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Attenuation of lung inflammation and fibrosis in CD69-deficient mice after intratracheal bleomycin.

Yamauchi K, Kasuya Y, Kuroda F, Tanaka K, Tsuyusaki J, Ishizaki S, Matsunaga H, Iwamura C, Nakayama T, Tatsumi K - Respir. Res. (2011)

Bottom Line: Cluster of differentiation 69 (CD69), an early activation marker antigen on T and B cells, is also expressed on activated macrophages and neutrophils, suggesting that CD69 may play a role in inflammatory diseases.Lung inflammation in the acute phase (7 dpi) was investigated by differential cell counts and cytokine array analyses of bronchoalveolar lavage fluid.In addition, lung fibrotic changes were evaluated at 14 dpi by histopathology and collagen assays.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Respirology, Graduate School of Medicine, Chiba University, Chiba, Japan.

ABSTRACT

Background: Cluster of differentiation 69 (CD69), an early activation marker antigen on T and B cells, is also expressed on activated macrophages and neutrophils, suggesting that CD69 may play a role in inflammatory diseases. To determine the effect of CD69 deficiency on bleomycin(BLM)-induced lung injury, we evaluated the inflammatory response following intratracheal BLM administration and the subsequent fibrotic changes in wild type (WT) and CD69-deficient (CD69-/-) mice.

Methods: The mice received a single dose of 3 mg/kg body weight of BLM and were sacrificed at 7 or 14 days post-instillation (dpi). Lung inflammation in the acute phase (7 dpi) was investigated by differential cell counts and cytokine array analyses of bronchoalveolar lavage fluid. In addition, lung fibrotic changes were evaluated at 14 dpi by histopathology and collagen assays. We also used reverse transcription polymerase chain reaction to measure the mRNA expression level of transforming growth factor β1 (TGF-β1) in the lungs of BLM-treated mice.

Results: CD69-/- mice exhibited less lung damage than WT mice, as shown by reductions in the following indices: (1) loss of body weight, (2) wet/dry ratio of lung, (3) cytokine levels in BALF, (4) histological evidence of lung injury, (5) lung collagen deposition, and (6) TGF-β1 mRNA expression in the lung.

Conclusion: The present study clearly demonstrates that CD69 plays an important role in the progression of lung injury induced by BLM.

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Effect of bleomycin treatment on cytokine expression in wild-type and CD69-deficient mice. (A) Cytokine array analyses of BALF 7 d after the instillation of BLM or PBS (sham treatment) in WT or CD69-/- mice. Cytokines with increased expression levels are boxed. (B) Expression levels of these cytokines in WT (black bars) and CD69-/- (white bars) mice. Each expression level was normalized by that of the positive control. The stimulation index is the ratio of the expression level of a cytokine in BLM-treated mice to that in sham-treated mice. Results are expressed as the mean (SEM) (n = 4 mice per group). *P < 0.05, **P < 0.01.
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Figure 4: Effect of bleomycin treatment on cytokine expression in wild-type and CD69-deficient mice. (A) Cytokine array analyses of BALF 7 d after the instillation of BLM or PBS (sham treatment) in WT or CD69-/- mice. Cytokines with increased expression levels are boxed. (B) Expression levels of these cytokines in WT (black bars) and CD69-/- (white bars) mice. Each expression level was normalized by that of the positive control. The stimulation index is the ratio of the expression level of a cytokine in BLM-treated mice to that in sham-treated mice. Results are expressed as the mean (SEM) (n = 4 mice per group). *P < 0.05, **P < 0.01.

Mentions: To evaluate whether CD69 deficiency affected the inflammatory responses induced by BLM, we comprehensively investigated the differences in the expression of cytokines involving chemokines in the BALF. As shown in Figure 4A, the expression of several cytokines and chemokines in the BALF from WT and CD69-/- mice was induced by BLM. We focused on IL-6, MCP-1, TIMP-1, sTNF-R1, sTNF-R2, and MIP-1γ (Figure 4A &4B). These cytokines and chemokines were clearly induced by BLM in WT mice but less so in CD69-/- mice. Thus, the decreased expression of these factors may be closely related to the mechanisms underlying the attenuation of symptoms in CD69-/- mice, including the reduction in leukocytic infiltration and edema. An analysis by densitometer revealed that the expression levels of these cytokines and chemokines in BLM-treated CD69-/- mice were significantly lower than those in WT mice (Figure 4B).


Attenuation of lung inflammation and fibrosis in CD69-deficient mice after intratracheal bleomycin.

Yamauchi K, Kasuya Y, Kuroda F, Tanaka K, Tsuyusaki J, Ishizaki S, Matsunaga H, Iwamura C, Nakayama T, Tatsumi K - Respir. Res. (2011)

Effect of bleomycin treatment on cytokine expression in wild-type and CD69-deficient mice. (A) Cytokine array analyses of BALF 7 d after the instillation of BLM or PBS (sham treatment) in WT or CD69-/- mice. Cytokines with increased expression levels are boxed. (B) Expression levels of these cytokines in WT (black bars) and CD69-/- (white bars) mice. Each expression level was normalized by that of the positive control. The stimulation index is the ratio of the expression level of a cytokine in BLM-treated mice to that in sham-treated mice. Results are expressed as the mean (SEM) (n = 4 mice per group). *P < 0.05, **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198935&req=5

Figure 4: Effect of bleomycin treatment on cytokine expression in wild-type and CD69-deficient mice. (A) Cytokine array analyses of BALF 7 d after the instillation of BLM or PBS (sham treatment) in WT or CD69-/- mice. Cytokines with increased expression levels are boxed. (B) Expression levels of these cytokines in WT (black bars) and CD69-/- (white bars) mice. Each expression level was normalized by that of the positive control. The stimulation index is the ratio of the expression level of a cytokine in BLM-treated mice to that in sham-treated mice. Results are expressed as the mean (SEM) (n = 4 mice per group). *P < 0.05, **P < 0.01.
Mentions: To evaluate whether CD69 deficiency affected the inflammatory responses induced by BLM, we comprehensively investigated the differences in the expression of cytokines involving chemokines in the BALF. As shown in Figure 4A, the expression of several cytokines and chemokines in the BALF from WT and CD69-/- mice was induced by BLM. We focused on IL-6, MCP-1, TIMP-1, sTNF-R1, sTNF-R2, and MIP-1γ (Figure 4A &4B). These cytokines and chemokines were clearly induced by BLM in WT mice but less so in CD69-/- mice. Thus, the decreased expression of these factors may be closely related to the mechanisms underlying the attenuation of symptoms in CD69-/- mice, including the reduction in leukocytic infiltration and edema. An analysis by densitometer revealed that the expression levels of these cytokines and chemokines in BLM-treated CD69-/- mice were significantly lower than those in WT mice (Figure 4B).

Bottom Line: Cluster of differentiation 69 (CD69), an early activation marker antigen on T and B cells, is also expressed on activated macrophages and neutrophils, suggesting that CD69 may play a role in inflammatory diseases.Lung inflammation in the acute phase (7 dpi) was investigated by differential cell counts and cytokine array analyses of bronchoalveolar lavage fluid.In addition, lung fibrotic changes were evaluated at 14 dpi by histopathology and collagen assays.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Respirology, Graduate School of Medicine, Chiba University, Chiba, Japan.

ABSTRACT

Background: Cluster of differentiation 69 (CD69), an early activation marker antigen on T and B cells, is also expressed on activated macrophages and neutrophils, suggesting that CD69 may play a role in inflammatory diseases. To determine the effect of CD69 deficiency on bleomycin(BLM)-induced lung injury, we evaluated the inflammatory response following intratracheal BLM administration and the subsequent fibrotic changes in wild type (WT) and CD69-deficient (CD69-/-) mice.

Methods: The mice received a single dose of 3 mg/kg body weight of BLM and were sacrificed at 7 or 14 days post-instillation (dpi). Lung inflammation in the acute phase (7 dpi) was investigated by differential cell counts and cytokine array analyses of bronchoalveolar lavage fluid. In addition, lung fibrotic changes were evaluated at 14 dpi by histopathology and collagen assays. We also used reverse transcription polymerase chain reaction to measure the mRNA expression level of transforming growth factor β1 (TGF-β1) in the lungs of BLM-treated mice.

Results: CD69-/- mice exhibited less lung damage than WT mice, as shown by reductions in the following indices: (1) loss of body weight, (2) wet/dry ratio of lung, (3) cytokine levels in BALF, (4) histological evidence of lung injury, (5) lung collagen deposition, and (6) TGF-β1 mRNA expression in the lung.

Conclusion: The present study clearly demonstrates that CD69 plays an important role in the progression of lung injury induced by BLM.

Show MeSH
Related in: MedlinePlus