Advanced

Galectin-3 endocytosis by carbohydrate independent and dependent pathways in different macrophage like cell types.

Lepur, Adriana LU ; Carlsson, Michael LU ; Novak, Ruđer; Dumić, Jerka; Nilsson, Ulf and Leffler, Hakon (2012) In Biochimica et Biophysica Acta 1820(7). p.804-818
Abstract
BACKGROUND: Galectin-3 (the Mac-2 antigen) is abundantly expressed in both macrophage like cells and certain non-macrophage cells. We have studied endocytosis of galectin-3 as one important step relevant for its function, and compared it between variants of a macrophage like cell line, and non-macrophage cells.



METHODS: Endocytosis of galectin-3 was observed by fluorescence microscopy and measured by flow cytometry. The endocytosis mechanism was analysed using galectin-3 mutants, galectin-3 inhibitors and endocytic pathways inhibitors in the human leukaemia THP-1 cell line differentiated into naïve (M0), classical (M1) or alternatively activated (M2) macrophage like cells, and the non-macrophage cell lines HFL-1... (More)
BACKGROUND: Galectin-3 (the Mac-2 antigen) is abundantly expressed in both macrophage like cells and certain non-macrophage cells. We have studied endocytosis of galectin-3 as one important step relevant for its function, and compared it between variants of a macrophage like cell line, and non-macrophage cells.



METHODS: Endocytosis of galectin-3 was observed by fluorescence microscopy and measured by flow cytometry. The endocytosis mechanism was analysed using galectin-3 mutants, galectin-3 inhibitors and endocytic pathways inhibitors in the human leukaemia THP-1 cell line differentiated into naïve (M0), classical (M1) or alternatively activated (M2) macrophage like cells, and the non-macrophage cell lines HFL-1 fibroblasts and SKBR3 breast carcinoma.



RESULTS: Galectin-3 endocytosis in non-macrophage cells and M2 cells was blocked by lactose and a potent galectin-3 inhibitor TD139, and also by the R186S mutation in the galectin-3 carbohydrate recognition domain (CRD). In M1 cells galectin-3 endocytosis could be inhibited only by chlorpromazine and by interference with the non-CRD N-terminal part of galectin-3. In all the cell types galectin-3 entered early endosomes within 5-10min, to be subsequently targeted mainly to non-degradative vesicles, where it remained even after 24h.



CONCLUSIONS: Galectin-3 endocytosis in M1 cells is receptor mediated and carbohydrate independent, while in M2 cells it is CRD mediated, although the non-CRD galectin-3 domain is also involved. General significance The demonstration that galectin-3 endocytosis in M1 macrophages is carbohydrate independent and different from M2 macrophages and non-macrophage cells, suggests novel, immunologically significant interactions between phagocytic cells, galectin-3 and its ligands. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochimica et Biophysica Acta
volume
1820
issue
7
pages
804 - 818
publisher
Elsevier
external identifiers
  • WOS:000305366100003
  • PMID:22450157
  • Scopus:84860675027
ISSN
0006-3002
DOI
10.1016/j.bbagen.2012.02.018
language
English
LU publication?
yes
id
f63aac35-6350-4bf7-866a-e224429f2246 (old id 2431447)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/22450157?dopt=Abstract
date added to LUP
2012-04-03 13:51:14
date last changed
2017-01-15 04:28:36
@article{f63aac35-6350-4bf7-866a-e224429f2246,
  abstract     = {BACKGROUND: Galectin-3 (the Mac-2 antigen) is abundantly expressed in both macrophage like cells and certain non-macrophage cells. We have studied endocytosis of galectin-3 as one important step relevant for its function, and compared it between variants of a macrophage like cell line, and non-macrophage cells. <br/><br>
<br/><br>
METHODS: Endocytosis of galectin-3 was observed by fluorescence microscopy and measured by flow cytometry. The endocytosis mechanism was analysed using galectin-3 mutants, galectin-3 inhibitors and endocytic pathways inhibitors in the human leukaemia THP-1 cell line differentiated into naïve (M0), classical (M1) or alternatively activated (M2) macrophage like cells, and the non-macrophage cell lines HFL-1 fibroblasts and SKBR3 breast carcinoma. <br/><br>
<br/><br>
RESULTS: Galectin-3 endocytosis in non-macrophage cells and M2 cells was blocked by lactose and a potent galectin-3 inhibitor TD139, and also by the R186S mutation in the galectin-3 carbohydrate recognition domain (CRD). In M1 cells galectin-3 endocytosis could be inhibited only by chlorpromazine and by interference with the non-CRD N-terminal part of galectin-3. In all the cell types galectin-3 entered early endosomes within 5-10min, to be subsequently targeted mainly to non-degradative vesicles, where it remained even after 24h. <br/><br>
<br/><br>
CONCLUSIONS: Galectin-3 endocytosis in M1 cells is receptor mediated and carbohydrate independent, while in M2 cells it is CRD mediated, although the non-CRD galectin-3 domain is also involved. General significance The demonstration that galectin-3 endocytosis in M1 macrophages is carbohydrate independent and different from M2 macrophages and non-macrophage cells, suggests novel, immunologically significant interactions between phagocytic cells, galectin-3 and its ligands.},
  author       = {Lepur, Adriana and Carlsson, Michael and Novak, Ruđer and Dumić, Jerka and Nilsson, Ulf and Leffler, Hakon},
  issn         = {0006-3002},
  language     = {eng},
  number       = {7},
  pages        = {804--818},
  publisher    = {Elsevier},
  series       = {Biochimica et Biophysica Acta},
  title        = {Galectin-3 endocytosis by carbohydrate independent and dependent pathways in different macrophage like cell types.},
  url          = {http://dx.doi.org/10.1016/j.bbagen.2012.02.018},
  volume       = {1820},
  year         = {2012},
}