Naslov
The expression of galectin-3 on a monocyte/macropgahe-like THP-1 cell line

Autori
Novak, Ruđer ; Lepur, Adriana ; Dabelić, Sanja ; Dumić, Jerka

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
XXIV International Congress, Cytometry in the Age of Systems Biology / - Budimpešta : International Society for Analytical Cytology, 2008, 203-204

Skup
XXIV International Congress, Cytometry in the Age of Systems Biology

Mjesto i datum
Budimpešta, Mađarska, 17.05.2008. - 21.05.2008

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
galectin-3; monocyte/macrophages; flow cytometry

Sažetak
Galectin-3, a β -galactoside binding lectin exerts important roles in many biological processes (e.g. adhesion, proliferation, differentiation, apoptosis) thus being involved in numerous patophysiological processes (inflammation, neoplastic transformation, spreading metastases). It was also shown that galectin-3 is one of the key lectins in innate and acquired immunity. In general, galectin-3 is considered as a powerful pro-inflammatory signal, e.g. it triggers/promotes respiratory burst in monocytes and is a monocyte/macrophage chemoattractant, while targeted disruption of the galectin-3 gene in mice results in a reduced inflammatory response, namely attenuated phagocytic activity. It is speculated that intracellular galectin-3 contributes to the persistence of inflammation as an anti-apoptotic factor, promoting the survival of inflammatory cells. The aim of this study was to determine the level of membrane and intracellular expression of galectin-3 on a monocytic THP-1 cell line. The expression was determined by flow cytometry on a Beckmann Coulter FC500 cytometer, upon lipopolysaccharide (LPS) and phorbol 12-myristate-13-acetate (PMA) treatment. THP-1 cells were cultured in RPMI 1640 supplemented with 10% FCS and antibiotics. Upon treatment with 100 ng/ml E.coli LPS, samples were collected after 1, 3, 6, 12 and 24 hours. Following appropriate blocking, the cells were incubated with (rat) anti-human galectin-3 antibodies followed by FITC-labeled secondary (rabbit) anti-rat antibody. Dead cells were excluded as positive by 7AAD staining. The level of expression of membrane galectin-3 was also determined at 12, 24, 48 and 72 hours of PMA-induced differentiation. Macrophage-like morphology was confirmed by an elevated CD14 stain and adhesion of the cells to Petri-dishes. Surface galectin-3 expression was determined by flow cytometry using previously mentioned antibodies. THP-1 cells, activated by E. coli LPS, have markedly up-regulated expression of intracellular galectin-3 while the surface level of the galectin remains largely unchanged, in respect to control cells. The level of surface galectin-3 expression rises on the course of PMA-induced macrophage differntiation in respect to control cells. Given the importance of galectin-3 in different aspects of monocyte/macrophage physiology, further experiments are nesessary to elucidate the regulatory pathways of it’ s expression.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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