Maintenance of Vascular Endothelial Cell-Specific Properties after Immortalization with an Amphotrophic Replication-Deficient Retrovirus Containing Human Papilloma Virus 16 E6/E7 DNA

Fontijn, Ruud D.; Hop, C.; Brinkman, Herm Jan M.; Slater, Rosalyn; Westerveld, A.; Mourik, Jan A. van; Pannekoek, Hans

doi:10.1006/excr.1995.1025

Cited by 94 publications

(68 citation statements)

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“…At confluence cells were trypsinized and divided into two groups. One group was allowed to grow and was subsequently frozen while the other group was immortalized with replication-deficient retrovirus containing human papilloma virus 16 E6/E7 (Fontijn et al, 1995). All cell lines were stained with RPE65 a marker of RPE cells.…”

Section: Rpe Isolationmentioning

confidence: 99%

Subtype specific estrogen receptor action protects against changes in MMP-2 activation in mouse retinal pigmented epithelial cells

Elliot

Catanuto

Fernández

et al. 2008

Experimental Eye Research

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Eyes with age-related macular degeneration (AMD) demonstrate accumulation of specific deposits and extracellular matrix (ECM) molecules under the retinal pigment epithelium (RPE). AMD is about two times more prevalent in aging postmenopausal women. Therefore we studied whether 17β-estradiol (E 2 ) modulates the expression and activity of the trimolecular complex (MMP-2, TIMP-2 and MMP-14), molecules which are of major importance for ECM turnover in RPE. We used cell lines isolated from estrogen receptor knockout mice (ERKO) to determine which ER (estrogen receptor) subtype was important for ECM regulation in RPE cells.We found that mouse RPE sheets had higher baseline MMP-2 activity in the presence of ERβ. This correlated with higher MMP-2 activity in RPE cell lines isolated from ERKOα mice. Exposure to E 2 increased MMP-2 activity in mouse RPE cell lines. In addition E 2 increased transcriptional activation of the MMP-2 promoter through a functional Sp1 site which required the presence of ERβ, but not ERα. E 2 also maintained levels of pro MMP-2, and MMP-14 and TIMP-2 activity after oxidant injury. Since the direct effects of E 2 on MMP-2 transcriptional activation and the regulation of the trimolecular complex after oxidant induced injury requires ERβ, this receptor subtype may have a role as a potential therapeutic target to prevent changes in activation of MMP-2.

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Section: Rpe Isolationmentioning

confidence: 99%

Subtype specific estrogen receptor action protects against changes in MMP-2 activation in mouse retinal pigmented epithelial cells

Elliot

Catanuto

Fernández

et al. 2008

Experimental Eye Research

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“…Primary cells were split in a 1:3 ratio and used up to the sixth passage. An EC line, HM2, (R. Hofer-Warbinek, personal communication) was generated by infection with an amphotrophic helper-free recombinant retrovirus construct, pLXSN16 E6/E7, containing the E6/E7 open reading frame of human papilloma virus 16 (Fontijn et al, 1995) and cultured under selection of 100 µg/ml G418 (Sigma). Confluent monolayers were left for 48 hours and subsequent experiments were done without changing the cell culture medium.…”

Section: Cell Culturementioning

confidence: 99%

HERC5, a HECT E3 ubiquitin ligase tightly regulated in LPS activated endothelial cells

Kroismayr

Baranyi

Stehlik

et al. 2004

Journal of Cell Science

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By differential screening we isolated genes upregulated in inflammatory cytokine-stimulated human skin microvascular endothelial cells. One of these cDNAs encoded RCC1 (regulator of chromosome condensation 1)-like repeats and a HECT (homologous to E6-AP C-terminus) domain, representing a member of the HERC (HECT and RCC1 domain protein) family of ubiquitin ligases. The mRNA level of this member, HERC5, is specifically upregulated in endothelial cells by the pro-inflammatory cytokines tumor necrosis factor α and interleukin 1β, and by lipopolysaccharide (LPS), but is hardly expressed in other cells of the vascular wall such as primary smooth muscle cells and fibroblasts. Regulation of HERC5 gene expression suggests a critical role for the transcription factor NF-κB. In contrast to mRNA expression HERC5 protein is subject of enhanced degradation upon LPS stimulation of endothelial cells. The time course of LPS-induced changes in HERC5 protein and mRNA levels suggests that the initial drop in HERC5 protein is balanced by increased protein synthesis due to upregulation of HERC5 mRNA. This leads to recovery of HERC5 protein levels within 12 hours of LPS stimulation and points at a tight control of HERC5 protein. To analyze functional activity of this putative member of the ubiquitin-conjugating pathway we performed in vitro assays with different ubiquitin-conjugating enzymes. We found that HERC5 possesses ubiquitin ligase activity and requires the presence of the ubiquitin-conjugating enzyme UbcH5a for its activity. These data show for the first time that a functionally active HECT ubiquitin ligase exhibits a tightly controlled cytosolic level under inflammatory conditions in endothelial cells.

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“…TNF Bioassay-10 4 TNF-responsive endothelial cells (ECRF-24 cells) (16) were seeded in a 96-well plate in IMDM containing 5% FCS (Bodinco), 100 units/ml penicillin, 100 g/ml streptomycin, 50 M 2-mercaptoethanol, 20 g/ml human apo-transferrin (Sigma). After 24 h of incubation, the samples containing TNF, adalimumab, and monoclonal antibody were added.…”

Section: Generation Of F(abј) 2 and Fab Fragments-f(abј)mentioning

confidence: 99%

Functional Analysis of the Anti-adalimumab Response Using Patient-derived Monoclonal Antibodies

Schouwenburg

Kruithof

Votsmeier

et al. 2014

Journal of Biological Chemistry

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Background: Therapeutic antibodies such as adalimumab can elicit anti-drug antibodies. Results: Monoclonal patient-derived antibodies were generated and found to compete for binding to adalimumab and are neutralizing, but they show markedly different fine-specificity. Conclusion: Anti-adalimumab antibodies bind to overlapping but distinct epitopes on adalimumab. Significance: Even for a fully human therapeutic antibody, there may be multiple determinants that contribute to immunogenicity.

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Maintenance of Vascular Endothelial Cell-Specific Properties after Immortalization with an Amphotrophic Replication-Deficient Retrovirus Containing Human Papilloma Virus 16 E6/E7 DNA

Cited by 94 publications

References 0 publications

Subtype specific estrogen receptor action protects against changes in MMP-2 activation in mouse retinal pigmented epithelial cells

Subtype specific estrogen receptor action protects against changes in MMP-2 activation in mouse retinal pigmented epithelial cells

HERC5, a HECT E3 ubiquitin ligase tightly regulated in LPS activated endothelial cells

Functional Analysis of the Anti-adalimumab Response Using Patient-derived Monoclonal Antibodies

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