Document Detail

Increase in milk metalloproteinase activity and vascular permeability in bovine endotoxin-induced and naturally occurring Escherichia coli mastitis.
MedLine Citation:
PMID:  11943315     Owner:  NLM     Status:  MEDLINE    
An endotoxin-induced mastitis model was used to study permeability changes associated with increased milk matrix metalloproteinase (MMP) activity in early inflammation. One quarter of two cows was inoculated with endotoxin (Escherichia coli 055:B5). Blood, milk, and whey were collected before and repeatedly after inoculation for 48 h. The profile and amounts of gelatinolytic MMPs were determined by zymography; gelatinase A (72 kD MMP-2) and gelatinase B (92 kD MMP-9) were identified by Western immunoblotting. Bovine serum albumin (BSA) and trypsin inhibitor capacity (TIC) were used as markers of capillary permeability with parallel examination of neutrophil penetration from blood to milk. Five clinical E. coli mastitis milk samples and five milk samples from cows with healthy udders were analyzed to detect whether increased levels of gelatinolytic MMP-2 and MMP-9 have a role in naturally occurring mastitis with endotoxin involvement. Milk MMP levels increased 2h after the endotoxin challenge. Both MMP-2 and MMP-9 were involved in this early proteolytic event. These increased MMP levels are associated with increased capillary permeability, evidenced first by the penetration of small molecular weight proteins, such as BSA and TIC. Later, 6-12h post endotoxin inoculation, neutrophilic leucocytes also entered the site, as they require larger tissue damage in basal membrane and interstitial tissue structures than BSA and TIC to extravasate. In naturally occurring disease, increased MMP-2 and MMP-9 levels were detected in milk. Thus, gelatinases, especially MMP-2, are involved in the early degradation of the blood-milk barrier, which precedes the penetration of blood-derived cellular components into milk in endotoxin-induced mastitis. In the future, measuring MMP in milk/whey might be a useful tool for diagnosing early mastitis.
S M Raulo; T Sorsa; T Tervahartiala; T Latvanen; E Pirilä; J Hirvonen; P Maisi
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Veterinary immunology and immunopathology     Volume:  85     ISSN:  0165-2427     ISO Abbreviation:  Vet. Immunol. Immunopathol.     Publication Date:  2002 Mar 
Date Detail:
Created Date:  2002-04-10     Completed Date:  2002-06-12     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8002006     Medline TA:  Vet Immunol Immunopathol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  137-45     Citation Subset:  IM    
Faculty of Veterinary Medicine, Department of Clinical Veterinary Sciences, Helsinki University, P.O. Box 57, Hämeentie 57, Helsinki FIN-00014, Finland.
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MeSH Terms
Blotting, Western / veterinary
Capillary Permeability / immunology
Electrophoresis, Polyacrylamide Gel / veterinary
Endotoxins / metabolism,  toxicity
Escherichia coli* / metabolism
Escherichia coli Infections / enzymology,  immunology,  veterinary*
Leukocyte Count / veterinary
Mastitis, Bovine / blood,  enzymology*,  immunology
Matrix Metalloproteinase 2 / biosynthesis*,  metabolism
Matrix Metalloproteinase 9 / biosynthesis*,  metabolism
Milk / enzymology*
Milk Proteins / analysis
Plant Proteins / blood
Serum Albumin, Bovine / analysis
Trypsin Inhibitors
alpha-Amylases / antagonists & inhibitors
Reg. No./Substance:
0/Endotoxins; 0/Milk Proteins; 0/Plant Proteins; 0/Serum Albumin, Bovine; 0/Trypsin Inhibitors; 0/whey protein; EC; EC Metalloproteinase 2; EC Metalloproteinase 9

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