Abstract
While there are murine and rat choroid plexus epithelial cell cultures, a translationally relevant model for choroid plexus activation and function is still lacking. The rhesus macaque is the gold standard for modeling viral infection and activation of CNS, including HIV-associated neurocognitive disorders. We have developed a rhesus macaque choroid plexus epithelial cell culture model which we believe to be suitable for studies of inflammation associated with viral infection of the CNS. Epithelial morphology and function were assessed using vimentin, phalloidin, the tight junction protein zonula-occludens-1 (ZO-1), and focal adhesion kinase (FAK). Choroid plexus epithelial cell type was confirmed using immunofluorescence with two proteins highly expressed in the choroid plexus: transthyretin and α-klotho. Finally, barrier properties of the model were monitored using pro- and anti-inflammatory mediators (TNF-α, the TLR2 agonist PamCys3K, and dexamethasone). When pro-inflammatory TNF-α was added to the xCelligence wells, there was a decrease in barrier function, which decreased in a step-wise fashion with each additional administration. This barrier function was repaired upon addition of the steroid dexamethasone. The TLR2 agonist PAM3CysK increased barrier functions in TNF-α treated wells. We have presented a model of the blood-CSF barrier that will allow study into pro- and anti-inflammatory conditions in the brain, while simultaneously measuring real time changes to epithelial cells.
Highlights
The choroid plexus, comprising the blood-cerebrospinal fluid (CSF) barrier, lines the lateral, 3rd and 4th ventricles of the brain (Zheng and Zhao, 2002; Monnot and Zheng, 2013)
Collagen appeared to improve the initial plating of primary choroid plexus cultures, choroid plexus epithelial cells grew as monolayers on either gelatin or collagen coated plates
As HIV does not have a translationally relevant small animal model, we believed it was critical to develop a choroid plexus cell culture model in a species that does allow studies of HIV neuropathology: the rhesus macaque (Ivey et al, 2009a). We believe this model will be useful for the study of accelerated aging or “inflammaging,” as the choroid plexus is the main source of α-klotho, the “anti-aging hormone,” in the brain (Kuro-o et al, 1997; Takahashi et al, 2000; German et al, 2012)
Summary
The choroid plexus, comprising the blood-cerebrospinal fluid (CSF) barrier, lines the lateral, 3rd and 4th ventricles of the brain (Zheng and Zhao, 2002; Monnot and Zheng, 2013). It is responsible for secreting CSF, allowing the passive diffusion of water and oxygen into the brain, and facilitating the active transport of glucose and larger particles into the brain (Zheng and Zhao, 2002; Monnot and Zheng, 2013). There are unique populations of macrophages that line the endothelial blood vessels and choroid plexus resident macrophages (Bragg et al, 2002; Kim et al, 2006; Delery and Maclean, 2018).
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