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Titolo:
QUANTIFICATION OF C5A C5A(DESARG) IN BOVINE PLASMA, SERUM AND MILK/
Autore:
RAINARD P; SARRADIN P; PAAPE MJ; POUTREL B;
Indirizzi:
INRA,PATHOL INFECT & IMMUNOL LAB F-37380 NOUZILLY FRANCE ARS,IMMUNOL & DIS RESISTANCE LAB,INST LIVESTOCK & POULTRY SCI,USDA BELTSVILLE MD 20705
Titolo Testata:
Veterinary research
fascicolo: 1, volume: 29, anno: 1998,
pagine: 73 - 88
SICI:
0928-4249(1998)29:1<73:QOCCIB>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
COMPLEMENT ACTIVATION; MONOCLONAL-ANTIBODIES; STREPTOCOCCUS-AGALACTIAE; POLYACRYLAMIDE GELS; CHEMOTACTIC FACTORS; SIMPLIFIED ASSAY; HUMAN C5A; C3A; ANAPHYLATOXINS; RECEPTOR;
Keywords:
COMPLEMENT; C5A; C5; INFLAMMATION; ELISA; MASTITIS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
36
Recensione:
Indirizzi per estratti:
Citazione:
P. Rainard et al., "QUANTIFICATION OF C5A C5A(DESARG) IN BOVINE PLASMA, SERUM AND MILK/", Veterinary research, 29(1), 1998, pp. 73-88

Abstract

Complement activation generates two potent inflammatory mediators from C5, C5a and its derivative C5a(desArg), which results from the removal of the C-terminal arginine by ubiquitous carboxypeptidases. In thispaper we describe the purification of milligram amounts of bovine C5a(desArg) by a simplified procedure, and the preparation of mouse monoclonal antibodies (MAbs) to C5a/C5a(desArg) which do not recognize native C5.A MAb was used to develop a sandwich ELISA which made it possible to quantify levels of C5a/C5a(desArg) in bovine biological fluids. Small amounts (means +/- SEM) of C5a/C5a(desArg) were found in EDTA-plasma (0.58 +/- 0.06 ng.mL(-1)). The anticoagulant EDTA was more efficient than citrate or heparin in inhibiting in vitro activation of the complement system. Complement activation occurred during coagulation since the baseline concentration of C5a/C5a(desArg) (15.4 +/- 4.1 ng.mL(-1)) was higher than in plasma. Zymosan, a potent activator of the complement cascade, was used to generate C5a/C5a(desArg). The time-course of the reaction and the dose-effect of zymosan were investigated. Optimal conditions were incubation at 39 degrees C for 1 or 2 h with 2 mg of zymosan per mL of serum. The maximal concentration of C5a/C5a(desArg) attained in zymosan-activated serum was 4.28 +/- 0.14 mu g.mL(-1). Normal milk (from healthy, uninflamed mammary glands) contained on average 0.12 ng of C5a/C5a(desArg).mL(-1) (range 0.02-0.19 ng.mL(-1)). The maximal amount of C5a/C5a(desArg) which was generated in milk with zymosan was 1.1 ng.mL(-1) (range 0.68-2.17 ng.mL(-1)). In milk from quarters with subclinical infections by coagulase-negative staphylococci,values were 0.18 ng.mL(-1) and 2.37 ng.mL(-1) for spontaneous and zymosan-generated C5a/C5a(desArg) concentrations, respectively. In milk from Escherichia coli endotoxin-induced mastitis, C5a/C5a(desArg) concentrations (means of four cows) before and after zymosan activation reached 6.5 ng.mL(-1) and 55 ng.mL(-1), respectively. These results indicate that a C5-convertase can operate in normal milk, that only minute amounts of C5a/C5a(desArg) can be generated (less than 1/1 000 of plasma potential), but that much higher concentrations are reached in milkduring endotoxin-induced inflammation. The ELISA made it possible to determine normal ranges of C5a/C5a(desArg) in bovine blood plasma and in milk, and is a valuable tool to define the variations of its concentrations in exudates during inflammatory reactions. (C) Inra/Elsevier,Paris.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/01/21 alle ore 04:24:01