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)

URLURL - Presentation file, Visit http://onlinelibrary.wiley.com/doi/10.1002/bio.1220/abstract This link opens in a new window

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.

Keywords:mleko, beljakovine, peptidi, analizna kemija, kemiluminiscenca
Work type:Not categorized (r6)
Tipology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Number of pages:str. 251-258
Numbering:Vol. 26, no. 4
ISSN on article:1522-7235
DOI:10.1002/bio.1220 Link is opened in a new window
COBISS.SI-ID:2757768 Link is opened in a new window
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Record is a part of a journal

Shortened title:Luminescence
Publisher:Wiley & Sons
COBISS.SI-ID:436108 This link opens in a new window

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