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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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Related in: MedlinePlus

Expression of CD69 on macrophages in the lung. The lung from WT mice at 2 dpi with PBS (sham treatment) (A) or BLM (B) were subjected to immunohistochemical staining with an anti-CD69 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively. The control lung from WT mice was subjected to immunohistochemical staining with an anti-gp36 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively (C). Each arrowhead points to an interstitial macrophage. Each asterisk indicates an alveolar macrophage. All sections were co-stained with DAPI. Each bar represents 50 μm.
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Figure 7: Expression of CD69 on macrophages in the lung. The lung from WT mice at 2 dpi with PBS (sham treatment) (A) or BLM (B) were subjected to immunohistochemical staining with an anti-CD69 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively. The control lung from WT mice was subjected to immunohistochemical staining with an anti-gp36 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively (C). Each arrowhead points to an interstitial macrophage. Each asterisk indicates an alveolar macrophage. All sections were co-stained with DAPI. Each bar represents 50 μm.

Mentions: As shown in Figure 7A, Iba1-positive macrophages were observed in the lungs from WT PBS-injected mice but rarely exhibited CD69-like immunoreactivity. On the other hand, Iba1+/CD69+ macrophages were clearly observed in the lung from WT BLM-treated mice at 2 dpi (Figure 7B), indicating that the expression of CD69 was induced in the macrophages exposed to BLM. In addition, Iba1 recognized alveolar and interstitial macrophages, suggesting that BLM induced CD69 expression in the two types of macrophages (Figure 7C). At 2 dpi after BLM injection, T cells and neutrophils were barely detected as infiltrating cells in the lung (data not shown).


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)

Expression of CD69 on macrophages in the lung. The lung from WT mice at 2 dpi with PBS (sham treatment) (A) or BLM (B) were subjected to immunohistochemical staining with an anti-CD69 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively. The control lung from WT mice was subjected to immunohistochemical staining with an anti-gp36 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively (C). Each arrowhead points to an interstitial macrophage. Each asterisk indicates an alveolar macrophage. All sections were co-stained with DAPI. Each bar represents 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 7: Expression of CD69 on macrophages in the lung. The lung from WT mice at 2 dpi with PBS (sham treatment) (A) or BLM (B) were subjected to immunohistochemical staining with an anti-CD69 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively. The control lung from WT mice was subjected to immunohistochemical staining with an anti-gp36 antibody and an anti-Iba1 antibody, followed by a reaction with Alexa Fluor 594-conjugated and Alexa Fluor 488-conjugated secondary antibodies, respectively (C). Each arrowhead points to an interstitial macrophage. Each asterisk indicates an alveolar macrophage. All sections were co-stained with DAPI. Each bar represents 50 μm.
Mentions: As shown in Figure 7A, Iba1-positive macrophages were observed in the lungs from WT PBS-injected mice but rarely exhibited CD69-like immunoreactivity. On the other hand, Iba1+/CD69+ macrophages were clearly observed in the lung from WT BLM-treated mice at 2 dpi (Figure 7B), indicating that the expression of CD69 was induced in the macrophages exposed to BLM. In addition, Iba1 recognized alveolar and interstitial macrophages, suggesting that BLM induced CD69 expression in the two types of macrophages (Figure 7C). At 2 dpi after BLM injection, T cells and neutrophils were barely detected as infiltrating cells in the lung (data not shown).

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