De-aluminated zeolite Y as a tool to study endocytosis, a delivery system revealing differences between human peripheral dendritic cells

被引:7
作者
Andersson, L. I. M. [1 ]
Eriksson, H. [1 ]
机构
[1] Malmo Univ, Fac Hlth & Soc, Dept Biomed Lab Sci, S-20506 Malmo, Sweden
关键词
D O I
10.1111/j.1365-3083.2007.01948.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
We report the use of nanometre-sized zeolite particles as a novel approach to follow the endosomal acidification and proteolysis inside a viable cell. The method was verified by using human peripheral monocytes, a well known endocytosing cell population. Zeolite particles were subsequently used to investigate the endocytosing mechanisms of human peripheral dendritic cells (DC). Probes detecting pH neutral and acidic endosomes were adsorbed to de-aluminated zeolite Y, and used to detect endocytosis in immature human peripheral blood DC. Both the myeloid (mDC) and the plasmacytoid (pDC) dendritic cell subsets had an endocytosing capacity comparable with peripheral blood monocytes. However, the majority of both subsets of DC retained their endosomes at a neutral pH during the first hours after endocytosis and only a small number of the mDC showed any formation of acidic endosomes. Proteolytic degradation of endocytosed proteins was detected using self-quenched DQ-ovalbumin adsorbed to zeolite particles. Interestingly, a clear difference in proteolytic degradation of endocytosed ovalbumin was observed between the two subsets of DC. The mDC showed an efficient degradation of ovalbumin, while the pDC population displayed no or only minor proteolytic degradation. In conclusion, zeolite particles provide a useful tool to study the endocytosing mechanisms, and an efficient carrier of bio-molecules into the endosomal pathways of viable cells.
引用
收藏
页码:52 / 61
页数:10
相关论文
共 22 条
[1]   Recent advances in dendritic cell biology (vol 25, pg 87, 2005) [J].
Adams, S ;
O'Neill, DW ;
Bhardwaj, N .
JOURNAL OF CLINICAL IMMUNOLOGY, 2005, 25 (03) :175-+
[2]   Mechanisms of phagocytosis [J].
Allen, LAH ;
Aderem, A .
CURRENT OPINION IN IMMUNOLOGY, 1996, 8 (01) :36-40
[3]   Dendritic cells and the control of immunity [J].
Banchereau, J ;
Steinman, RM .
NATURE, 1998, 392 (6673) :245-252
[4]   Processing and presentation of antigens by dendritic cells: implications for vaccines [J].
Bhardwaj, N .
TRENDS IN MOLECULAR MEDICINE, 2001, 7 (09) :388-394
[5]   REMOVAL OF TRITON X-100 AND SDS FROM PROTEIN SOLUTIONS WITH ZEOLITE-Y [J].
BLUM, Z ;
ERIKSSON, H .
ACTA CHEMICA SCANDINAVICA, 1990, 44 (05) :531-532
[6]  
Breck D.W, 1974, ZEOLITE MOL SIEVES
[7]   Ultra-stable zeolites -: a tool for in-cell chemistry [J].
Dahm, Å ;
Eriksson, H .
JOURNAL OF BIOTECHNOLOGY, 2004, 111 (03) :279-290
[8]   TEMPORAL CHANGES OF LYSOSOME AND PHAGOSOME PH DURING PHAGOLYSOSOME FORMATION IN MACROPHAGES - STUDIES BY FLUORESCENCE SPECTROSCOPY [J].
GEISOW, MJ ;
HART, PD ;
YOUNG, MR .
JOURNAL OF CELL BIOLOGY, 1981, 89 (03) :645-652
[9]   The endocytic pathway: A mosaic of domains [J].
Gruenberg, J .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2001, 2 (10) :721-730
[10]   Conditions for the adsorption of proteins on ultrastable zeolite Y and its use in protein purification [J].
Klint, D ;
Eriksson, H .
PROTEIN EXPRESSION AND PURIFICATION, 1997, 10 (02) :247-255