developing an assay for screening anti-inflammatory milk proteins and peptides
Gorazd Tompa (Author), Aire Laine (Author), Anne Pihlanto (Author), Hannu Korhonen (Author), Irena Rogelj (Author), Pertti Marnila (Author)

Abstract

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;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UL BF - Biotechnical Faculty
UDC: 637.1:543
COBISS: 2757768 Link will open in a new window
ISSN: 1522-7235
Views: 1310
Downloads: 215
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Other data

Secondary language: Unknown
Type (COBISS): Not categorized
Pages: str. 251-258
Volume: ǂVol. ǂ26
Issue: ǂno. ǂ4
Chronology: 2011
DOI: 10.1002/bio.1220
ID: 1033738
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