Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages

Abstract

The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins.

Involvement of thapsigargin– and cyclopiazonic acid–sensitive pumps in the rescue of TMEM165‐associated glycosylation defects by Mn2+

“…N-glycans in 5% aqueous acetic acid were lyophylized overnight. They were permethylated and spotted onto a matrix-assisted desorption plate and analyzed by matrix-assisted desorption ionization-time of flight mass spectrometry on a 4800 Proteomics Analyzer Mass Spectrometer (Applied Biosystems; Thermo Fisher Scientific), as described by Delannoy et al (13). Each spectrum resulted from the accumulation of 10,000 spectra and shows glycans structures from m/z 1500 up to 3000.…”

Autofluorescence mediated red spherulocyte sorting provides insights into the source of spinochromes in sea urchins

“…The results from a recent study indicate that lectin-based FACS could serve as a flow cytometry based strategy to purify distinct purple sea urchin (S. purpuratus) coelomocytes 25 . Lectins are known to target glycosylation patterns presented by individual cell types [26][27][28][29] and this interaction could thus be utilised to define different immune cell populations. For instance, peanut agglutinin (PNA), a plant lectin, facilitates the identification of activated germinal centre B cells 30 .…”

OGT Controls the Expression and the Glycosylation of E‐cadherin, and Affects Glycosphingolipid Structures in Human Colon Cell Lines

“…The mid mode (mass range m / z 1000–3000) was used with laser power set at 85 for two shots in 100 locations per spot. Assignment was made tentatively from calculated mass composition and a literature search …”