Calnexin is necessary for T cell transmigration into the central nervous system

Jung, Joanna; Eggleton, Paul; Robinson, Alison; Wang, Jessica; Gutowski, N. J.; Holley, Janet E.; Newcombe, Jia; Dudek, Elzbieta; Paul, Amber M.; Zochodne, Douglas W.; Kraus, Allison; Power, Christopher; Agellon, Luis B.; Michalak, Marek

doi:10.1172/jci.insight.98410

Cited by 15 publications

(39 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Male and female Canx-deficient mice (Canx −/− ) and Canx −/− transgenic mice reconstituted with full length recombinant Canx by transgene expression (designated Tg-CanxFL) were generated as previously described. 15,16 An expression vector designed for ubiquitous expression under the ROSA26 promoter was used to generate Canx −/− transgenic mice expressing cytoplasmic C-tail truncated form of Canx protein (designated Tg-CanxΔC). The vector encoded amino acids 1-505 of the fulllength Canx corresponding to ER lumenal and transmembrane domains of the protein followed by 10 amino acids C-terminal extension containing the ER retention signal (Figure 2A).…”

Section: Mouse Modelsmentioning

confidence: 99%

“…The bEND.3 cell line (male cells) was purchased from ATCC (ATCC CRL-2299) and cultured under recommended conditions as previously described. 16 Disruption of the Fabp5 gene in the bEND.3 cell line was achieved with two paired sgRNAs designed to excise a large part of the gene. sgRNAs flanking Exon 1 in Fabp5 gene ( Figure S1) were identified using the web-based tool: https://crispr.mit.edu/ (retrieved May 2020): sgRNA1 5′-CACAACTCCGGGCGTGGCGA-3′; sgRNA2 5′-TTTATCGCCGAACCCCTCCC-3′.…”

Section: Fabp5 −/− Bend3 Cell Linementioning

confidence: 99%

“…The sgRNA coding vector was created by ligating the sgRNA sequences into the SpCas9(BB)-2A-Puro (Px459) V2.0 plasmid [a gift from Feng Zhang (Addgene Plasmid #62988)] as previously described. 16 The assembled plasmid was transfected into wildtype bEND.3 cells using TurboFect Transfection reagent (Thermo Fisher Scientific) following the manufacturer's recommended protocol designed for adherent cells. Individual clones were picked and assayed by both PCR and immunoblot analysis, to confirm successful inactivation of the Fabp5 gene.…”

Section: Fabp5 −/− Bend3 Cell Linementioning

confidence: 99%

“…T-cell migration across endothelial bEND.3 monolayers was carried out as described previously. 16 Briefly, wild-type or genetically modified bEND.3 cells were plated onto 12-well plates to form an intact monolayer. The endothelial cell monolayer was treated for 4 hours with IL1ß (10 ng/mL, eBioscience) or 16 hours with TNFα (3 ng/mL) (Invitrogen) and IFNɣ (60 ng/ mL, eBioscience).…”

Section: Trans-endothelial Migration Analysismentioning

confidence: 99%

“…15 Surprisingly, these mice also show resistance to EAE induction which is due to the inability of activated T-cells to cross Canx-deficient blood-brain barrier endothelial cells. 16 During our search for proteins that interact with the cytoplasmic C-tail domain of Canx, we identified Fabp5 as a brain-expressed protein showing high-affinity and stable binding. 17 Because of the interaction of these two proteins and considering the noteworthy phenotypic similarity of mice lacking either Fabp5 or Canx in showing resistance to EAE, we tested the hypothesis that interaction between these two proteins is central to EAE induction in mice.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

The Fabp5/calnexin complex is a prerequisite for sensitization of mice to experimental autoimmune encephalomyelitis

et al. 2020

Self Cite

View full text Add to dashboard Cite

We previously showed that calnexin (Canx)-deficient mice are desensitized to experimental autoimmune encephalomyelitis (EAE) induction, a model that is frequently used to study inflammatory demyelinating diseases, due to increased resistance of the blood-brain barrier to immune cell transmigration. We also discovered that Fabp5, an abundant cytoplasmic lipid-binding protein found in brain endothelial cells, makes protein-protein contact with the cytoplasmic C-tail domain of Canx. Remarkably, both Canx-deficient and Fabp5-deficient mice commonly manifest resistance to EAE induction. Here, we evaluated the importance of Fabp5/Canx interactions on EAE pathogenesis and on the patency of a model blood-brain barrier to T-cell transcellular migration. The results demonstrate that formation of a complex comprised of Fabp5 and the C-tail domain of Canx dictates the permeability of the model blood-brain barrier to immune cells and is also a prerequisite for EAE pathogenesis.

show abstract

Section: Mouse Modelsmentioning

confidence: 99%

Section: Fabp5 −/− Bend3 Cell Linementioning

confidence: 99%

Section: Fabp5 −/− Bend3 Cell Linementioning

confidence: 99%

Section: Trans-endothelial Migration Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The Fabp5/calnexin complex is a prerequisite for sensitization of mice to experimental autoimmune encephalomyelitis

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

A View of the Endoplasmic Reticulum Through the Calreticulin Lens

Agellon

Michalak

2021

Cellular Biology of the Endoplasmic Reticulum

View full text Add to dashboard Cite

Engagement of people with multiple sclerosis to enhance research into the physiological effect of hyperbaric oxygen therapy

Moore

Eggleton

Smerdon

et al. 2020

Multiple Sclerosis and Related Disorders

Self Cite

View full text Add to dashboard Cite

Background: Thousands of people with multiple sclerosis (MS) have used selfadministered oxygen therapy in the UK. Clinical trials have been performed, with scant evidence that people with MS have been consulted to explore how they benefit from or how to optimise this treatment. The conventional MS disease disability scores used in trials seldom reflect the effects individuals report when using oxygen therapy to treat their symptoms.Methods: Three people with MS and the manager of an MS Centre formed a public involvement group and collaborated with clinicians and scientists to inform a lab-based study to investigate the physiological effects of oxygen therapy on microvascular brain endothelial cells.Results: People with MS often use oxygen therapy at a later stage when their symptoms worsen and only after using other treatments. The frequency of oxygen therapy sessions and hyperbaric pressure is individualised and varies for people with MS. Despite direct comparisons of efficacy proving difficult, most individuals are exposed to 100% O2 at 1.5 atmosphere absolute (ATA; 1140 mmHg absolute) for 60 min. In a laboratory-based study human brain endothelial cells were exposed in vitro to 152 mmHg O2 for 60 min with and without pressure, as this equates to 20% O2 achievable via hyperbarics, which was then replicated at atmospheric pressure. A significant reduction in endothelial cells ICAM-1 (CD54) implicated in inflammatory cell margination across the blood brain barrier was observed under oxygen treatment. Conclusions:By collaborating with people living with MS, we were able to design laboratory-based experimental protocols that replicate their treatment regimens to advance our understanding of the physiological effects of hyperbaric oxygen treatment on brain cells and their role in neuroinflammation.

show abstract

Calnexin is necessary for T cell transmigration into the central nervous system

Cited by 15 publications

References 43 publications

The Fabp5/calnexin complex is a prerequisite for sensitization of mice to experimental autoimmune encephalomyelitis

The Fabp5/calnexin complex is a prerequisite for sensitization of mice to experimental autoimmune encephalomyelitis

A View of the Endoplasmic Reticulum Through the Calreticulin Lens

Engagement of people with multiple sclerosis to enhance research into the physiological effect of hyperbaric oxygen therapy

Contact Info

Product

Resources

About