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Pro-inflammatory activation changes intracellular transport of bevacizumab in the retinal pigment epithelium in vitro.
Hildebrandt, Julia; Käckenmeister, Tom; Winkelmann, Katrin; Dörschmann, Philipp; Roider, Johann; Klettner, Alexa.
Afiliação
  • Hildebrandt J; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany.
  • Käckenmeister T; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany.
  • Winkelmann K; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany.
  • Dörschmann P; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany.
  • Roider J; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany.
  • Klettner A; Department of Ophthalmology, University Medical Center, University of Kiel, Quincke Research Center, Rosalind-Franklin-Str. 9, 24105, Kiel, Germany. AlexaKarina.Klettner@uksh.de.
Graefes Arch Clin Exp Ophthalmol ; 260(3): 857-872, 2022 Mar.
Article em En | MEDLINE | ID: mdl-34643794
ABSTRACT

PURPOSE:

Bevacizumab is taken up and transported through the retinal pigment epithelium. Inflammatory signaling may influence this interaction. In the present study, we have investigated the effect of pro-inflammatory stimuli on the uptake, intracellular localization, and transepithelial transport of bevacizumab.

METHODS:

ARPE-19 cell line or primary porcine RPE cells were treated with clinical relevant concentrations of bevacizumab (250 µg/ml). Pro-inflammatory signaling was induced by TLR-3 agonist polyinosinicpolycytidylic acid (Poly IC). Viability was investigated with MTT and trypan-blue exclusion assay, and cell number, uptake, and intracellular localization were investigated with immunofluorescence, investigating also actin filaments, the motor protein myosin 7a and lysosomes. Immunofluorescence signals were quantified. Intracellular bevacizumab was additionally detected in Western blot. Barrier function was investigated with transepithelial resistant measurements (TER). The transepithelial transport of bevacizumab and its influence on cytokine (IL-6, IL-8, IL-1ß, TNFα) secretion was investigated with ELISA.

RESULTS:

Poly IC in combination with bevacizumab reduced the viability of the cells. Treatment with Poly IC reduced the uptake of bevacizumab, changed the intensity of the actin filaments, and reduced the colocalization with myosin 7a. In addition, Poly IC reduced the capacity of RPE cells to transport bevacizumab over the barrier. In addition, bevacizumab reduced the secretion of IL-8 and TNFα after Poly IC stimulation at selected time points.

CONCLUSIONS:

Pro-inflammatory activation of RPE cells with TLR-3 agonist Poly IC changes the interaction of RPE cells with the anti-VEGF compound bevacizumab, reducing its uptake and transport. On the other hand, bevacizumab might influence pro-inflammatory cytokine release. Our data indicate that inflammation may influence the pharmacokinetic of bevacizumab in the retina.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Inibidores da Angiogênese / Epitélio Pigmentado da Retina Limite: Animals Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Inibidores da Angiogênese / Epitélio Pigmentado da Retina Limite: Animals Idioma: En Ano de publicação: 2022 Tipo de documento: Article