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Perforin competent CD8 T cells are sufficient to cause immune-mediated blood-brain barrier disruption.

Johnson HL, Willenbring RC, Jin F, Manhart WA, LaFrance SJ, Pirko I, Johnson AJ - PLoS ONE (2014)

Bottom Line: However, the underlying contribution of inflammatory-derived factors leading to pathology associated with blood-brain barrier (BBB) disruption remains poorly understood.To investigate this, we isolated the CD8 T cell as the sole perforin-expressing cell type in the PIFS model through adoptive transfer techniques.Therefore, CD8 T cells are sufficient as a sole perforin-expressing cell type to cause BBB disruption in the PIFS model.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Mayo Clinic, Rochester, Minnesota, United States of America; Department of Immunology, Mayo Clinic, Rochester, Minnesota, United States of America; Neurobiology of Disease Graduate Program, Mayo Graduate School, Rochester, Minnesota, United States of America.

ABSTRACT
Numerous neurological disorders are characterized by central nervous system (CNS) vascular permeability. However, the underlying contribution of inflammatory-derived factors leading to pathology associated with blood-brain barrier (BBB) disruption remains poorly understood. In order to address this, we developed an inducible model of BBB disruption using a variation of the Theiler's murine encephalomyelitis virus (TMEV) model of multiple sclerosis. This peptide induced fatal syndrome (PIFS) model is initiated by virus-specific CD8 T cells and results in severe CNS vascular permeability and death in the C57BL/6 mouse strain. While perforin is required for BBB disruption, the cellular source of perforin has remained unidentified. In addition to CD8 T cells, various innate immune cells also express perforin and therefore could also contribute to BBB disruption. To investigate this, we isolated the CD8 T cell as the sole perforin-expressing cell type in the PIFS model through adoptive transfer techniques. We determined that C57BL/6 perforin-/- mice reconstituted with perforin competent CD8 T cells and induced to undergo PIFS exhibited: 1) heightened CNS vascular permeability, 2) increased astrocyte activation as measured by GFAP expression, and 3) loss of linear organization of BBB tight junction proteins claudin-5 and occludin in areas of CNS vascular permeability when compared to mock-treated controls. These results are consistent with the characteristics associated with PIFS in perforin competent mice. Therefore, CD8 T cells are sufficient as a sole perforin-expressing cell type to cause BBB disruption in the PIFS model.

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Increased astrocyte activation colocalizes with CNS vascular permeability post-induction of PIFS in perforin−/− mice reconstituted with perforin competent CD8 T cells.Representative confocal microscopic images illustrating astrocyte expression of GFAP in (A) E7 peptide-treated (n = 4) and (B) VP2121–130 peptide-treated (n = 6) perforin−/− mice reconstituted with perforin competent CD8 T cells. Mice treated with (B) VP2121–130 peptide displayed increased astrocyte activation, as measured by heightened GFAP expression, in areas of CNS vascular permeability when compared to (A) mock E7 peptide-treated negative control mice. CNS permeability is determined by leakage of intravenously administered FITC-albumin.
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pone-0111401-g003: Increased astrocyte activation colocalizes with CNS vascular permeability post-induction of PIFS in perforin−/− mice reconstituted with perforin competent CD8 T cells.Representative confocal microscopic images illustrating astrocyte expression of GFAP in (A) E7 peptide-treated (n = 4) and (B) VP2121–130 peptide-treated (n = 6) perforin−/− mice reconstituted with perforin competent CD8 T cells. Mice treated with (B) VP2121–130 peptide displayed increased astrocyte activation, as measured by heightened GFAP expression, in areas of CNS vascular permeability when compared to (A) mock E7 peptide-treated negative control mice. CNS permeability is determined by leakage of intravenously administered FITC-albumin.

Mentions: Activation of astrocytes, as measured by GFAP expression, co-localizes with CNS vascular permeability in C57BL/6 mice induced to undergo PIFS [18]. To determine the extent astrocytes were activated in perforin−/− mice reconstituted with perforin competent CD8 T cells, we stained for astrocyte expression of GFAP and performed confocal microscopy. We found that mice treated with VP2121–130 peptide displayed an increase in astrocyte size consistent with astrogliosis (Figure 3B), when compared to E7 peptide-treated mice (Figure 3A). We also observed colocalization of increased astrocyte activation with CNS vascular permeability. This is consistent with previous studies in which PIFS was evaluated in perforin competent wild-type C57BL/6 mice [18]. In this study, in accordance with previous studies, there is a clear increase in astrocyte activation in areas with vascular leak as compared to unaffected tissue (Figure 3B).


Perforin competent CD8 T cells are sufficient to cause immune-mediated blood-brain barrier disruption.

Johnson HL, Willenbring RC, Jin F, Manhart WA, LaFrance SJ, Pirko I, Johnson AJ - PLoS ONE (2014)

Increased astrocyte activation colocalizes with CNS vascular permeability post-induction of PIFS in perforin−/− mice reconstituted with perforin competent CD8 T cells.Representative confocal microscopic images illustrating astrocyte expression of GFAP in (A) E7 peptide-treated (n = 4) and (B) VP2121–130 peptide-treated (n = 6) perforin−/− mice reconstituted with perforin competent CD8 T cells. Mice treated with (B) VP2121–130 peptide displayed increased astrocyte activation, as measured by heightened GFAP expression, in areas of CNS vascular permeability when compared to (A) mock E7 peptide-treated negative control mice. CNS permeability is determined by leakage of intravenously administered FITC-albumin.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206459&req=5

pone-0111401-g003: Increased astrocyte activation colocalizes with CNS vascular permeability post-induction of PIFS in perforin−/− mice reconstituted with perforin competent CD8 T cells.Representative confocal microscopic images illustrating astrocyte expression of GFAP in (A) E7 peptide-treated (n = 4) and (B) VP2121–130 peptide-treated (n = 6) perforin−/− mice reconstituted with perforin competent CD8 T cells. Mice treated with (B) VP2121–130 peptide displayed increased astrocyte activation, as measured by heightened GFAP expression, in areas of CNS vascular permeability when compared to (A) mock E7 peptide-treated negative control mice. CNS permeability is determined by leakage of intravenously administered FITC-albumin.
Mentions: Activation of astrocytes, as measured by GFAP expression, co-localizes with CNS vascular permeability in C57BL/6 mice induced to undergo PIFS [18]. To determine the extent astrocytes were activated in perforin−/− mice reconstituted with perforin competent CD8 T cells, we stained for astrocyte expression of GFAP and performed confocal microscopy. We found that mice treated with VP2121–130 peptide displayed an increase in astrocyte size consistent with astrogliosis (Figure 3B), when compared to E7 peptide-treated mice (Figure 3A). We also observed colocalization of increased astrocyte activation with CNS vascular permeability. This is consistent with previous studies in which PIFS was evaluated in perforin competent wild-type C57BL/6 mice [18]. In this study, in accordance with previous studies, there is a clear increase in astrocyte activation in areas with vascular leak as compared to unaffected tissue (Figure 3B).

Bottom Line: However, the underlying contribution of inflammatory-derived factors leading to pathology associated with blood-brain barrier (BBB) disruption remains poorly understood.To investigate this, we isolated the CD8 T cell as the sole perforin-expressing cell type in the PIFS model through adoptive transfer techniques.Therefore, CD8 T cells are sufficient as a sole perforin-expressing cell type to cause BBB disruption in the PIFS model.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Mayo Clinic, Rochester, Minnesota, United States of America; Department of Immunology, Mayo Clinic, Rochester, Minnesota, United States of America; Neurobiology of Disease Graduate Program, Mayo Graduate School, Rochester, Minnesota, United States of America.

ABSTRACT
Numerous neurological disorders are characterized by central nervous system (CNS) vascular permeability. However, the underlying contribution of inflammatory-derived factors leading to pathology associated with blood-brain barrier (BBB) disruption remains poorly understood. In order to address this, we developed an inducible model of BBB disruption using a variation of the Theiler's murine encephalomyelitis virus (TMEV) model of multiple sclerosis. This peptide induced fatal syndrome (PIFS) model is initiated by virus-specific CD8 T cells and results in severe CNS vascular permeability and death in the C57BL/6 mouse strain. While perforin is required for BBB disruption, the cellular source of perforin has remained unidentified. In addition to CD8 T cells, various innate immune cells also express perforin and therefore could also contribute to BBB disruption. To investigate this, we isolated the CD8 T cell as the sole perforin-expressing cell type in the PIFS model through adoptive transfer techniques. We determined that C57BL/6 perforin-/- mice reconstituted with perforin competent CD8 T cells and induced to undergo PIFS exhibited: 1) heightened CNS vascular permeability, 2) increased astrocyte activation as measured by GFAP expression, and 3) loss of linear organization of BBB tight junction proteins claudin-5 and occludin in areas of CNS vascular permeability when compared to mock-treated controls. These results are consistent with the characteristics associated with PIFS in perforin competent mice. Therefore, CD8 T cells are sufficient as a sole perforin-expressing cell type to cause BBB disruption in the PIFS model.

Show MeSH
Related in: MedlinePlus