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

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

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.

• Language: English
• Keywords: mleko, beljakovine, peptidi, analizna kemija, kemiluminiscenca
• Work type: Not categorized
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty
• Year: 2011
• Number of pages: Str. 251-258
• Numbering: Vol. 26, no. 4
• PID: 20.500.12556/RUL-36531
• UDC: 637.1:543
• ISSN on article: 1522-7235
• DOI: 10.1002/bio.1220
• COBISS.SI-ID: 2757768
• Publication date in RUL: 10.07.2015

Record is a part of a journal

• Title: Luminescence
• Shortened title: Luminescence
• Publisher: Wiley & Sons
• ISSN: 1522-7235
• COBISS.SI-ID: 436108