[Screening of ADAMs that can mediate the generation of soluble MHC I]

To screen ADAMs that can mediate the generation of soluble MHCI using HEK293 cells are study model.

ADAM8, ADAM10, ADAM15 or ADAM17 cDNA expressing vector s were separately constructed with pcDNA3. 1/V5 as vectors and were transfected into HEK293 cells. Hygromycin B was used to screen the cells that stably expressed ADAMs. After HEK293 cells were sell-surface labeled with biotin and cultured for another 4 h and soluble MHC I the was examined by SDS-PAGE, Western blot and enhanced chemiluminescence.

HEK293 cells stably expressing ADAMs were successfully obtained and the over-expression of ADAM10 or ADAM17 increased the release of soluble MHC I.

ADAM10 and ADAM17 have the potentials of mediating the generation of soluble MHC I.