Chemiluminescence of non-differentiated THP-1 promocytes : developing an assay for screening anti-inflammatory milk proteins and peptides

Tompa, Gorazd (avtor), Laine, Aire (avtor), Pihlanto, Anne (avtor), Korhonen, Hannu (avtor), Rogelj, Irena (avtor), Marnila, Pertti (avtor)

Human leukemic THP-1 promonocytes are widely used as a model for peripheral blood monocytes. However, superoxide production during respiratory burst (RB) of non-differentiated THP-1 (nd-THP-1) cells is very low. Here we present a rapid and low-cost method for measuring the chemiluminescence (CL) of opsonized zymosan (OZ) induced RB which allows detection of Escherichia coli lipopolysaccharide (LPS) induced priming of nd-THP-1 cells on the basis of CL reaction kinetics. Maximum CL intensity obtained was 2.20+- 0.25 and 1.30+- 0.11 relative light units, while CL peak time was achieved at 18.1+- 2.6 and 28.7+- 1.3 min in primed and non-primed cells, respectively. The priming of nd-THP-1 cells with LPS evoked tzpical TNF-[alfa] and IL-6 production. We tested the effects of bovine lactoferrin and protein fractions from Lactobacillus helveticus BGRA43 fermented milk for potential anti-inflammatory effects on LPS primed nd-THP-1 cells. Four fractions were found to inhibit the OZ-induced CL in a dose-dependent manner (IC50 3-30 g/mL), while lactoferrin inhibited CL to a lesser extent (IC50 270? g/mL). These results suggest that measuring CL response of nd-THP-1 cells can serve as a method for screening anti-inflammatory compounds which could be used in reducing the risk of phagocyte-mediated inflammatory diseases.

• Jezik: Angleški jezik
• Ključne besede: mleko, beljakovine, peptidi, analizna kemija, kemiluminiscenca
• Vrsta gradiva: Delo ni kategorizirano
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Leto izida: 2011
• Št. strani: Str. 251-258
• Številčenje: Vol. 26, no. 4
• PID: 20.500.12556/RUL-36531
• UDK: 637.1:543
• ISSN pri članku: 1522-7235
• DOI: 10.1002/bio.1220
• COBISS.SI-ID: 2757768
• Datum objave v RUL: 10.07.2015

Gradivo je del revije

• Naslov: Luminescence
• Skrajšan naslov: Luminescence
• Založnik: Wiley & Sons
• ISSN: 1522-7235
• COBISS.SI-ID: 436108