Biosensor incorporating cell barrier architectures for detecting Staphylococcus aureus alpha toxin

被引:10
作者
Ghosh, Gargi
Bachas, Leonidas G.
Anderson, Kimberly W. [1 ]
机构
[1] Univ Kentucky, Dept Chem & Mat Engn, Lexington, KY 40506 USA
[2] Univ Kentucky, Dept Chem, Lexington, KY 40506 USA
基金
美国国家航空航天局;
关键词
potassium-selective electrodes; biosensor; alpha toxin; intercellular gaps; silver staining;
D O I
10.1007/s00216-006-0949-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Alpha toxin is a common virulent factor of Staphylococcus aureus and is believed to play crucial roles in pathogenicity induced by S. aureus. Alpha toxin is also known to induce permeability to endothelial cell monolayers in vitro due to the formation of interendothelial gaps. The present study is directed towards measuring alpha toxin using a whole-cell-based biosensor. The biosensor, consisting of a confluent monolayer of human umbilical vein endothelial cells (HUVECs) on a potassium ion-selective electrode, takes advantage of cell permeability dysfunction to detect the presence of small quantities of alpha toxin. When a confluent monolayer of cells was formed on the membrane surface, the response of the electrode toward the marker ion, potassium, was inhibited. Upon exposing this sensor to varying concentrations of alpha toxin for 20 min, an increase in sensor response to potassium was observed. The response thus obtained was indirectly related to the concentration of alpha toxin. The detection limit of this sensor for alpha toxin was found to be 0.1 ng/ml. Cell monolayers were stained with silver nitrate to quantify the formation of intercellular gaps as well as to study the effect of this toxin on HUVECs morphology. A strong positive correlation was observed between the response obtained from the biosensor and the area of the intercellular gaps. Silver staining also revealed the tendency of cells to round up upon being exposed to alpha toxin.
引用
收藏
页码:567 / 574
页数:8
相关论文
共 48 条
  • [1] Endothelial gaps: Time course of formation and closure in inflamed venules of rats
    Baluk, P
    Hirata, A
    Thurston, G
    Fujiwara, T
    Neal, CR
    Michel, CC
    McDonald, DM
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 1997, 272 (01) : L155 - L170
  • [2] HUMAN HYPERIMMUNE GLOBULIN PROTECTS AGAINST THE CYTO-TOXIC ACTION OF STAPHYLOCOCCAL ALPHA-TOXIN INVITRO AND INVIVO
    BHAKDI, S
    MANNHARDT, U
    MUHLY, M
    HUGO, F
    RONNEBERGER, H
    HUNGERER, KD
    [J]. INFECTION AND IMMUNITY, 1989, 57 (10) : 3214 - 3220
  • [3] ALPHA-TOXIN OF STAPHYLOCOCCUS-AUREUS
    BHAKDI, S
    TRANUMJENSEN, J
    [J]. MICROBIOLOGICAL REVIEWS, 1991, 55 (04) : 733 - 751
  • [4] STAPHYLOCOCCAL ALPHA-TOXIN PROMOTES BLOOD-COAGULATION VIA ATTACK ON HUMAN-PLATELETS
    BHAKDI, S
    MUHLY, M
    MANNHARDT, U
    HUGO, F
    KLAPETTEK, K
    MUELLERECKHARDT, C
    ROKA, L
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1988, 168 (02) : 527 - 542
  • [5] Adrenomedullin reduces Staphylococcus aureus α-toxin-induced rat ileum microcirculatory damage
    Brell, B
    Temmesfeld-Wollbrück, B
    Altzschner, I
    Frisch, E
    Schmeck, B
    Hocke, AC
    Suttorp, N
    Hippenstiel, S
    [J]. CRITICAL CARE MEDICINE, 2005, 33 (04) : 819 - 826
  • [6] Effect of surface-attached heparin on the response of potassium-selective electrodes
    Brooks, HA
    Allen, JR
    Feldhoff, PW
    Bachas, LG
    [J]. ANALYTICAL CHEMISTRY, 1996, 68 (08) : 1439 - 1443
  • [7] RECOMMENDATIONS FOR NOMENCLATURE OF ION-SELECTIVE ELECTRODES - (IUPAC RECOMMENDATIONS 1994)
    BUCK, RP
    LINDNER, E
    [J]. PURE AND APPLIED CHEMISTRY, 1994, 66 (12) : 2527 - 2536
  • [8] Burns AR, 1997, J IMMUNOL, V159, P2893
  • [9] Alpha-toxin is required for biofilm formation by Staphylococcus aureus
    Caiazza, NC
    O'Toole, GA
    [J]. JOURNAL OF BACTERIOLOGY, 2003, 185 (10) : 3214 - 3217
  • [10] POTENTIOMETRIC ION-SELECTIVE AND BIO-SELECTIVE ELECTRODES BASED ON ASYMMETRIC CELLULOSE-ACETATE MEMBRANES
    CHA, GS
    MEYERHOFF, ME
    [J]. TALANTA, 1989, 36 (1-2) : 271 - 278