title: Effects of immunomodulatory and organism-associated molecules on the permeability of an in vitro blood-brain barrier model to amphotericin B and fluconazole.
abstract: Amphotericin B (AMB) is used to treat fungal infections of the central nervous system (CNS). However, AMB shows poor penetration into the CNS and little is known about the factors affecting its permeation through the blood-brain barrier (BBB). Therefore, we studied immunomodulatory and organism-associated molecules affecting the permeability of an in vitro BBB model to AMB. We examined the effects of interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), lipopolysaccharide (LPS), lipoteichoic acid (LTA), zymosan (ZYM), dexamethasone (DEX), cyclosporine, and tacrolimus on transendothelial electrical resistance (TEER); endothelial tight junctions; filamentous actin; and permeability to deoxycholate AMB (DAMB), liposomal AMB (LAMB), and fluconazole. Proinflammatory cytokines and organism-associated molecules significantly decreased the mean TEER by 40.7 to 100% (P < or = 0.004). DEX increased the mean TEER by 18.2 to 26.4% (P < or = 0.04). TNF-alpha and LPS increased the permeability to AMB by 8.2 to 14.5% compared to that for the controls (1.1 to 2.4%) (P < or = 0.04). None of the other molecules affected the model's permeability to AMB. By comparison, the BBB model's permeability to fluconazole was >78% under all conditions studied, without significant differences between the controls and the experimental groups. LPS and TNF-alpha decreased tight-junction protein zona occludens 1 (ZO-1) between endothelial cells. In conclusion, IL-1beta, ZYM, and LTA increased the permeability of the BBB to small ions but not to AMB, whereas TNF-alpha and LPS, which disrupted the endothelial layer integrity, increased the permeability to AMB.
authors.affiliation: Immunocompromised Host Section, Pediatric Oncology Branch, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
authors.forename: Vasilios
authors.initials: V
authors.lastname: Pyrgos
authors.forename: Diane
authors.initials: D
authors.lastname: Mickiene
authors.forename: Tin
authors.initials: T
authors.lastname: Sein
authors.forename: Margaret
authors.initials: M
authors.lastname: Cotton
authors.forename: Andrea
authors.initials: A
authors.lastname: Fransesconi
authors.forename: Isaac
authors.initials: I
authors.lastname: Mizrahi
authors.forename: Martha
authors.initials: M
authors.lastname: Donoghue
authors.forename: Nikkida
authors.initials: N
authors.lastname: Bundrant
authors.forename: Su-Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Matthew
authors.initials: M
authors.lastname: Hardwick
authors.forename: Shmuel
authors.initials: S
authors.lastname: Shoham
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Walsh
mesh_terms: D000666:Amphotericin B / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D000935:Antifungal Agents / Q000378:metabolism* / Q000494:pharmacology; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology*; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D015725:Fluconazole / Q000378:metabolism* / Q000494:pharmacology; D007155:Immunologic Factors / Q000494:pharmacology*; D051381:Rats; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D016207:Cytokines; D007155:Immunologic Factors; D000666:Amphotericin B; D015725:Fluconazole