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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

Lesions of CL patients produce MMP-9.A, Heatmap showing expression of MMPs and TIMP-1 from microarray profile. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. Average fold change (FC) for each gene in lesion samples relative to normal skin controls is shown. B, Ratio between MMP-9 and TIMP-1 genes expression. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. C, MMP-9 levels in biopsies culture supernatants from CL patients (n = 6) and healthy subjects (HS) (n = 5), determined by ELISA after the biopsies been cultured for 12 h in absence of stimuli. **p<0.005.
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pntd-0003282-g001: Lesions of CL patients produce MMP-9.A, Heatmap showing expression of MMPs and TIMP-1 from microarray profile. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. Average fold change (FC) for each gene in lesion samples relative to normal skin controls is shown. B, Ratio between MMP-9 and TIMP-1 genes expression. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. C, MMP-9 levels in biopsies culture supernatants from CL patients (n = 6) and healthy subjects (HS) (n = 5), determined by ELISA after the biopsies been cultured for 12 h in absence of stimuli. **p<0.005.

Mentions: For whole genome expression microarray, lesion biopsies preserved in RNAlater (Qiagen) were homogenized using a rotor-stator and RNA was isolated using the RNeasy Plus kit (Qiagen). Biotin-labeled complementary RNA (cRNA) was generated using the Illumina TotalPrep RNA amplification kit (Ambion). RNA and cRNA quality were assessed on a BioAnalyzer (Agilent). Illumina HumanHT-12 version 4 expression beadchips were hybridized with cRNA from 26 L. braziliensis lesion biopsies and 10 biopses collected from uninfected donors. Arrays were scanned using a BeadStation 500GX and raw image files were processed using GenomeStudio v1.8 software (Illumina). Data was variance stabilized, robust-spline normalized, and quality control analysis carried out using the Lumi package [22] in Bioconductor/R. Differential expression analysis of the data using linear models and empirical Bayes methods [23] was carried out using the Limma package [24]. Data was deposited on the Gene Expression Omnibus (GEO) database for public access (GSE#GSE43880). Heat map tools available on GenePattern [25] were used to graphically display differentially regulated genes in Figure 1A.


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)

Lesions of CL patients produce MMP-9.A, Heatmap showing expression of MMPs and TIMP-1 from microarray profile. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. Average fold change (FC) for each gene in lesion samples relative to normal skin controls is shown. B, Ratio between MMP-9 and TIMP-1 genes expression. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. C, MMP-9 levels in biopsies culture supernatants from CL patients (n = 6) and healthy subjects (HS) (n = 5), determined by ELISA after the biopsies been cultured for 12 h in absence of stimuli. **p<0.005.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003282-g001: Lesions of CL patients produce MMP-9.A, Heatmap showing expression of MMPs and TIMP-1 from microarray profile. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. Average fold change (FC) for each gene in lesion samples relative to normal skin controls is shown. B, Ratio between MMP-9 and TIMP-1 genes expression. Biopsies from CL patients (n = 26) and normal skin (n = 10) were obtained and unbiased microarray was performed on biopsies mRNA. C, MMP-9 levels in biopsies culture supernatants from CL patients (n = 6) and healthy subjects (HS) (n = 5), determined by ELISA after the biopsies been cultured for 12 h in absence of stimuli. **p<0.005.
Mentions: For whole genome expression microarray, lesion biopsies preserved in RNAlater (Qiagen) were homogenized using a rotor-stator and RNA was isolated using the RNeasy Plus kit (Qiagen). Biotin-labeled complementary RNA (cRNA) was generated using the Illumina TotalPrep RNA amplification kit (Ambion). RNA and cRNA quality were assessed on a BioAnalyzer (Agilent). Illumina HumanHT-12 version 4 expression beadchips were hybridized with cRNA from 26 L. braziliensis lesion biopsies and 10 biopses collected from uninfected donors. Arrays were scanned using a BeadStation 500GX and raw image files were processed using GenomeStudio v1.8 software (Illumina). Data was variance stabilized, robust-spline normalized, and quality control analysis carried out using the Lumi package [22] in Bioconductor/R. Differential expression analysis of the data using linear models and empirical Bayes methods [23] was carried out using the Limma package [24]. Data was deposited on the Gene Expression Omnibus (GEO) database for public access (GSE#GSE43880). Heat map tools available on GenePattern [25] were used to graphically display differentially regulated genes in Figure 1A.

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