Limits...
The Stimulation of CD147 Induces MMP-9 Expression through ERK and NF-kappaB in Macrophages: Implication for Atherosclerosis.

Kim JY, Kim WJ, Kim H, Suk K, Lee WH - Immune Netw (2009)

Bottom Line: Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas.These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: The School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT

Background: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.

Methods: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface.

Results: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-kappaB. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of IkappaB, and nuclear translocation of NF-kappaB p65 and p50 subunits.

Conclusion: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

No MeSH data available.


Related in: MedlinePlus

The stimulation of CD147 induced the expression of MMP-9 in THP-1 cells. (A) THP-1 cells were stimulated with anti-CD147 mAb or isotype-matching mouse IgG (mIgG) that had been immobilized at indicated concentrations. LPS was used as a positive control. Some of the cells were also stimulated with antibodies that had been heat inactivated at 95℃ for 2 hr. Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (B) THP-1 cells were stimulated as in (A) and culture supernatants were concentrated (×10) and subjected to Western blot analysis using MMP-9 specific antibody. As loading control, the picture of the membrane used for Western blot analysis that had been stained with Coomassie Brilliant Blue is shown at the lower panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2803300&req=5

Figure 3: The stimulation of CD147 induced the expression of MMP-9 in THP-1 cells. (A) THP-1 cells were stimulated with anti-CD147 mAb or isotype-matching mouse IgG (mIgG) that had been immobilized at indicated concentrations. LPS was used as a positive control. Some of the cells were also stimulated with antibodies that had been heat inactivated at 95℃ for 2 hr. Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (B) THP-1 cells were stimulated as in (A) and culture supernatants were concentrated (×10) and subjected to Western blot analysis using MMP-9 specific antibody. As loading control, the picture of the membrane used for Western blot analysis that had been stained with Coomassie Brilliant Blue is shown at the lower panel.

Mentions: Since macrophages in atherosclerotic plaques express CD147, monocyte/macrophage cell lines were used to test whether they express CD147. As shown in Fig. 2A, both THP-1 and U937 cells expressed high levels of CD147. Stimulation of THP-1 cells with CypA did not affect the expression levels of CD147, probably because the basal expression level of CD147 was already high (data not shown). The expression of CD147 in THP-1 was also confirmed using RT-PCR (Fig. 2B). THP-1 cells were then used to study the signaling pathway initiated from CD147. Since stimulation of THP-1 cells with CypA induced the expression of MMP-9 (20), CD147 on the surface of THP-1 cells were stimulated with anti-CD147 mAb and the cellular responses were analyzed. Anti-CD147 mAb was used instead of CypA, to exclude the possibility that CypA may stimulate other yet unknown cellular receptors. Stimulation of the cells with immobilized anti-CD147 mAb induced the secretion of MMP-9 (Fig. 3A). The expression levels of MMP-2, which is known to be unaffected by cellular activation status, are shown as the internal control. Isotype-matching mouse IgG failed to induce the expression of MMP-9 indicating that the induction of MMP-9 requires specific interaction between CD147 and the antibody. Furthermore, heat inactivation of anti-CD147 mAb abolished the effect indicating that the activation was not induced by endotoxins that are heat resistant. The induction of MMP-9 expression was also confirmed in the protein level using Western blot analysis (Fig. 3B).


The Stimulation of CD147 Induces MMP-9 Expression through ERK and NF-kappaB in Macrophages: Implication for Atherosclerosis.

Kim JY, Kim WJ, Kim H, Suk K, Lee WH - Immune Netw (2009)

The stimulation of CD147 induced the expression of MMP-9 in THP-1 cells. (A) THP-1 cells were stimulated with anti-CD147 mAb or isotype-matching mouse IgG (mIgG) that had been immobilized at indicated concentrations. LPS was used as a positive control. Some of the cells were also stimulated with antibodies that had been heat inactivated at 95℃ for 2 hr. Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (B) THP-1 cells were stimulated as in (A) and culture supernatants were concentrated (×10) and subjected to Western blot analysis using MMP-9 specific antibody. As loading control, the picture of the membrane used for Western blot analysis that had been stained with Coomassie Brilliant Blue is shown at the lower panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2803300&req=5

Figure 3: The stimulation of CD147 induced the expression of MMP-9 in THP-1 cells. (A) THP-1 cells were stimulated with anti-CD147 mAb or isotype-matching mouse IgG (mIgG) that had been immobilized at indicated concentrations. LPS was used as a positive control. Some of the cells were also stimulated with antibodies that had been heat inactivated at 95℃ for 2 hr. Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (B) THP-1 cells were stimulated as in (A) and culture supernatants were concentrated (×10) and subjected to Western blot analysis using MMP-9 specific antibody. As loading control, the picture of the membrane used for Western blot analysis that had been stained with Coomassie Brilliant Blue is shown at the lower panel.
Mentions: Since macrophages in atherosclerotic plaques express CD147, monocyte/macrophage cell lines were used to test whether they express CD147. As shown in Fig. 2A, both THP-1 and U937 cells expressed high levels of CD147. Stimulation of THP-1 cells with CypA did not affect the expression levels of CD147, probably because the basal expression level of CD147 was already high (data not shown). The expression of CD147 in THP-1 was also confirmed using RT-PCR (Fig. 2B). THP-1 cells were then used to study the signaling pathway initiated from CD147. Since stimulation of THP-1 cells with CypA induced the expression of MMP-9 (20), CD147 on the surface of THP-1 cells were stimulated with anti-CD147 mAb and the cellular responses were analyzed. Anti-CD147 mAb was used instead of CypA, to exclude the possibility that CypA may stimulate other yet unknown cellular receptors. Stimulation of the cells with immobilized anti-CD147 mAb induced the secretion of MMP-9 (Fig. 3A). The expression levels of MMP-2, which is known to be unaffected by cellular activation status, are shown as the internal control. Isotype-matching mouse IgG failed to induce the expression of MMP-9 indicating that the induction of MMP-9 requires specific interaction between CD147 and the antibody. Furthermore, heat inactivation of anti-CD147 mAb abolished the effect indicating that the activation was not induced by endotoxins that are heat resistant. The induction of MMP-9 expression was also confirmed in the protein level using Western blot analysis (Fig. 3B).

Bottom Line: Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas.These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: The School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT

Background: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.

Methods: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface.

Results: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-kappaB. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of IkappaB, and nuclear translocation of NF-kappaB p65 and p50 subunits.

Conclusion: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

No MeSH data available.


Related in: MedlinePlus