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Induction of airway mucus production By T helper 2 (Th2) cells: a critical role for interleukin 4 in cell recruitment but not mucus production.

Cohn L, Homer RJ, Marinov A, Rankin J, Bottomly K - J. Exp. Med. (1997)

Bottom Line: When this defect in homing was overcome by administration of TNF-alpha, IL-4 -/- Th2 cells induced mucus as effectively as IL-4 +/+ Th2 cells.These studies establish a role for Th2 cells in mucus production and dissect the effector functions of IL-4 in these processes.These data suggest that IL-4 is crucial for Th2 cell recruitment to the lung and for induction of inflammation, but has no direct role in mucus production.

View Article: PubMed Central - PubMed

Affiliation: Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA. lcohn@biomed.med.yale.edu

ABSTRACT
Airway inflammation is believed to stimulate mucus production in asthmatic patients. Increased mucus secretion is an important clinical symptom and contributes to airway obstruction in asthma. Activated CD4 Th1 and Th2 cells have both been identified in airway biopsies of asthmatics but their role in mucus production is not clear. Using CD4 T cells from mice transgenic for the OVA-specific TCR, we studied the role of Th1 and Th2 cells in airway inflammation and mucus production. Airway inflammation induced by Th2 cells was comprised of eosinophils and lymphocytes; features found in asthmatic patients. Additionally, there was a marked increase in mucus production in mice that received Th2 cells and inhaled OVA, but not in mice that received Th1 cells. However, OVA-specific Th2 cells from IL-4-deficient mice were not recruited to the lung and did not induce mucus production. When this defect in homing was overcome by administration of TNF-alpha, IL-4 -/- Th2 cells induced mucus as effectively as IL-4 +/+ Th2 cells. These studies establish a role for Th2 cells in mucus production and dissect the effector functions of IL-4 in these processes. These data suggest that IL-4 is crucial for Th2 cell recruitment to the lung and for induction of inflammation, but has no direct role in mucus production.

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Cytokine production by Th1 or Th2-like cells before and after transfer. (A) At the  time of transfer into recipient  mice in vitro generated  DO11.10 CD4 Th1, Th2 or  freshly isolated CD4 T cells from  naive BALB/c mice (N) were  cultured with antigen presenting  cells (2 × 106 cells/ml) in the  presence of pOVA323-339. (B) After 7 d of exposure to inhaled  OVA, BAL was performed on  individual mice that received  Th1 (Th1-OVA), Th2 (Th2-OVA), or naive (N) CD4 T  cells. Total leukocytes recovered  from BAL were restimulated in vitro with pOVA323-339. BAL cells from mice that received naive CD4 T cells (N) and inhaled OVA contained <3% lymphocytes and were insufficient for cytokine analysis. Supernatants were collected at 48 h and cytokine ELISAs were performed. Cytokines production in  BAL was adjusted for 106 DO11.10 CD4 T cells per ml as determined by FACS® analysis. Mean cytokine levels (±SEM) are shown (n = 4–5 mice per  group). One experiment is shown and is representative of three experiments.
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Figure 1: Cytokine production by Th1 or Th2-like cells before and after transfer. (A) At the time of transfer into recipient mice in vitro generated DO11.10 CD4 Th1, Th2 or freshly isolated CD4 T cells from naive BALB/c mice (N) were cultured with antigen presenting cells (2 × 106 cells/ml) in the presence of pOVA323-339. (B) After 7 d of exposure to inhaled OVA, BAL was performed on individual mice that received Th1 (Th1-OVA), Th2 (Th2-OVA), or naive (N) CD4 T cells. Total leukocytes recovered from BAL were restimulated in vitro with pOVA323-339. BAL cells from mice that received naive CD4 T cells (N) and inhaled OVA contained <3% lymphocytes and were insufficient for cytokine analysis. Supernatants were collected at 48 h and cytokine ELISAs were performed. Cytokines production in BAL was adjusted for 106 DO11.10 CD4 T cells per ml as determined by FACS® analysis. Mean cytokine levels (±SEM) are shown (n = 4–5 mice per group). One experiment is shown and is representative of three experiments.

Mentions: To investigate the effects of different T cell subsets on airway pathology, Th1 and Th2 cells were generated from CD4 T cells isolated from DO11.10 mice which are transgenic for the TCR recognizing pOVA 323-339. Using a standard procedure to polarize CD4 T cell responses (41), splenic CD4 T cells were stimulated by pOVA323-339 in the presence of IL-12, IL-2, and anti–IL-4 to induce Th1-like cells or IL-4, IL-2, and anti–IFN-γ to induce Th2-like cells. 99% of the resulting activated Th1 or Th2 effector cells expressed CD4 and the DO11.10 TCR, recognized by the clonotypic monoclonal antibody, KJ1-26. An aliquot of cells from each culture was restimulated in vitro in the presence of pOVA323-339 and APCs, and supernatants were assayed for IFN-γ, IL-4, IL-5 (Fig. 1 A), and IL-10 (data not shown). The CD4 T cells stimulated to differentiate into Th1 cells produced high levels of IFN-γ and undetectable IL-4, IL-5, and low levels of IL-10, while the cells stimulated to differentiate into Th2 cells secreted high levels of IL-4, IL-5, IL-10, and minimal IFN-γ. Thus, we generated CD4 Th1 and Th2 effector cells responsive to OVA peptide. Naive CD4 T cells (N) freshly isolated from BALB/c mice did not secrete cytokines in response to OVA.


Induction of airway mucus production By T helper 2 (Th2) cells: a critical role for interleukin 4 in cell recruitment but not mucus production.

Cohn L, Homer RJ, Marinov A, Rankin J, Bottomly K - J. Exp. Med. (1997)

Cytokine production by Th1 or Th2-like cells before and after transfer. (A) At the  time of transfer into recipient  mice in vitro generated  DO11.10 CD4 Th1, Th2 or  freshly isolated CD4 T cells from  naive BALB/c mice (N) were  cultured with antigen presenting  cells (2 × 106 cells/ml) in the  presence of pOVA323-339. (B) After 7 d of exposure to inhaled  OVA, BAL was performed on  individual mice that received  Th1 (Th1-OVA), Th2 (Th2-OVA), or naive (N) CD4 T  cells. Total leukocytes recovered  from BAL were restimulated in vitro with pOVA323-339. BAL cells from mice that received naive CD4 T cells (N) and inhaled OVA contained <3% lymphocytes and were insufficient for cytokine analysis. Supernatants were collected at 48 h and cytokine ELISAs were performed. Cytokines production in  BAL was adjusted for 106 DO11.10 CD4 T cells per ml as determined by FACS® analysis. Mean cytokine levels (±SEM) are shown (n = 4–5 mice per  group). One experiment is shown and is representative of three experiments.
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Related In: Results  -  Collection

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Figure 1: Cytokine production by Th1 or Th2-like cells before and after transfer. (A) At the time of transfer into recipient mice in vitro generated DO11.10 CD4 Th1, Th2 or freshly isolated CD4 T cells from naive BALB/c mice (N) were cultured with antigen presenting cells (2 × 106 cells/ml) in the presence of pOVA323-339. (B) After 7 d of exposure to inhaled OVA, BAL was performed on individual mice that received Th1 (Th1-OVA), Th2 (Th2-OVA), or naive (N) CD4 T cells. Total leukocytes recovered from BAL were restimulated in vitro with pOVA323-339. BAL cells from mice that received naive CD4 T cells (N) and inhaled OVA contained <3% lymphocytes and were insufficient for cytokine analysis. Supernatants were collected at 48 h and cytokine ELISAs were performed. Cytokines production in BAL was adjusted for 106 DO11.10 CD4 T cells per ml as determined by FACS® analysis. Mean cytokine levels (±SEM) are shown (n = 4–5 mice per group). One experiment is shown and is representative of three experiments.
Mentions: To investigate the effects of different T cell subsets on airway pathology, Th1 and Th2 cells were generated from CD4 T cells isolated from DO11.10 mice which are transgenic for the TCR recognizing pOVA 323-339. Using a standard procedure to polarize CD4 T cell responses (41), splenic CD4 T cells were stimulated by pOVA323-339 in the presence of IL-12, IL-2, and anti–IL-4 to induce Th1-like cells or IL-4, IL-2, and anti–IFN-γ to induce Th2-like cells. 99% of the resulting activated Th1 or Th2 effector cells expressed CD4 and the DO11.10 TCR, recognized by the clonotypic monoclonal antibody, KJ1-26. An aliquot of cells from each culture was restimulated in vitro in the presence of pOVA323-339 and APCs, and supernatants were assayed for IFN-γ, IL-4, IL-5 (Fig. 1 A), and IL-10 (data not shown). The CD4 T cells stimulated to differentiate into Th1 cells produced high levels of IFN-γ and undetectable IL-4, IL-5, and low levels of IL-10, while the cells stimulated to differentiate into Th2 cells secreted high levels of IL-4, IL-5, IL-10, and minimal IFN-γ. Thus, we generated CD4 Th1 and Th2 effector cells responsive to OVA peptide. Naive CD4 T cells (N) freshly isolated from BALB/c mice did not secrete cytokines in response to OVA.

Bottom Line: When this defect in homing was overcome by administration of TNF-alpha, IL-4 -/- Th2 cells induced mucus as effectively as IL-4 +/+ Th2 cells.These studies establish a role for Th2 cells in mucus production and dissect the effector functions of IL-4 in these processes.These data suggest that IL-4 is crucial for Th2 cell recruitment to the lung and for induction of inflammation, but has no direct role in mucus production.

View Article: PubMed Central - PubMed

Affiliation: Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA. lcohn@biomed.med.yale.edu

ABSTRACT
Airway inflammation is believed to stimulate mucus production in asthmatic patients. Increased mucus secretion is an important clinical symptom and contributes to airway obstruction in asthma. Activated CD4 Th1 and Th2 cells have both been identified in airway biopsies of asthmatics but their role in mucus production is not clear. Using CD4 T cells from mice transgenic for the OVA-specific TCR, we studied the role of Th1 and Th2 cells in airway inflammation and mucus production. Airway inflammation induced by Th2 cells was comprised of eosinophils and lymphocytes; features found in asthmatic patients. Additionally, there was a marked increase in mucus production in mice that received Th2 cells and inhaled OVA, but not in mice that received Th1 cells. However, OVA-specific Th2 cells from IL-4-deficient mice were not recruited to the lung and did not induce mucus production. When this defect in homing was overcome by administration of TNF-alpha, IL-4 -/- Th2 cells induced mucus as effectively as IL-4 +/+ Th2 cells. These studies establish a role for Th2 cells in mucus production and dissect the effector functions of IL-4 in these processes. These data suggest that IL-4 is crucial for Th2 cell recruitment to the lung and for induction of inflammation, but has no direct role in mucus production.

Show MeSH
Related in: MedlinePlus