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Investigation of a new tumor-associated glycosylated antigen as target for dendritic cell vaccination in pancreatic cancer.

Béraud E, Collignon A, Franceschi C, Olive D, Lombardo D, Mas E - Oncoimmunology (2012)

Bottom Line: Glycoproteins, as valuable targets for dendritic cell (DC)-vaccination in cancers, remain an open question.Glycosylated structures, which are aberrantly modified during cancerisation, impact positively or negatively on glycoprotein immunogenicity.Here is presented an oncofetal glycovariant of bile-salt-dependent-lipase, expressed on human tumoral pancreas and efficiently processed by DC's, inducing T-lymphocyte activation.

View Article: PubMed Central - PubMed

Affiliation: INSERM; Marseille, France; Aix-Marseille Univ ; Centre de Recherche en Oncologie biologique et Oncopharmacologie; Marseille, France.

ABSTRACT
Glycoproteins, as valuable targets for dendritic cell (DC)-vaccination in cancers, remain an open question. Glycosylated structures, which are aberrantly modified during cancerisation, impact positively or negatively on glycoprotein immunogenicity. Here is presented an oncofetal glycovariant of bile-salt-dependent-lipase, expressed on human tumoral pancreas and efficiently processed by DC's, inducing T-lymphocyte activation.

No MeSH data available.


Related in: MedlinePlus

Figure 1. Uptake of J28 glycosylated antigen by iMoDC and intracellular localization. (A) Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) and Alexa 488-labeled ovalbumin (A488-OVA) uptakes were analyzed by confocal laser microscopy. After 5-d culture, iMoDC were loaded with A488-pBSDL-J28 (50 μg/mL) or A488-OVA (50 μg/mL) for 1h, washed, fixed, and counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies. (B) Uptake of Alexa 488 recombinant C-terminal glycopolypeptide-carrying J28 glycotope (A488-Cter-J28) by iMoDC. iMoDC were incubated for 1h at 37°C with A488-Cter-J28, washed, counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies, and analyzed by confocal microscopy. (C) Intracellular localization of A488-pBSDL-J28. iMoDC were incubated for 1h at 37°C with A488-pBSDL-J28 (50 µg/mL) and counterstained with antibodies directed against CD206, HLA-DR, HLA-DM, and CD107a. Colocalization of each intracellular marker (red) with A488-pBSDL-J28 (green) is indicated in yellow. Original magnification x630.
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Figure 1: Figure 1. Uptake of J28 glycosylated antigen by iMoDC and intracellular localization. (A) Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) and Alexa 488-labeled ovalbumin (A488-OVA) uptakes were analyzed by confocal laser microscopy. After 5-d culture, iMoDC were loaded with A488-pBSDL-J28 (50 μg/mL) or A488-OVA (50 μg/mL) for 1h, washed, fixed, and counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies. (B) Uptake of Alexa 488 recombinant C-terminal glycopolypeptide-carrying J28 glycotope (A488-Cter-J28) by iMoDC. iMoDC were incubated for 1h at 37°C with A488-Cter-J28, washed, counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies, and analyzed by confocal microscopy. (C) Intracellular localization of A488-pBSDL-J28. iMoDC were incubated for 1h at 37°C with A488-pBSDL-J28 (50 µg/mL) and counterstained with antibodies directed against CD206, HLA-DR, HLA-DM, and CD107a. Colocalization of each intracellular marker (red) with A488-pBSDL-J28 (green) is indicated in yellow. Original magnification x630.

Mentions: Figure 1 shows that human immature monocyte-derived DC’s (iMoDC) captures pBSDL-J28 as well as recombinant C-terminal polypeptide-J28. pBSDL-J28 binds to MR (CD206) expressed on iMoDC surface.12 Thus, oligosaccharides recognized by C-type lectin-like carbohydrate recognition domains of MR terminating in mannose, N-acetylglucosamine, and/or fucose residues, the latter being crucial elements of the J28 glycotope structure for mAbJ28 recognition, can be involved in DC’s binding (for a review see ref. 17). However, we cannot rule out that pBSDL-J28 binds to other receptors. Binding results in the internalization of pBSDL-J28 and its delivery into MHC class II compartment (MIIC) and late endosomes, where the Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) processing products were detected. A488-(glycosylated)-peptide epitopes co-localized in lysosomes with LAMP-1 (CD107a) and in late endosomes with HLA-DM. Thus, DC’s internalize tumoral antigens mainly by receptor mediated-endocytosis but also by macropinocytosis (not shown here) to direct them to MIIC.12


Investigation of a new tumor-associated glycosylated antigen as target for dendritic cell vaccination in pancreatic cancer.

Béraud E, Collignon A, Franceschi C, Olive D, Lombardo D, Mas E - Oncoimmunology (2012)

Figure 1. Uptake of J28 glycosylated antigen by iMoDC and intracellular localization. (A) Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) and Alexa 488-labeled ovalbumin (A488-OVA) uptakes were analyzed by confocal laser microscopy. After 5-d culture, iMoDC were loaded with A488-pBSDL-J28 (50 μg/mL) or A488-OVA (50 μg/mL) for 1h, washed, fixed, and counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies. (B) Uptake of Alexa 488 recombinant C-terminal glycopolypeptide-carrying J28 glycotope (A488-Cter-J28) by iMoDC. iMoDC were incubated for 1h at 37°C with A488-Cter-J28, washed, counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies, and analyzed by confocal microscopy. (C) Intracellular localization of A488-pBSDL-J28. iMoDC were incubated for 1h at 37°C with A488-pBSDL-J28 (50 µg/mL) and counterstained with antibodies directed against CD206, HLA-DR, HLA-DM, and CD107a. Colocalization of each intracellular marker (red) with A488-pBSDL-J28 (green) is indicated in yellow. Original magnification x630.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376954&req=5

Figure 1: Figure 1. Uptake of J28 glycosylated antigen by iMoDC and intracellular localization. (A) Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) and Alexa 488-labeled ovalbumin (A488-OVA) uptakes were analyzed by confocal laser microscopy. After 5-d culture, iMoDC were loaded with A488-pBSDL-J28 (50 μg/mL) or A488-OVA (50 μg/mL) for 1h, washed, fixed, and counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies. (B) Uptake of Alexa 488 recombinant C-terminal glycopolypeptide-carrying J28 glycotope (A488-Cter-J28) by iMoDC. iMoDC were incubated for 1h at 37°C with A488-Cter-J28, washed, counterstained with CD1a antibodies followed by mouse Alexa-594-secondary antibodies, and analyzed by confocal microscopy. (C) Intracellular localization of A488-pBSDL-J28. iMoDC were incubated for 1h at 37°C with A488-pBSDL-J28 (50 µg/mL) and counterstained with antibodies directed against CD206, HLA-DR, HLA-DM, and CD107a. Colocalization of each intracellular marker (red) with A488-pBSDL-J28 (green) is indicated in yellow. Original magnification x630.
Mentions: Figure 1 shows that human immature monocyte-derived DC’s (iMoDC) captures pBSDL-J28 as well as recombinant C-terminal polypeptide-J28. pBSDL-J28 binds to MR (CD206) expressed on iMoDC surface.12 Thus, oligosaccharides recognized by C-type lectin-like carbohydrate recognition domains of MR terminating in mannose, N-acetylglucosamine, and/or fucose residues, the latter being crucial elements of the J28 glycotope structure for mAbJ28 recognition, can be involved in DC’s binding (for a review see ref. 17). However, we cannot rule out that pBSDL-J28 binds to other receptors. Binding results in the internalization of pBSDL-J28 and its delivery into MHC class II compartment (MIIC) and late endosomes, where the Alexa 488-labeled pBSDL-J28 (A488-pBSDL-J28) processing products were detected. A488-(glycosylated)-peptide epitopes co-localized in lysosomes with LAMP-1 (CD107a) and in late endosomes with HLA-DM. Thus, DC’s internalize tumoral antigens mainly by receptor mediated-endocytosis but also by macropinocytosis (not shown here) to direct them to MIIC.12

Bottom Line: Glycoproteins, as valuable targets for dendritic cell (DC)-vaccination in cancers, remain an open question.Glycosylated structures, which are aberrantly modified during cancerisation, impact positively or negatively on glycoprotein immunogenicity.Here is presented an oncofetal glycovariant of bile-salt-dependent-lipase, expressed on human tumoral pancreas and efficiently processed by DC's, inducing T-lymphocyte activation.

View Article: PubMed Central - PubMed

Affiliation: INSERM; Marseille, France; Aix-Marseille Univ ; Centre de Recherche en Oncologie biologique et Oncopharmacologie; Marseille, France.

ABSTRACT
Glycoproteins, as valuable targets for dendritic cell (DC)-vaccination in cancers, remain an open question. Glycosylated structures, which are aberrantly modified during cancerisation, impact positively or negatively on glycoprotein immunogenicity. Here is presented an oncofetal glycovariant of bile-salt-dependent-lipase, expressed on human tumoral pancreas and efficiently processed by DC's, inducing T-lymphocyte activation.

No MeSH data available.


Related in: MedlinePlus