of IL-18R, whereas Th2 cells express modestly (Fig. 3 B). Furthermore, they simultaneously produced a large amount of chemokines (Fig. 3 A). Thus, IL-18R–expressing Th1 cells are target cells for IL-18 and responded to it not only by increased production of IFN-γ, but also by production of some Th2 cytokines (IL-9 and IL-13), TNF-α (not depicted), GM-CSF, and chemokines. Because IL-12 plus IL-18 is known to induce IFN-γ production from Th1 cells but inhibit IL-4 and IL-13 production from Th2 cells in vivo and in vitro (30, 33, 38, 44), it is important to test the effect of IL-12 on the ability of IL-18 to induce IL-13 from Th1 cells. We found that IL-12 slightly diminished the production of IL-13 and GM-CSF from Th1 cells stimulated with IL-18 alone or IL-18 plus IL-2 in the presence of OVA (Fig. 3 C). Moreover, additional IL-12 stimulation did not affect IL-13 and GM-CSF production from Th1 cells stimulated with immobilized anti-CD3 plus anti-CD28 antibodies (not depicted). Thus, IL-12 is essential for induction of Th1 cells but not critically involved in induction of cytokine production from Th1 cells. As reported elsewhere, Th2 cells failed to respond to IL-18 by enhanced production of IL-4, although they modestly increased production of IL-5, IL-9, and IL-13 in response to IL-18 (Fig. 3 A). These results suggested the possible involvement of cytokines, particularly of IFN-γ, TNF-α, IL-9, IL-13, GM-CSF, and some chemokines from Th1 cells in induction of both severe airway inflammation and AHR.

Bottom Line: Thus, Th1 cells become harmful when they are stimulated with Ag and IL-18.Newly polarized Th1 cells and IFN-gamma-expressing Th1 cells, both of which express IL-18 receptor alpha chain strongly, produce IFN-gamma, IL-9, IL-13, granulocyte/macrophage colony-stimulating factor, tumor necrosis factor alpha, regulated on activation, normal T cell expressed and secreted, and macrophage inflammatory protein 1alpha upon stimulation with Ag, IL-2, and IL-18 in vitro.Thus, Ag and IL-18 stimulate memory Th1 cells to induce severe airway inflammation and AHR in the naive host.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Hyogo College of Medicine, Mukogawa-cho, Nishinomiya, 663-8501, Japan.

ABSTRACT
Interleukin (IL)-18 was originally regarded to induce T helper cell (Th)1-related cytokines. In general, factors favoring interferon (IFN)-gamma production are believed to abolish allergic diseases. Thus, we tested the role of IL-18 in regulation of bronchial asthma. To avoid a background response of host-derived T cells, we administered memory type Th1 or Th2 cells into unsensitized mice and examined their role in induction of bronchial asthma. Administration of antigen (Ag) induced both airway inflammation and airway hyperresponsiveness (AHR) in mice receiving memory Th2 cells. In contrast, the same treatment induced only airway inflammation but not AHR in mice receiving memory Th1 cells. However, these mice developed striking AHR when they were coadministered with IL-18. Furthermore, mice having received IFN-gamma-expressing Th1 cells sorted from polarized Th1 cells developed severe airway inflammation and AHR after intranasal administration of Ag and IL-18. Thus, Th1 cells become harmful when they are stimulated with Ag and IL-18. Newly polarized Th1 cells and IFN-gamma-expressing Th1 cells, both of which express IL-18 receptor alpha chain strongly, produce IFN-gamma, IL-9, IL-13, granulocyte/macrophage colony-stimulating factor, tumor necrosis factor alpha, regulated on activation, normal T cell expressed and secreted, and macrophage inflammatory protein 1alpha upon stimulation with Ag, IL-2, and IL-18 in vitro. Thus, Ag and IL-18 stimulate memory Th1 cells to induce severe airway inflammation and AHR in the naive host.

Show MeSH
Related in: MedlinePlus