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Annexin‐A1 identified as the oral epithelial cell anti‐ Candida effector moiety
Author(s) -
Lilly E.A.,
Yano J.,
Fidel P.L.
Publication year - 2010
Publication title -
molecular oral microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 77
eISSN - 2041-1014
pISSN - 2041-1006
DOI - 10.1111/j.2041-1014.2010.00579.x
Subject(s) - annexin , porin , effector , annexin a2 , biology , western blot , blot , annexin a1 , protease , microbiology and biotechnology , cell , biochemistry , chemistry , enzyme , bacterial outer membrane , escherichia coli , gene
Summary Innate and adaptive immunity are considered critical to protection against mucosal candidal infections. Among innate anti‐ Candida mechanisms, oral and vaginal epithelial cells have antifungal activity. The mechanism is fungistatic, acid‐labile and includes a requirement for cell contact by intact, but not necessarily live, epithelial cells. The purpose of this study was to use the acid‐labile property to further characterize the effector moiety. Surface material extracted from phosphate‐buffered saline (PBS) ‐treated, but not acid‐treated, epithelial cells significantly inhibited the growth of Candida blastoconidia in a dose‐dependent manner which was abrogated by prior heat and protease treatment. Proteins extracted from PBS‐treated cells bound blastoconidia and hyphae more intensely than those from acid‐treated cells. Proteins from PBS‐treated cells eluted from Candida revealed two unique bands of approximately 33 and 45 kDa compared with acid‐treated cells. Mass spectrometry identified these proteins as Annexin‐A1 and actin, respectively. Oral epithelial cells stained positive for Annexin‐A1, but not actin. Western blots showed reduced Annexin‐A1 in proteins from acid‐treated epithelial cells compared with those from PBS‐treated epithelial cells. Lastly, it was demonstrated that immunoprecipitation of Annexin‐A1 from proteins extracted from PBS‐treated oral epithelial cells resulted in abrogation of inhibitory activity. Taken together, these results indicate that Annexin‐A1 is a strong candidate for the epithelial cell anti‐ Candida effector protein.