CD38 appears to be a promising candidate in antibody therapy; it is upregulated on cell surfaces in many lymphoid tumors and undergoes rapid internalization after interaction with antibodies. The receptor-mediated endocytosis allows conjugating toxins/drugs that promote suicide only of the malignant cells. Here, we describe the preparation of CD38-immunoliposomes and test their functionality by incubating them with CD38+/- cells. Liposomes were prepared by extrusion of a lipid mixture containing a biotinylated polyethylene glycol-phospholipid and loaded with 5(6)-carboxyfluorescein. The anti-CD38 antibody (IB4) was biotinylated and then linked to streptavidin molecules; streptavidin acts like a bridge between the antibody and the biotinylated lipid of the liposomes. CD38+/- cells were incubated either with liposomes or immunoliposomes and analyzed by fluorescence microscopy and cytofluorimetry. The results indicated a specific mechanism of internalization, owing to CD38-mediated endocytosis, where CD38+ cells incubated with immunoliposomes scored top fluorescence levels. This coupling strategy, based on the use of a streptavidin bridge to prepare immunoliposomes, does not interfere with the cellular functionality and its broad potential use represents a great advantage. Here IB4, a murine monoclonal anti-CD38 antibody, was used to simplify the experiments, but the coupling procedure may be suitable also with human antibodies, against CD38 or other human markers.

Orciani, M., Cavaletti, G., Fino, V., Mattioli Belmonte, M., Tredici, G., Bruni, P., et al. (2008). Exploiting CD38-mediated endocytosis for immunoliposome internalization. ANTI-CANCER DRUGS, 19(6), 599-605 [10.1097/CAD.0b013e3282ffd673].

Exploiting CD38-mediated endocytosis for immunoliposome internalization

CAVALETTI, GUIDO ANGELO;TREDICI, GIOVANNI;
2008

Abstract

CD38 appears to be a promising candidate in antibody therapy; it is upregulated on cell surfaces in many lymphoid tumors and undergoes rapid internalization after interaction with antibodies. The receptor-mediated endocytosis allows conjugating toxins/drugs that promote suicide only of the malignant cells. Here, we describe the preparation of CD38-immunoliposomes and test their functionality by incubating them with CD38+/- cells. Liposomes were prepared by extrusion of a lipid mixture containing a biotinylated polyethylene glycol-phospholipid and loaded with 5(6)-carboxyfluorescein. The anti-CD38 antibody (IB4) was biotinylated and then linked to streptavidin molecules; streptavidin acts like a bridge between the antibody and the biotinylated lipid of the liposomes. CD38+/- cells were incubated either with liposomes or immunoliposomes and analyzed by fluorescence microscopy and cytofluorimetry. The results indicated a specific mechanism of internalization, owing to CD38-mediated endocytosis, where CD38+ cells incubated with immunoliposomes scored top fluorescence levels. This coupling strategy, based on the use of a streptavidin bridge to prepare immunoliposomes, does not interfere with the cellular functionality and its broad potential use represents a great advantage. Here IB4, a murine monoclonal anti-CD38 antibody, was used to simplify the experiments, but the coupling procedure may be suitable also with human antibodies, against CD38 or other human markers.
Articolo in rivista - Articolo scientifico
Membrane Glycoproteins; K562 Cells; Endocytosis; Liposomes; Antigens, CD38; Antibodies, Monoclonal; Tetradecanoylphorbol Acetate; Humans; Biotinylation
English
lug-2008
19
6
599
605
none
Orciani, M., Cavaletti, G., Fino, V., Mattioli Belmonte, M., Tredici, G., Bruni, P., et al. (2008). Exploiting CD38-mediated endocytosis for immunoliposome internalization. ANTI-CANCER DRUGS, 19(6), 599-605 [10.1097/CAD.0b013e3282ffd673].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/20349
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