A CU scientist developed two mouse cell lines that expresses variants of the insulin B-chain residues linked to the mouse MHC class II molecules of beta and alpha I-AG7. These cell lines are derivatives of the mouse M12.C3 B cell line, which was originally derived from Balb/c mice that are negative for MHC class II molecules. The first cell line, called M12.C3-B:9-22(WT), expresses insulin B-chain residues 9-22 (SHLVEALYLVCGER) covalently linked to the beta chain of beta I-AG7. The second cell line, called M12.C3-B:9-22(RE), expresses the insulin B-chain residues 9-22 with mutations (SHLVERLYLVCGEE) that promote binding to I-AG7 in register 3. In order to express the above insulin residues, M12.C3 cells were transduced with a retroviral vector encoding the constructs. These cell lines can be used in drug screening, research, bioassays, etc. and as controls for binding of the anti-insulin peptide-MHC antibody: mAb287.
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