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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

CL patients have high levels of MMP-9 and low levels of TIMP-1 when compared with healthy subjects.PBMC were obtained from healthy subjects (HS) (n = 12), early CL (ECL) (n = 19) and CL patients (n = 18), and levels of MMP-9 (A) and TIMP-1 (B), determined by ELISA in culture supernatants of PBMCs upon stimulation with soluble Leishmania antigen (SLA) (5 µg/ml) for 72 hours. C, Ratio between MMP-9 and TIMP-1 in culture supernatants of PBMCs upon stimulation with SLA. The box-and-wickers plot shows the minimum, first quartile, median, third quartile, and maximum values of a set of data. The wickers represent error bars and those represent minimum and maximum values, indicating variability outside the upper and lower quartiles. Data shown from a representative experiment of five performances. *p<0.05; **p<0.005; ***p<0.0005.
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pntd-0003282-g002: CL patients have high levels of MMP-9 and low levels of TIMP-1 when compared with healthy subjects.PBMC were obtained from healthy subjects (HS) (n = 12), early CL (ECL) (n = 19) and CL patients (n = 18), and levels of MMP-9 (A) and TIMP-1 (B), determined by ELISA in culture supernatants of PBMCs upon stimulation with soluble Leishmania antigen (SLA) (5 µg/ml) for 72 hours. C, Ratio between MMP-9 and TIMP-1 in culture supernatants of PBMCs upon stimulation with SLA. The box-and-wickers plot shows the minimum, first quartile, median, third quartile, and maximum values of a set of data. The wickers represent error bars and those represent minimum and maximum values, indicating variability outside the upper and lower quartiles. Data shown from a representative experiment of five performances. *p<0.05; **p<0.005; ***p<0.0005.

Mentions: TIMPs are glycoproteins that inhibit MMPs and TIMP-1 specifically regulates the activity of MMP-9. Therefore, our next step was to determine the production MMP-9 and TIMP-1 by PBMC from HS and CL patients in the early and late phase of the disease. To do so, PBMC were stimulated in vitro with SLA or left untreated (media) for 72 hours and the production of MMP-9 and TIMP-1 were determined in the supernatants by ELISA. Our results show that CL patients produced high levels of MMP-9 and low levels of TIMP-1 when compared to HS (Figure 2A and 2B). Interestingly, PBMCs from CL patients produced high levels of MMP-9 even in the absence of stimulus, indicating that those cells are prone to produced MMP-9 when still in CL patient. A change in the ratio of MMPs and TIMPs may cause excessive degradation of the extracellular matrix and consequently tissue damage [28]. Therefore, we next sought to determine the ratio between MMP-9 production and TIMP-1. The high ratio MMP-9/TIMP-1 in cultures from CL individuals indicated an imbalance in the production of these enzymes in L. braziliensis infected individuals when compared to HS (Figure 2C).


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)

CL patients have high levels of MMP-9 and low levels of TIMP-1 when compared with healthy subjects.PBMC were obtained from healthy subjects (HS) (n = 12), early CL (ECL) (n = 19) and CL patients (n = 18), and levels of MMP-9 (A) and TIMP-1 (B), determined by ELISA in culture supernatants of PBMCs upon stimulation with soluble Leishmania antigen (SLA) (5 µg/ml) for 72 hours. C, Ratio between MMP-9 and TIMP-1 in culture supernatants of PBMCs upon stimulation with SLA. The box-and-wickers plot shows the minimum, first quartile, median, third quartile, and maximum values of a set of data. The wickers represent error bars and those represent minimum and maximum values, indicating variability outside the upper and lower quartiles. Data shown from a representative experiment of five performances. *p<0.05; **p<0.005; ***p<0.0005.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003282-g002: CL patients have high levels of MMP-9 and low levels of TIMP-1 when compared with healthy subjects.PBMC were obtained from healthy subjects (HS) (n = 12), early CL (ECL) (n = 19) and CL patients (n = 18), and levels of MMP-9 (A) and TIMP-1 (B), determined by ELISA in culture supernatants of PBMCs upon stimulation with soluble Leishmania antigen (SLA) (5 µg/ml) for 72 hours. C, Ratio between MMP-9 and TIMP-1 in culture supernatants of PBMCs upon stimulation with SLA. The box-and-wickers plot shows the minimum, first quartile, median, third quartile, and maximum values of a set of data. The wickers represent error bars and those represent minimum and maximum values, indicating variability outside the upper and lower quartiles. Data shown from a representative experiment of five performances. *p<0.05; **p<0.005; ***p<0.0005.
Mentions: TIMPs are glycoproteins that inhibit MMPs and TIMP-1 specifically regulates the activity of MMP-9. Therefore, our next step was to determine the production MMP-9 and TIMP-1 by PBMC from HS and CL patients in the early and late phase of the disease. To do so, PBMC were stimulated in vitro with SLA or left untreated (media) for 72 hours and the production of MMP-9 and TIMP-1 were determined in the supernatants by ELISA. Our results show that CL patients produced high levels of MMP-9 and low levels of TIMP-1 when compared to HS (Figure 2A and 2B). Interestingly, PBMCs from CL patients produced high levels of MMP-9 even in the absence of stimulus, indicating that those cells are prone to produced MMP-9 when still in CL patient. A change in the ratio of MMPs and TIMPs may cause excessive degradation of the extracellular matrix and consequently tissue damage [28]. Therefore, we next sought to determine the ratio between MMP-9 production and TIMP-1. The high ratio MMP-9/TIMP-1 in cultures from CL individuals indicated an imbalance in the production of these enzymes in L. braziliensis infected individuals when compared to HS (Figure 2C).

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