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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

Monocyte subsets from CL patients produce MMP-9.PBMC were obtained from healthy subjects (HS) (n = 17), early CL (ECL) (n = 11) and CL patients (n = 28), and staining to CD14, CD16 and MMP-9 was performed. Frequency of monocyte subsets producing MMP-9 was determined ex-vivo by intracellular staining. Gate strategy to assess monocyte subsets based on CD14 and CD16 expression and frequency of MMP-9 in each subset of monocytes in HS (A) and CL patients (B). The gate strategy was done based on all minus one staining. C, Frequency of MMP-9-producing monocyte subsets from HS, ECL and CL patients. D, Mean fluorescence intensity (MFI) of MMP-9 positive cells in monocyte subsets from HS, ECL and CL patients. Data shown from a representative experiment of four performances. *p<0.05; **p<0.005; ***p<0.0005.
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pntd-0003282-g004: Monocyte subsets from CL patients produce MMP-9.PBMC were obtained from healthy subjects (HS) (n = 17), early CL (ECL) (n = 11) and CL patients (n = 28), and staining to CD14, CD16 and MMP-9 was performed. Frequency of monocyte subsets producing MMP-9 was determined ex-vivo by intracellular staining. Gate strategy to assess monocyte subsets based on CD14 and CD16 expression and frequency of MMP-9 in each subset of monocytes in HS (A) and CL patients (B). The gate strategy was done based on all minus one staining. C, Frequency of MMP-9-producing monocyte subsets from HS, ECL and CL patients. D, Mean fluorescence intensity (MFI) of MMP-9 positive cells in monocyte subsets from HS, ECL and CL patients. Data shown from a representative experiment of four performances. *p<0.05; **p<0.005; ***p<0.0005.

Mentions: Recently, three monocyte subsets have been described based on the expression of CD14 and CD16, subdividing them into classical, intermediate and non-classical monocytes [5]. In our study the gate strategy to access the monocyte subsets was defined based on CD14 and CD16 expression. To evaluate the contribution of monocyte subsets to MMP-9 production we performed intracellular ex-vivo staining of PBMC from patients with ECL, CL and HS which were then analyzed by flow cytometry. The frequency of MMP-9 in each subsets of monocyte was performed using an isotype control (Figure 4A and B). Our data shows that monocytes from patients with ECL and CL express more MMP-9 than monocytes from HS and that non-classical monocytes (CD14+CD16++) are the major source of MMP-9 in patients with CL (Figure 4C and 4D).


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)

Monocyte subsets from CL patients produce MMP-9.PBMC were obtained from healthy subjects (HS) (n = 17), early CL (ECL) (n = 11) and CL patients (n = 28), and staining to CD14, CD16 and MMP-9 was performed. Frequency of monocyte subsets producing MMP-9 was determined ex-vivo by intracellular staining. Gate strategy to assess monocyte subsets based on CD14 and CD16 expression and frequency of MMP-9 in each subset of monocytes in HS (A) and CL patients (B). The gate strategy was done based on all minus one staining. C, Frequency of MMP-9-producing monocyte subsets from HS, ECL and CL patients. D, Mean fluorescence intensity (MFI) of MMP-9 positive cells in monocyte subsets from HS, ECL and CL patients. Data shown from a representative experiment of four 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-g004: Monocyte subsets from CL patients produce MMP-9.PBMC were obtained from healthy subjects (HS) (n = 17), early CL (ECL) (n = 11) and CL patients (n = 28), and staining to CD14, CD16 and MMP-9 was performed. Frequency of monocyte subsets producing MMP-9 was determined ex-vivo by intracellular staining. Gate strategy to assess monocyte subsets based on CD14 and CD16 expression and frequency of MMP-9 in each subset of monocytes in HS (A) and CL patients (B). The gate strategy was done based on all minus one staining. C, Frequency of MMP-9-producing monocyte subsets from HS, ECL and CL patients. D, Mean fluorescence intensity (MFI) of MMP-9 positive cells in monocyte subsets from HS, ECL and CL patients. Data shown from a representative experiment of four performances. *p<0.05; **p<0.005; ***p<0.0005.
Mentions: Recently, three monocyte subsets have been described based on the expression of CD14 and CD16, subdividing them into classical, intermediate and non-classical monocytes [5]. In our study the gate strategy to access the monocyte subsets was defined based on CD14 and CD16 expression. To evaluate the contribution of monocyte subsets to MMP-9 production we performed intracellular ex-vivo staining of PBMC from patients with ECL, CL and HS which were then analyzed by flow cytometry. The frequency of MMP-9 in each subsets of monocyte was performed using an isotype control (Figure 4A and B). Our data shows that monocytes from patients with ECL and CL express more MMP-9 than monocytes from HS and that non-classical monocytes (CD14+CD16++) are the major source of MMP-9 in patients with CL (Figure 4C and 4D).

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