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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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CD8 T cells as sole perforin wielding source are sufficient to induce CNS vascular permeability.The extent of CNS vascular permeability is illustrated by raw image and 3D transparency rendering of gadolinium-enhancing areas from T1-weighted MRI scans in mice administered (A,C) mock E7 control peptide or (B,D) VP2121–130 peptide. MRI images are separated to show both methods of CD8+ T cell selection (A,B) negative sort or (C,D) positive sort to isolate and transfer CD8+ cells. (E) Quantification of the 3D volume of vascular leakage from all mice (both negative and positive sort) revealed that VP2121–130 peptide-treated mice having CD8 T cells as the sole perforin-expressing cell type have a significantly more amount of gadolinium leakage (p<0.05) (n = 5,9 per group). (F) A significant increase in CNS vascular permeability detectable by intravenously injected rhodamine dextran accumulation in brain homogenates was observed in VP2121–130 peptide-treated animals (n = 6) compared to mock E7 peptide-treated animals (n = 4) (p<0.05). Results are depicted as mean ± SEM.
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pone-0111401-g002: CD8 T cells as sole perforin wielding source are sufficient to induce CNS vascular permeability.The extent of CNS vascular permeability is illustrated by raw image and 3D transparency rendering of gadolinium-enhancing areas from T1-weighted MRI scans in mice administered (A,C) mock E7 control peptide or (B,D) VP2121–130 peptide. MRI images are separated to show both methods of CD8+ T cell selection (A,B) negative sort or (C,D) positive sort to isolate and transfer CD8+ cells. (E) Quantification of the 3D volume of vascular leakage from all mice (both negative and positive sort) revealed that VP2121–130 peptide-treated mice having CD8 T cells as the sole perforin-expressing cell type have a significantly more amount of gadolinium leakage (p<0.05) (n = 5,9 per group). (F) A significant increase in CNS vascular permeability detectable by intravenously injected rhodamine dextran accumulation in brain homogenates was observed in VP2121–130 peptide-treated animals (n = 6) compared to mock E7 peptide-treated animals (n = 4) (p<0.05). Results are depicted as mean ± SEM.

Mentions: In order to quantitatively evaluate the full volume of CNS vascular leak in animals undergoing PIFS, where CD8 T cells were the sole perforin-expressing cell type, we performed volumetric analysis of 3D gadolinium-enhanced T1-weighted MRI scans using Analyze 11.0 software developed by Mayo Clinic's Biomedical Imaging Resource [30], [31]. Quantification of the 3D volume of gadolinium leakage from vasculature revealed that VP2121–130 peptide-treated mice reconstituted with perforin competent CD8 T cells had increased CNS vascular permeability (Figure 2 B,D) when compared to mock E7 peptide-treated controls (Figure 2 A,C) (p<0.05, Figure 2E). CD8+ cells were selected using a negative (Figure 2 A,B) or positive (Figure 2 C,D) sort method. Importantly, an additional VP2121–130 peptide-treated mouse, not included in this data set, was moribund and unable to be scanned. Post-mortem analysis revealed that this mouse had the highest transfer of CD8 T cells (data not shown).


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)

CD8 T cells as sole perforin wielding source are sufficient to induce CNS vascular permeability.The extent of CNS vascular permeability is illustrated by raw image and 3D transparency rendering of gadolinium-enhancing areas from T1-weighted MRI scans in mice administered (A,C) mock E7 control peptide or (B,D) VP2121–130 peptide. MRI images are separated to show both methods of CD8+ T cell selection (A,B) negative sort or (C,D) positive sort to isolate and transfer CD8+ cells. (E) Quantification of the 3D volume of vascular leakage from all mice (both negative and positive sort) revealed that VP2121–130 peptide-treated mice having CD8 T cells as the sole perforin-expressing cell type have a significantly more amount of gadolinium leakage (p<0.05) (n = 5,9 per group). (F) A significant increase in CNS vascular permeability detectable by intravenously injected rhodamine dextran accumulation in brain homogenates was observed in VP2121–130 peptide-treated animals (n = 6) compared to mock E7 peptide-treated animals (n = 4) (p<0.05). Results are depicted as mean ± SEM.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0111401-g002: CD8 T cells as sole perforin wielding source are sufficient to induce CNS vascular permeability.The extent of CNS vascular permeability is illustrated by raw image and 3D transparency rendering of gadolinium-enhancing areas from T1-weighted MRI scans in mice administered (A,C) mock E7 control peptide or (B,D) VP2121–130 peptide. MRI images are separated to show both methods of CD8+ T cell selection (A,B) negative sort or (C,D) positive sort to isolate and transfer CD8+ cells. (E) Quantification of the 3D volume of vascular leakage from all mice (both negative and positive sort) revealed that VP2121–130 peptide-treated mice having CD8 T cells as the sole perforin-expressing cell type have a significantly more amount of gadolinium leakage (p<0.05) (n = 5,9 per group). (F) A significant increase in CNS vascular permeability detectable by intravenously injected rhodamine dextran accumulation in brain homogenates was observed in VP2121–130 peptide-treated animals (n = 6) compared to mock E7 peptide-treated animals (n = 4) (p<0.05). Results are depicted as mean ± SEM.
Mentions: In order to quantitatively evaluate the full volume of CNS vascular leak in animals undergoing PIFS, where CD8 T cells were the sole perforin-expressing cell type, we performed volumetric analysis of 3D gadolinium-enhanced T1-weighted MRI scans using Analyze 11.0 software developed by Mayo Clinic's Biomedical Imaging Resource [30], [31]. Quantification of the 3D volume of gadolinium leakage from vasculature revealed that VP2121–130 peptide-treated mice reconstituted with perforin competent CD8 T cells had increased CNS vascular permeability (Figure 2 B,D) when compared to mock E7 peptide-treated controls (Figure 2 A,C) (p<0.05, Figure 2E). CD8+ cells were selected using a negative (Figure 2 A,B) or positive (Figure 2 C,D) sort method. Importantly, an additional VP2121–130 peptide-treated mouse, not included in this data set, was moribund and unable to be scanned. Post-mortem analysis revealed that this mouse had the highest transfer of CD8 T cells (data not shown).

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