Details

Autor(en) / Beteiligte

• Percival-Alwyn, Jennifer L
• England, Elizabeth
• Kemp, Benjamin
• Rapley, Laura
• Davis, Nicola HE
• McCarthy, Grant R
• Majithiya, Jayesh B
• Corkill, Dominic J
• Welsted, Sarah
• Minton, Kevin
• Cohen, E Suzanne
• Robinson, Matthew J
• Dobson, Claire
• Wilkinson, Trevor CI
• Vaughan, Tristan J
• Groves, Maria AT
• Tigue, Natalie J

Titel

Generation of potent mouse monoclonal antibodies to self-proteins using T-cell epitope "tags"

Ist Teil von

• mAbs, 2015, Vol.7 (1), p.129-137

Ort / Verlag

Taylor & Francis

Erscheinungsjahr

2015

Link zum Volltext

Quelle

Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals

Beschreibungen/Notizen

• Immunization of mice or rats with a "non-self" protein is a commonly used method to obtain monoclonal antibodies, and relies on the immune system's ability to recognize the immunogen as foreign. Immunization of an antigen with 100% identity to the endogenous protein, however, will not elicit a robust immune response. To develop antibodies to mouse proteins, we focused on the potential for breaking such immune tolerance by genetically fusing two independent T-cell epitope-containing sequences (from tetanus toxin (TT) and diphtheria toxin fragment A (DTA)) to a mouse protein, mouse ST2 (mST2). Wild-type CD1 mice were immunized with three mST2 tagged proteins (Fc, TT and DTA) and the specific serum response was determined. Only in mice immunized with the T-cell epitope-containing antigens were specific mST2 serum responses detected; hybridomas generated from these mice secreted highly sequence-diverse IgGs that were capable of binding mST2 and inhibiting the interaction of mST2 with its ligand, mouse interleukin (IL)-33 (mIL-33). Of the hundreds of antibodies profiled, we identified five potent antibodies that were able to inhibit IL-33 induced IL-6 release in a mast cell assay; notably one such antibody was sufficiently potent to suppress IL-5 release and eosinophilia infiltration in an Alternaria alternata challenge mouse model of asthma. This study demonstrated, for the first time, that T-cell epitope-containing tags have the ability to break tolerance in wild-type mice to 100% conserved proteins, and it provides a compelling argument for the broader use of this approach to generate antibodies against any mouse protein or conserved ortholog.