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CC chemokine receptor (CCR)3/eotaxin is followed by CCR4/monocyte-derived chemokine in mediating pulmonary T helper lymphocyte type 2 recruitment after serial antigen challenge in vivo.

Lloyd CM, Delaney T, Nguyen T, Tian J, Martinez-A C, Coyle AJ, Gutierrez-Ramos JC - J. Exp. Med. (2000)

Bottom Line: Tracking of these cells after repeated antigen challenge has established that both CCR3/eotaxin and CCR4/MDC axes contribute to the recruitment of Th2 cells to the lung, demonstrating the in vivo relevance of the expression of these receptors on Th2 cells.We have shown that involvement of the CCR3/eotaxin pathway is confined to early stages of the response in vivo, whereas repeated antigen stimulation results in the predominant use of the CCR4/MDC pathway.We propose that effector Th2 cells respond to both CCR3/eotaxin and CCR4/MDC pathways initially, but that a progressive increase in CCR4-positive cells results in the predominance of the CCR4/MDC axis in the long-term recruitment of Th2 cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts 02139, USA. c.lloyd@ic.ac.uk

ABSTRACT
Isolated peripheral blood CD4 cells from allergic individuals express CC chemokine receptor (CCR)3 and CCR4 after expansion in vitro. In addition, human T helper type 2 (Th2) cells polarized in vitro selectively express CCR3 and CCR4 at certain stages of activation/differentiation and respond preferentially to the ligands eotaxin and monocyte-derived chemokine (MDC). However, controversy arises when the in vivo significance of this distinct expression is discussed. To address the functional role of CCR3/eotaxin and CCR4/MDC during the in vivo recruitment of Th2 cells, we have transferred effector Th cells into naive mice to induce allergic airway disease. Tracking of these cells after repeated antigen challenge has established that both CCR3/eotaxin and CCR4/MDC axes contribute to the recruitment of Th2 cells to the lung, demonstrating the in vivo relevance of the expression of these receptors on Th2 cells. We have shown that involvement of the CCR3/eotaxin pathway is confined to early stages of the response in vivo, whereas repeated antigen stimulation results in the predominant use of the CCR4/MDC pathway. We propose that effector Th2 cells respond to both CCR3/eotaxin and CCR4/MDC pathways initially, but that a progressive increase in CCR4-positive cells results in the predominance of the CCR4/MDC axis in the long-term recruitment of Th2 cells in vivo.

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Adoptive transfer of Th effector cells leads to AAD. BALB/c mice were injected with 2 × 106 cells followed by daily aerosolized OVA. Mice were killed on day 4 or 7 for analysis. Results shown are from two different experiments with five mice per group for each experiment. (A, i) Total leukocytes were recovered from BAL, counted, and cytospun, and differential counts were calculated. Total numbers of eosinophils (black bars) and neutrophils (white bars) are shown for days 4 (left) and 7 (right) after transfer of Th1 (gray background throughout figure) or Th2 cells (white background throughout figure). (A, ii) The percentage of eosinophils (black bars) and neutrophils (white bars) within peribronchiolar infiltrates was counted on days 4 (left) and 7 (right) after transfer of either Th1 or Th2 cells. (A, iii) Representative sections from mice transferred with Th1 cells (a and c) or Th2 cells (b and d) stained with eosinophil peroxidase (a and b) or chloroacetate esterase (c and d) to show the presence of eosinophils or neutrophils, respectively (original magnifications: ×400; [inset] ×1,000). (A, iv) BHR was measured after provocation with 20 mg/ml methacholine at days 4 and 7. Bars show mean Penh (± SEM) of groups of five mice before (gray bars) or after (white bars) methacholine stimulation. (B) BAL cytokines were measured by ELISA in OVA- (gray bars) and PBS-challenged mice (white bars) after transfer of Th1 (gray panel) or Th2 cells (white panel). Each bar represents the mean (± SEM) concentration in groups of five mice on day 4.
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Figure 1: Adoptive transfer of Th effector cells leads to AAD. BALB/c mice were injected with 2 × 106 cells followed by daily aerosolized OVA. Mice were killed on day 4 or 7 for analysis. Results shown are from two different experiments with five mice per group for each experiment. (A, i) Total leukocytes were recovered from BAL, counted, and cytospun, and differential counts were calculated. Total numbers of eosinophils (black bars) and neutrophils (white bars) are shown for days 4 (left) and 7 (right) after transfer of Th1 (gray background throughout figure) or Th2 cells (white background throughout figure). (A, ii) The percentage of eosinophils (black bars) and neutrophils (white bars) within peribronchiolar infiltrates was counted on days 4 (left) and 7 (right) after transfer of either Th1 or Th2 cells. (A, iii) Representative sections from mice transferred with Th1 cells (a and c) or Th2 cells (b and d) stained with eosinophil peroxidase (a and b) or chloroacetate esterase (c and d) to show the presence of eosinophils or neutrophils, respectively (original magnifications: ×400; [inset] ×1,000). (A, iv) BHR was measured after provocation with 20 mg/ml methacholine at days 4 and 7. Bars show mean Penh (± SEM) of groups of five mice before (gray bars) or after (white bars) methacholine stimulation. (B) BAL cytokines were measured by ELISA in OVA- (gray bars) and PBS-challenged mice (white bars) after transfer of Th1 (gray panel) or Th2 cells (white panel). Each bar represents the mean (± SEM) concentration in groups of five mice on day 4.

Mentions: For blocking studies, mice were injected daily with 100 μg per mouse polyclonal rabbit antieotaxin Abs 7 or polyclonal rabbit anti–murine MDC 13 30 min before OVA challenge. Mice were then killed at day 4 after T cell transfer (after three antigen challenges) or at day 7 (after six antigen challenges), as shown in Fig. 1.


CC chemokine receptor (CCR)3/eotaxin is followed by CCR4/monocyte-derived chemokine in mediating pulmonary T helper lymphocyte type 2 recruitment after serial antigen challenge in vivo.

Lloyd CM, Delaney T, Nguyen T, Tian J, Martinez-A C, Coyle AJ, Gutierrez-Ramos JC - J. Exp. Med. (2000)

Adoptive transfer of Th effector cells leads to AAD. BALB/c mice were injected with 2 × 106 cells followed by daily aerosolized OVA. Mice were killed on day 4 or 7 for analysis. Results shown are from two different experiments with five mice per group for each experiment. (A, i) Total leukocytes were recovered from BAL, counted, and cytospun, and differential counts were calculated. Total numbers of eosinophils (black bars) and neutrophils (white bars) are shown for days 4 (left) and 7 (right) after transfer of Th1 (gray background throughout figure) or Th2 cells (white background throughout figure). (A, ii) The percentage of eosinophils (black bars) and neutrophils (white bars) within peribronchiolar infiltrates was counted on days 4 (left) and 7 (right) after transfer of either Th1 or Th2 cells. (A, iii) Representative sections from mice transferred with Th1 cells (a and c) or Th2 cells (b and d) stained with eosinophil peroxidase (a and b) or chloroacetate esterase (c and d) to show the presence of eosinophils or neutrophils, respectively (original magnifications: ×400; [inset] ×1,000). (A, iv) BHR was measured after provocation with 20 mg/ml methacholine at days 4 and 7. Bars show mean Penh (± SEM) of groups of five mice before (gray bars) or after (white bars) methacholine stimulation. (B) BAL cytokines were measured by ELISA in OVA- (gray bars) and PBS-challenged mice (white bars) after transfer of Th1 (gray panel) or Th2 cells (white panel). Each bar represents the mean (± SEM) concentration in groups of five mice on day 4.
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Related In: Results  -  Collection

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Figure 1: Adoptive transfer of Th effector cells leads to AAD. BALB/c mice were injected with 2 × 106 cells followed by daily aerosolized OVA. Mice were killed on day 4 or 7 for analysis. Results shown are from two different experiments with five mice per group for each experiment. (A, i) Total leukocytes were recovered from BAL, counted, and cytospun, and differential counts were calculated. Total numbers of eosinophils (black bars) and neutrophils (white bars) are shown for days 4 (left) and 7 (right) after transfer of Th1 (gray background throughout figure) or Th2 cells (white background throughout figure). (A, ii) The percentage of eosinophils (black bars) and neutrophils (white bars) within peribronchiolar infiltrates was counted on days 4 (left) and 7 (right) after transfer of either Th1 or Th2 cells. (A, iii) Representative sections from mice transferred with Th1 cells (a and c) or Th2 cells (b and d) stained with eosinophil peroxidase (a and b) or chloroacetate esterase (c and d) to show the presence of eosinophils or neutrophils, respectively (original magnifications: ×400; [inset] ×1,000). (A, iv) BHR was measured after provocation with 20 mg/ml methacholine at days 4 and 7. Bars show mean Penh (± SEM) of groups of five mice before (gray bars) or after (white bars) methacholine stimulation. (B) BAL cytokines were measured by ELISA in OVA- (gray bars) and PBS-challenged mice (white bars) after transfer of Th1 (gray panel) or Th2 cells (white panel). Each bar represents the mean (± SEM) concentration in groups of five mice on day 4.
Mentions: For blocking studies, mice were injected daily with 100 μg per mouse polyclonal rabbit antieotaxin Abs 7 or polyclonal rabbit anti–murine MDC 13 30 min before OVA challenge. Mice were then killed at day 4 after T cell transfer (after three antigen challenges) or at day 7 (after six antigen challenges), as shown in Fig. 1.

Bottom Line: Tracking of these cells after repeated antigen challenge has established that both CCR3/eotaxin and CCR4/MDC axes contribute to the recruitment of Th2 cells to the lung, demonstrating the in vivo relevance of the expression of these receptors on Th2 cells.We have shown that involvement of the CCR3/eotaxin pathway is confined to early stages of the response in vivo, whereas repeated antigen stimulation results in the predominant use of the CCR4/MDC pathway.We propose that effector Th2 cells respond to both CCR3/eotaxin and CCR4/MDC pathways initially, but that a progressive increase in CCR4-positive cells results in the predominance of the CCR4/MDC axis in the long-term recruitment of Th2 cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts 02139, USA. c.lloyd@ic.ac.uk

ABSTRACT
Isolated peripheral blood CD4 cells from allergic individuals express CC chemokine receptor (CCR)3 and CCR4 after expansion in vitro. In addition, human T helper type 2 (Th2) cells polarized in vitro selectively express CCR3 and CCR4 at certain stages of activation/differentiation and respond preferentially to the ligands eotaxin and monocyte-derived chemokine (MDC). However, controversy arises when the in vivo significance of this distinct expression is discussed. To address the functional role of CCR3/eotaxin and CCR4/MDC during the in vivo recruitment of Th2 cells, we have transferred effector Th cells into naive mice to induce allergic airway disease. Tracking of these cells after repeated antigen challenge has established that both CCR3/eotaxin and CCR4/MDC axes contribute to the recruitment of Th2 cells to the lung, demonstrating the in vivo relevance of the expression of these receptors on Th2 cells. We have shown that involvement of the CCR3/eotaxin pathway is confined to early stages of the response in vivo, whereas repeated antigen stimulation results in the predominant use of the CCR4/MDC pathway. We propose that effector Th2 cells respond to both CCR3/eotaxin and CCR4/MDC pathways initially, but that a progressive increase in CCR4-positive cells results in the predominance of the CCR4/MDC axis in the long-term recruitment of Th2 cells in vivo.

Show MeSH
Related in: MedlinePlus