State of differentiation defines buccal epithelial cell affinity for cross-linking to Candida albicans Hwp1

Candida albicans utilizes mammalian cell-associated transglutaminase (TGase) activity to adhere covalently to human buccal epithelial cells (BECs) through Hyphal Wall Protein 1. Little is known about the factors leading to the identity and appearance of Hwp1 binding partners on cells lining the oral cavity. The observation that BECs vary in their ability to attach to C. albicans germ tubes and to bind recombinant Hwp1 (rHwp1) suggested that differentiation may play a role in affinity for germ tube attachment. Individual BECs were characterized for differentiation status and rHwp1 binding. rHwp1 bound to the more terminally differentiated cells displaying SPR3 and keratin 13 but not to less differentiated cells with abundant involucrin. Sequential expression of involucrin followed by SPR3 in oral keratinocytes was demonstrated using stratified organotypic cultures and a feeder layer system with the OKF6/TERT-2 cell line. Increased cross-linking of the lysine analogue 5-(biotinamido) pentylamine to cultured OKF6/TERT-2 cell proteins accompanied this increased expression of SPR3. Western blot analysis demonstrated the presence of rHwp1 cross-links to proteins from BECs or from OKF6/TERT-2 cells that had been mechanically dislodged from culture dishes. Therefore, the differentiation of SPR3 positive from involucrin positive cells is correlated with the acquisition of affinity for cross-linking to rHwp1 and covalent adhesion of germ tubes to BECs.