Substance P-induced cadherin expression and its signal transduction in a cloned human corneal epithelial cell line

Araki-Sasaki, Kaoru; Aizawa, Shin; Hara, Masaki; Nakamura, Masatsugu; Iwase, Osamu; Nakata, Katsuhiko; Sasaki, Yutaka; Mano, Tomoo; Handa, Hiroshi; Tano, Yasuo

doi:10.1002/(sici)1097-4652(200002)182:2<189::aid-jcp7>3.0.co;2-9

Cited by 70 publications

(35 citation statements)

References 23 publications

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“…1 ìg cholera toxin ml À1 ; vitamins; 40 % Hanks F12; 40 % Dulbecco's modified Eagle's medium; 0 . 5 % DMSO; 10 ng epidermal growth factor ml À1 ) (Invitrogen) at 37 8C in a 5 % CO 2 incubator as previously described (Araki-Sasaki et al, 2000). Acanthamoeba isolates (5 3 10 5 amoebae per well) were incubated with cell monolayers in serum-free medium (RPMI 1640 containing 2 mM glutamine, 1 mM pyruvate and non-essential amino acids) at 37 8C in a 5 % CO 2 incubator for up to 24 h. At the end of this incubation period, supernatants were collected and cytotoxicity was determined by measuring lactate dehydrogenase release using a cytotoxicity detection kit (Roche Applied Science) as previously described (Khan & Tareen, 2003).…”

Section: Methodsmentioning

confidence: 99%

Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

Maghsood

Sissons

Rezaian

et al. 2005

Journal of Medical Microbiology

129

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The majority of the keratitis-causing Acanthamoeba isolates are genotype T4. In an attempt to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence of this genotype, Acanthamoeba genotypes were determined for 13 keratitis isolates and 12 environmental isolates from Iran. Among 13 clinical isolates, eight (61 . 5 %) belonged to T4, two (15 . 3 %) belonged to T3 and three (23 %) belonged to the T2 genotype. In contrast, the majority of 12 environmental isolates tested in the present study belonged to T2 (7/12, 58 . 3 %), followed by 4/12 T4 isolates (33 . 3 %). In addition, the genotypes of six new Acanthamoeba isolates from UK keratitis cases were determined. Of these, five (83 . 3 %) belonged to T4 and one was T3 (16 . 6 %), supporting the expected high frequency of T4 in Acanthamoeba keratitis. In total, the genotypes of 24 Acanthamoeba keratitis isolates from the UK and Iran were determined. Of these, 17 belonged to T4 (70 . 8 %), three belonged to T2 (12 . 5 %), three belonged to T3 (12 . 5 %) and one belonged to T11 (4 . 1 %), confirming that T4 is the predominant genotype (÷ 2 ¼ 4 . 167; P ¼ 0 . 0412) in Acanthamoeba keratitis.

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Section: Methodsmentioning

confidence: 99%

Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

Maghsood

Sissons

Rezaian

et al. 2005

Journal of Medical Microbiology

129

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“…Immortalized corneal epithelial cells (16) were used to evaluate epithelial coverage. Collagen, collagen-TERP, and collagen-TERP5 hydrogels (500 m thick for easy handling) were embedded separately on top of a collagenbased matrix that consisted of a mixture of blended neutralized type I rat-tail tendon collagen (0.3%, wt͞vol; Becton Dickinson) and chondroitin 6-sulfate (1:5, wt͞wt), crosslinked with 0.02% (vol͞vol) glutaraldehyde (followed by glycine termination of unreacted aldehyde groups) and then thermogelled at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Cellular and nerve regeneration within a biosynthetic extracellular matrix for corneal transplantation

Li¹,

Carlsson²,

Lohmann³

et al. 2004

American Journal of Ophthalmology

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Our objective was to determine whether key properties of extracellular matrix (ECM) macromolecules can be replicated within tissue-engineered biosynthetic matrices to influence cellular properties and behavior. To achieve this, hydrated collagen and Nisopropylacrylamide copolymer-based ECMs were fabricated and tested on a corneal model. The structural and immunological simplicity of the cornea and importance of its extensive innervation for optimal functioning makes it an ideal test model. In addition, corneal failure is a clinically significant problem. Matrices were therefore designed to have the optical clarity and the proper dimensions, curvature, and biomechanical properties for use as corneal tissue replacements in transplantation. In vitro studies demonstrated that grafting of the laminin adhesion pentapeptide motif, YIGSR, to the hydrogels promoted epithelial stratification and neurite in-growth. Implants into pigs' corneas demonstrated successful in vivo regeneration of host corneal epithelium, stroma, and nerves. In particular, functional nerves were observed to rapidly regenerate in implants. By comparison, nerve regeneration in allograft controls was too slow to be observed during the experimental period, consistent with the behavior of human cornea transplants. Other corneal substitutes have been produced and tested, but here we report an implantable matrix that performs as a physiologically functional tissue substitute and not simply as a prosthetic device. These biosynthetic ECM replacements should have applicability to many areas of tissue engineering and regenerative medicine, especially where nerve function is required.regenerative medicine ͉ tissue engineering ͉ cornea ͉ implantation ͉ innervation

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“…Briefly, HCEC were grown in RPMI 1640 containing 10 % fetal calf serum and 2 mM glutamine. Under these conditions, HCEC exhibited corneal epithelial cell-specific properties, as described previously (Araki-Sasaki et al, 2000). For adhesion and cytotoxicity assays, both HBMEC and HCEC were grown in 24-well plates by incubating 10 6 cells per well.…”

Section: Human Brain Microvascular Endothelial Cells (Hbmec)mentioning

confidence: 99%

“…Immortalized HCEC were routinely cultured as described previously (Araki-Sasaki et al, 2000). Briefly, HCEC were grown in RPMI 1640 containing 10 % fetal calf serum and 2 mM glutamine.…”

Section: Human Brain Microvascular Endothelial Cells (Hbmec)mentioning

confidence: 99%

Acanthamoeba induces cell-cycle arrest in host cells

Sissons

Alsam

Jayasekera

et al. 2004

Journal of Medical Microbiology

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Acanthamoeba can cause fatal granulomatous amoebic encephalitis (GAE) and eye keratitis. However, the pathogenesis and pathophysiology of these emerging diseases remain unclear. In this study, the effects of Acanthamoeba on the host cell cycle using human brain microvascular endothelial cells (HBMEC) and human corneal epithelial cells (HCEC) were determined. Two isolates of Acanthamoeba belonging to the T1 genotype (GAE isolate) and T4 genotype (keratitis isolate) were used, which showed severe cytotoxicity on HBMEC and HCEC, respectively. No tissue specificity was observed in their ability to exhibit binding to the host cells. To determine the effects of Acanthamoeba on the host cell cycle, a cell-cycle-specific gene array was used. This screened for 96 genes specific for host cell-cycle regulation. It was observed that Acanthamoeba inhibited expression of genes encoding cyclins F and G1 and cyclin-dependent kinase 6, which are proteins important for cell-cycle progression. Moreover, upregulation was observed of the expression of genes such as GADD45A and p130 Rb, associated with cell-cycle arrest, indicating cell-cycle inhibition. Next, the effect of Acanthamoeba on retinoblastoma protein (pRb) phosphorylation was determined. pRb is a potent inhibitor of G 1 -to-S cell-cycle progression; however, its function is inhibited upon phosphorylation, allowing progression into S phase. Western blotting revealed that Acanthamoeba abolished pRb phosphorylation leading to cell-cycle arrest at the G 1 -to-S transition. Taken together, these studies demonstrated for the first time that Acanthamoeba inhibits the host cell cycle at the transcriptional level, as well as by modulating pRb phosphorylation using host cell-signalling mechanisms. A complete understanding of Acanthamoeba-host cell interactions may help in developing novel strategies to treat Acanthamoeba infections. INTRODUCTIONAcanthamoeba are the causative agents of life-threatening granulomatous amoebic encephalitis (GAE) in immunocompromised patients (Jones et al., 1975;Martinez, 1987;Di Gregorio et al., 1992;Friedland et al., 1992;Gonzalez et al., 1986;Gordon et al., 1992) and a common amoebic keratitis, a sight-threatening disease of the eye, which is mostly associated with contact-lens use (Chynn et al., 1995;Moore & McCulley, 1989;Wright et al., 1985; Mathers et al., 1996;Niederkorn et al., 1999;Marciano-Cabral & Cabral, 2003;Khan, 2003). In addition, Acanthamoeba have been associated with cutaneous lesions and sinusitis in immunocompromised patients (reviewed by Marciano-Cabral & Cabral, 2003). Over the past few decades, Acanthamoeba infections have remained significant and are on the rise. This is due to increasing populations of contact-lens wearers and immunocompromised patients, as well as improved awareness and detection of these emerging diseases. However, the precise mechanisms associated with the pathogenesis and pathophysiology of Acanthamoeba infection remain unclear. The clinical outcome of Acanthamoeba infection is dependent on the virulence of...

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Substance P-induced cadherin expression and its signal transduction in a cloned human corneal epithelial cell line

Cited by 70 publications

References 23 publications

Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

Cellular and nerve regeneration within a biosynthetic extracellular matrix for corneal transplantation

Acanthamoeba induces cell-cycle arrest in host cells

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