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Matrix metalloproteinase 9 production by monocytes is enhanced by TNF and participates in the pathology of human cutaneous Leishmaniasis.

Campos TM, Passos ST, Novais FO, Beiting DP, Costa RS, Queiroz A, Mosser D, Scott P, Carvalho EM, Carvalho LP - PLoS Negl Trop Dis (2014)

Bottom Line: Culture was performed during 72 hours, stimulating the cells with SLA, levels of MMP-9 and TIMP-1 in the supernatants were determined by ELISA.We observed that cells from CL lesions secrete high amounts of MMP-9 when compared to healthy subjects.We also observed that TNF produced in high level during CL contributes to MMP-9 production.

View Article: PubMed Central - PubMed

Affiliation: Serviço de Imunologia, Universidade Federal da Bahia, Salvador, Bahia, Brazil.

ABSTRACT

Introduction: Cutaneous leishmaniasis (CL) due to L.braziliensis infection is characterized by a strong inflammatory response with high levels of TNF and ulcer development. Less attention has been given to the role of mononuclear phagocytes to this process. Monocytes constitute a heterogeneous population subdivided into classical, intermediate and non-classical, and are known to migrate to inflammatory sites and secrete inflammatory mediators. TNF participates in the induction of matrix metalloproteinases (MMPs). MMP-9 is an enzyme that degrades basal membrane and its activity is controlled by the tissue inhibitor of metalloproteinase.

Methods: Mononuclear cells were obtained from ex-vivo labeling sub-populations of monocytes and MMP-9, and the frequency was determined by flow cytometry. Culture was performed during 72 hours, stimulating the cells with SLA, levels of MMP-9 and TIMP-1 in the supernatants were determined by ELISA.

Results: We observed that cells from CL lesions secrete high amounts of MMP-9 when compared to healthy subjects. Although MMP-9 was produced by monocytes, non-classical ones were the main source of this enzyme. We also observed that TNF produced in high level during CL contributes to MMP-9 production.

Conclusions: These observations emphasize the role of monocytes, TNF and MMP-9 in the pathogenesis of L. braziliensis infection.

No MeSH data available.


Related in: MedlinePlus

TNF enhances MMP-9 production in CL patients.A, PBMC from healthy subjects (HS) (n = 12) and CL patients (n = 19) were cultured in the presence or absence of SLA (5 µg/ml) for 72 hours, and TNF concentrations were determined by ELISA on supernatants. B, PBMC from HS (n = 5) were cultured in the presence or absence of recombinant TNF (5 ng/ml) for 24 hours and the levels of MMP-9 were determined by ELISA. C, PBMC from HS (n = 6) and CL patients (n = 6) was cultured in presence or absence of anti-TNF antibodies (10 µg/ml) for 24 hours. The levels of MMP-9 were determined by ELISA on supernatants. *p<0.05; ***p<0.0005.
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pntd-0003282-g005: TNF enhances MMP-9 production in CL patients.A, PBMC from healthy subjects (HS) (n = 12) and CL patients (n = 19) were cultured in the presence or absence of SLA (5 µg/ml) for 72 hours, and TNF concentrations were determined by ELISA on supernatants. B, PBMC from HS (n = 5) were cultured in the presence or absence of recombinant TNF (5 ng/ml) for 24 hours and the levels of MMP-9 were determined by ELISA. C, PBMC from HS (n = 6) and CL patients (n = 6) was cultured in presence or absence of anti-TNF antibodies (10 µg/ml) for 24 hours. The levels of MMP-9 were determined by ELISA on supernatants. *p<0.05; ***p<0.0005.

Mentions: TNF is an inflammatory cytokine that contributes to tissue damage by different mechanisms, including by induction of MMP-9 expression [31]. To determine if TNF production contributes to MMP-9 production in CL patients, we first measured TNF levels in supernatants of SLA-stimulated PBMC from HS and CL patients. As previously demonstrated [32], CL patients produced high levels of TNF in response to SLA (Figure 5A). In order to determine if the TNF produced had the ability to increase MMP-9 expression, we added recombinant TNF to PBMC cultures from HS and determined MMP-9 expression in the supernatants by ELISA. Our results show that there was a 5-fold increase in MMP-9 production in unstimulated PBMC cultures from healthy individuals upon exogenous addition of recombinant TNF (Figure 5B). Finally, we asked if TNF was playing a role in MMP-9 production in CL. In order to address this question, we added anti-TNF antibodies to PBMC cultures from HS and CL patients (Figure 5C) and we found that the blockage of TNF inhibits the production of MMP-9 in CL patients. Altogether these data show that TNF, in part, regulates MMP-9 production in CL patients.


Matrix metalloproteinase 9 production by monocytes is enhanced by TNF and participates in the pathology of human cutaneous Leishmaniasis.

Campos TM, Passos ST, Novais FO, Beiting DP, Costa RS, Queiroz A, Mosser D, Scott P, Carvalho EM, Carvalho LP - PLoS Negl Trop Dis (2014)

TNF enhances MMP-9 production in CL patients.A, PBMC from healthy subjects (HS) (n = 12) and CL patients (n = 19) were cultured in the presence or absence of SLA (5 µg/ml) for 72 hours, and TNF concentrations were determined by ELISA on supernatants. B, PBMC from HS (n = 5) were cultured in the presence or absence of recombinant TNF (5 ng/ml) for 24 hours and the levels of MMP-9 were determined by ELISA. C, PBMC from HS (n = 6) and CL patients (n = 6) was cultured in presence or absence of anti-TNF antibodies (10 µg/ml) for 24 hours. The levels of MMP-9 were determined by ELISA on supernatants. *p<0.05; ***p<0.0005.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230914&req=5

pntd-0003282-g005: TNF enhances MMP-9 production in CL patients.A, PBMC from healthy subjects (HS) (n = 12) and CL patients (n = 19) were cultured in the presence or absence of SLA (5 µg/ml) for 72 hours, and TNF concentrations were determined by ELISA on supernatants. B, PBMC from HS (n = 5) were cultured in the presence or absence of recombinant TNF (5 ng/ml) for 24 hours and the levels of MMP-9 were determined by ELISA. C, PBMC from HS (n = 6) and CL patients (n = 6) was cultured in presence or absence of anti-TNF antibodies (10 µg/ml) for 24 hours. The levels of MMP-9 were determined by ELISA on supernatants. *p<0.05; ***p<0.0005.
Mentions: TNF is an inflammatory cytokine that contributes to tissue damage by different mechanisms, including by induction of MMP-9 expression [31]. To determine if TNF production contributes to MMP-9 production in CL patients, we first measured TNF levels in supernatants of SLA-stimulated PBMC from HS and CL patients. As previously demonstrated [32], CL patients produced high levels of TNF in response to SLA (Figure 5A). In order to determine if the TNF produced had the ability to increase MMP-9 expression, we added recombinant TNF to PBMC cultures from HS and determined MMP-9 expression in the supernatants by ELISA. Our results show that there was a 5-fold increase in MMP-9 production in unstimulated PBMC cultures from healthy individuals upon exogenous addition of recombinant TNF (Figure 5B). Finally, we asked if TNF was playing a role in MMP-9 production in CL. In order to address this question, we added anti-TNF antibodies to PBMC cultures from HS and CL patients (Figure 5C) and we found that the blockage of TNF inhibits the production of MMP-9 in CL patients. Altogether these data show that TNF, in part, regulates MMP-9 production in CL patients.

Bottom Line: Culture was performed during 72 hours, stimulating the cells with SLA, levels of MMP-9 and TIMP-1 in the supernatants were determined by ELISA.We observed that cells from CL lesions secrete high amounts of MMP-9 when compared to healthy subjects.We also observed that TNF produced in high level during CL contributes to MMP-9 production.

View Article: PubMed Central - PubMed

Affiliation: Serviço de Imunologia, Universidade Federal da Bahia, Salvador, Bahia, Brazil.

ABSTRACT

Introduction: Cutaneous leishmaniasis (CL) due to L.braziliensis infection is characterized by a strong inflammatory response with high levels of TNF and ulcer development. Less attention has been given to the role of mononuclear phagocytes to this process. Monocytes constitute a heterogeneous population subdivided into classical, intermediate and non-classical, and are known to migrate to inflammatory sites and secrete inflammatory mediators. TNF participates in the induction of matrix metalloproteinases (MMPs). MMP-9 is an enzyme that degrades basal membrane and its activity is controlled by the tissue inhibitor of metalloproteinase.

Methods: Mononuclear cells were obtained from ex-vivo labeling sub-populations of monocytes and MMP-9, and the frequency was determined by flow cytometry. Culture was performed during 72 hours, stimulating the cells with SLA, levels of MMP-9 and TIMP-1 in the supernatants were determined by ELISA.

Results: We observed that cells from CL lesions secrete high amounts of MMP-9 when compared to healthy subjects. Although MMP-9 was produced by monocytes, non-classical ones were the main source of this enzyme. We also observed that TNF produced in high level during CL contributes to MMP-9 production.

Conclusions: These observations emphasize the role of monocytes, TNF and MMP-9 in the pathogenesis of L. braziliensis infection.

No MeSH data available.


Related in: MedlinePlus