Immunization of mice with human DC derived-exosomes and generation of hybridomas that produce antibodies recognizing at least the human exosomes

Data obtained from a proteomic analysis of human exosomes performed by Sebastian AMIGORENA from the DC-THERA consortium allowed us to suspect a number of proteins as a potential candidate target for our hybridoma 13.

We have immunized mice with human DC derived-exosomes and generated hybridomas that produce antibodies recognizing at least the human exosomes. Out of 20 hybridomas, only one appears to recognize specifically DC exosomes and not tumor exosomes nor autologous monocytes or DC or PBL. The Ab produced by this N°13 hybridoma is binding to a protein of about 30kDa. The proteomic analysis of human exosomes allowed us to suspect these proteins as a potential candidate target for our hybridoma 13:

- Signalling: 14-3-3 beta (28 kDa), 14-3-3 eta (28 kDa), 14-3-3 gamma (28 kDa), 14-3-3 zeta (28 kDa)
- Adhesion: CD9 (25 kDa), galectin 3 (27 kDa)
- Enzymes: glucose 6 phosphate isomerase (32 kDa)
- H2-M: MB2d (28 kDa), MB2k1 (28 kDa), MB2p (28 kDa), MB2q1 (28 kDa), MB2r (28 kDa), MB2w3 (28 kDa), MB2w5 (28 kDa)
- Lipid rafts: prohibitin (29 kDa), stomatin (31 kDa)
- Membrane fusion: syntaxin 7 (29 kDa)
- Cytoskeleton: syntenin (32 kDa)
- Others: ribosomal protein L6 (32 kDa)

A mass spectrometry after 2D electrophoresis will be performed on exosomal lysates to formally identify the candidate protein recognized by our hybridoma 13. We hope to obtain a neutralizing Ab to investigate the role of exosome in immunity in vivo.

• organism type
• Mus musculus,
• Homo sapiens

• molecule type
• PHB,
• galectin,
• glucose 6 phosphate isomerase,
• ribosomal protein L6

created over 16 years ago (2 March 2009)    last modified over 13 years ago (28 September 2011)   [ RDF ]   [ RelFinder ]