Es are in constant physical contact with the EC surface. Additionally, in the brains of both mice and human with CM, leukocytes (monocytes and 
T cells) become arrested in brain microvessels [2] providing further means for intimate EC/T cell interactions. It has long been established that CM is a T cell-dependent disease [41,42], with both CD4+ and CD8+ T cells playing key roles in CM pathogenesis [43,44]. Moreover, this cell-cell contact plays an important role in brain endothelial activation [45], as assessed notably by a dramatic increase in plasma levels endothelial microparticles at the time ofCM [46]. The data presented here, in combination with our recent demonstration that HBEC can transfer antigens from malarial-infected red blood cells onto their surface, buy Fruquintinib thereby becoming a target for the immune response, provide key evidence for HBEC to act as antigen presenting cells with the presentation of malaria antigens by brain EC to T cells and the potential activation of cytotoxic mechanisms providing a new explanation for CM pathogenesis.Supporting Informationreduction in both CD4+ and CD8+ T cell proliferation. Graphical representation of fold increase in proliferation of aCD3/CD28 stimulated CD4+ and CD8+ T cells co-cultured with TNF/IFNc stimulated HBEC over unstimulated (control) CD4+ and CD8+ T cell proliferation. Proliferation assessed by CFSE following 6 days of co-culture either in 24 well plates (black bars) or in 0.4 mm transwells (white bars). (TIF)Figure S1 Separation of HBEC and PBMC results in aAcknowledgmentsWe thank Gerard Chan for his technical assistance.Author ContributionsConceived and designed the GNF-7 web experiments: JW VC GG. Performed the experiments: JW 
SO. Analyzed the data: JW SO. Contributed reagents/ materials/analysis tools: PC. Wrote the paper: JW VC GG.
Nucleic acids are 23977191 highly polymorphic: depending on the sequences and environmental conditions they may exist in a variety of secondary structures such as duplexes, triplexes, tetraplexes, bulges, hairpins, loops [1,2]. Such non-canonical structures in nucleic acids are of general biological significance: they have been postulated to mediate protein-nucleic acid interactions, either by contacting protein residues directly or by producing a distinct tertiary structure to which the protein binds [3], and to function as intermediates in the generation of frameshift mutations when errors in DNA replication occur [4,5]. In particular, extra-helical bases are thought to be implicated in nucleic acid non-canonical functions [6]. An essentially unlimited combination of secondary structural elements has been extensively described in RNA, where the single-stranded (ss) nucleic acid folds back on itself; however, DNA can also produce complex secondary structures during replication and recombination [7]. A shift of the reading frame during template-dependent DNA synthesis can lead to the addition or deletion of one or more nucleotide residues (nts) in the newly synthesized DNA, ensuing in bulged or mismatched structures. Bulged bases derived from replicative errors are considered the first step of frame-shift mutagenesis [6], resultingin a variety of diseases and cancers (e.g., myotonic dystrophy, Huntington’s disease, Friederich’s ataxia, and fragile X syndrome). In general, compounds capable of binding to non-canonical conformations of the DNA could have significant therapeutic potential. Several derivatives with unrelated structures have been reported to individually targ.Es are in constant physical contact with the EC surface. Additionally, in the brains of both mice and human with CM, leukocytes (monocytes and T cells) become arrested in brain microvessels [2] providing further means for intimate EC/T cell interactions. It has long been established that CM is a T cell-dependent disease [41,42], with both CD4+ and CD8+ T cells playing key roles in CM pathogenesis [43,44]. Moreover, this cell-cell contact plays an important role in brain endothelial activation [45], as assessed notably by a dramatic increase in plasma levels endothelial microparticles at the time ofCM [46]. The data presented here, in combination with our recent demonstration that HBEC can transfer antigens from malarial-infected red blood cells onto their surface, thereby becoming a target for the immune response, provide key evidence for HBEC to act as antigen presenting cells with the presentation of malaria antigens by brain EC to T cells and the potential activation of cytotoxic mechanisms providing a new explanation for CM pathogenesis.Supporting Informationreduction in both CD4+ and CD8+ T cell proliferation. Graphical representation of fold increase in proliferation of aCD3/CD28 stimulated CD4+ and CD8+ T cells co-cultured with TNF/IFNc stimulated HBEC over unstimulated (control) CD4+ and CD8+ T cell proliferation. Proliferation assessed by CFSE following 6 days of co-culture either in 24 well plates (black bars) or in 0.4 mm transwells (white bars). (TIF)Figure S1 Separation of HBEC and PBMC results in aAcknowledgmentsWe thank Gerard Chan for his technical assistance.Author ContributionsConceived and designed the experiments: JW VC GG. Performed the experiments: JW SO. Analyzed the data: JW SO. Contributed reagents/ materials/analysis tools: PC. Wrote the paper: JW VC GG.
Nucleic acids are 23977191 highly polymorphic: depending on the sequences and environmental conditions they may exist in a variety of secondary structures such as duplexes, triplexes, tetraplexes, bulges, hairpins, loops [1,2]. Such non-canonical structures in nucleic acids are of general biological significance: they have been postulated to mediate protein-nucleic acid interactions, either by contacting protein residues directly or by producing a distinct tertiary structure to which the protein binds [3], and to function as intermediates in the generation of frameshift mutations when errors in DNA replication occur [4,5]. In particular, extra-helical bases are thought to be implicated in nucleic acid non-canonical functions [6]. An essentially unlimited combination of secondary structural elements has been extensively described in RNA, where the single-stranded (ss) nucleic acid folds back on itself; however, DNA can also produce complex secondary structures during replication and recombination [7]. A shift of the reading frame during template-dependent DNA synthesis can lead to the addition or deletion of one or more nucleotide residues (nts) in the newly synthesized DNA, ensuing in bulged or mismatched structures. Bulged bases derived from replicative errors are considered the first step of frame-shift mutagenesis [6], resultingin a variety of diseases and cancers (e.g., myotonic dystrophy, Huntington’s disease, Friederich’s ataxia, and fragile X syndrome). In general, compounds capable of binding to non-canonical conformations of the DNA could have significant therapeutic potential. Several derivatives with unrelated structures have been reported to individually targ.