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Flow cytometric characterization of brain dendritic cell subsets after murine stroke.

Pösel C, Uri A, Schulz I, Boltze J, Weise G, Wagner DC - Exp Transl Stroke Med (2014)

Bottom Line: Sterile inflammation is a substantial element of post-stroke pathophysiology with the determination of autoimmunity versus tolerance being one of its most important aspects.The latter could be differentiated into the classical migratory DC subpopulations (CD11b+ and CD103+), but no CD4 or CD8+ DC were found.Finally, stroke caused a significant increase of CD11b/CD103 double-positive DC in the affected brain hemisphere.

View Article: PubMed Central - PubMed

Affiliation: Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.

ABSTRACT

Background: Sterile inflammation is a substantial element of post-stroke pathophysiology with the determination of autoimmunity versus tolerance being one of its most important aspects. It is believed that this determination is initiated relatively early after stroke onset by clearing macrophages and migratory dendritic cells (DC). However, the phenotypic differentiation of macrophages and DC is intricate particularly in the disease context. Here, we utilized a set of surface markers used in mucosal immunity research to investigate the involvement of macrophages and DC subpopulations in post-stroke inflammation in mice.

Findings: Photothrombotic stroke induced a significant increase of lineage (CD3, B220, Ly6G and CD49b) negative CD11b+ cells in the brain primarily consisting of F4/80+ macrophages and, to a lesser extent, F4/80-/CD11c-/CD11b+ monocytes and F4/80-/CD11c+ DC. The latter could be differentiated into the classical migratory DC subpopulations (CD11b+ and CD103+), but no CD4 or CD8+ DC were found. Finally, stroke caused a significant increase of CD11b/CD103 double-positive DC in the affected brain hemisphere.

Conclusions: The surface marker combination used in this study allowed a phenotypic differentiation of macrophages and DC subpopulations after stroke, thus providing an important prerequisite to study post-stroke immunity and tolerance.

No MeSH data available.


Related in: MedlinePlus

Subdifferentiation of Lin-/CD11c+ cells. (A), in the ipsilateral (ipsi) hemisphere, Lin-/CD11c+ cells were primarily composed of CD11b+/CD103- and CD11b+/CD103+ cells. We found only few CD11b-/CD103+ and CD11b-/CD103- cells, but no CD4+ or CD8+ expression. (B), Lin-/CD11c+ cells were predominantly MHCII+. CD11b+/CD103+ DC were enriched in the MHCII+ population (green), but also present among MHCII- cells (black; experiment 2, n = 3). *p < 0.05, **p < 0.01 by one way ANOVA and Bonferronis’ post-hoc test. Contra, contralateral.
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Fig3: Subdifferentiation of Lin-/CD11c+ cells. (A), in the ipsilateral (ipsi) hemisphere, Lin-/CD11c+ cells were primarily composed of CD11b+/CD103- and CD11b+/CD103+ cells. We found only few CD11b-/CD103+ and CD11b-/CD103- cells, but no CD4+ or CD8+ expression. (B), Lin-/CD11c+ cells were predominantly MHCII+. CD11b+/CD103+ DC were enriched in the MHCII+ population (green), but also present among MHCII- cells (black; experiment 2, n = 3). *p < 0.05, **p < 0.01 by one way ANOVA and Bonferronis’ post-hoc test. Contra, contralateral.

Mentions: Next, we differentiated Lin-/CD11c+ cDC by CD11b and CD103 as classical markers for non-lymphoid tissue DCs. Two major subpopulations responded to ischemic stroke: a CD11b+/CD103- population and CD11b+/CD103+ cells (Figure 3A) that may resemble migratory and tolerogenic DCs in the intestinal lamina propria [14]. CD11b+/CD103+ cells were virtually absent in naive brain tissue, but 30fold increased in the ischemic hemisphere. Neither CD11b-/CD103- cells (Figure 3A) nor CD11b+/CD103- and CD11b-/CD103+ cells (not shown) expressed CD4 or CD8.Figure 3


Flow cytometric characterization of brain dendritic cell subsets after murine stroke.

Pösel C, Uri A, Schulz I, Boltze J, Weise G, Wagner DC - Exp Transl Stroke Med (2014)

Subdifferentiation of Lin-/CD11c+ cells. (A), in the ipsilateral (ipsi) hemisphere, Lin-/CD11c+ cells were primarily composed of CD11b+/CD103- and CD11b+/CD103+ cells. We found only few CD11b-/CD103+ and CD11b-/CD103- cells, but no CD4+ or CD8+ expression. (B), Lin-/CD11c+ cells were predominantly MHCII+. CD11b+/CD103+ DC were enriched in the MHCII+ population (green), but also present among MHCII- cells (black; experiment 2, n = 3). *p < 0.05, **p < 0.01 by one way ANOVA and Bonferronis’ post-hoc test. Contra, contralateral.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230520&req=5

Fig3: Subdifferentiation of Lin-/CD11c+ cells. (A), in the ipsilateral (ipsi) hemisphere, Lin-/CD11c+ cells were primarily composed of CD11b+/CD103- and CD11b+/CD103+ cells. We found only few CD11b-/CD103+ and CD11b-/CD103- cells, but no CD4+ or CD8+ expression. (B), Lin-/CD11c+ cells were predominantly MHCII+. CD11b+/CD103+ DC were enriched in the MHCII+ population (green), but also present among MHCII- cells (black; experiment 2, n = 3). *p < 0.05, **p < 0.01 by one way ANOVA and Bonferronis’ post-hoc test. Contra, contralateral.
Mentions: Next, we differentiated Lin-/CD11c+ cDC by CD11b and CD103 as classical markers for non-lymphoid tissue DCs. Two major subpopulations responded to ischemic stroke: a CD11b+/CD103- population and CD11b+/CD103+ cells (Figure 3A) that may resemble migratory and tolerogenic DCs in the intestinal lamina propria [14]. CD11b+/CD103+ cells were virtually absent in naive brain tissue, but 30fold increased in the ischemic hemisphere. Neither CD11b-/CD103- cells (Figure 3A) nor CD11b+/CD103- and CD11b-/CD103+ cells (not shown) expressed CD4 or CD8.Figure 3

Bottom Line: Sterile inflammation is a substantial element of post-stroke pathophysiology with the determination of autoimmunity versus tolerance being one of its most important aspects.The latter could be differentiated into the classical migratory DC subpopulations (CD11b+ and CD103+), but no CD4 or CD8+ DC were found.Finally, stroke caused a significant increase of CD11b/CD103 double-positive DC in the affected brain hemisphere.

View Article: PubMed Central - PubMed

Affiliation: Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.

ABSTRACT

Background: Sterile inflammation is a substantial element of post-stroke pathophysiology with the determination of autoimmunity versus tolerance being one of its most important aspects. It is believed that this determination is initiated relatively early after stroke onset by clearing macrophages and migratory dendritic cells (DC). However, the phenotypic differentiation of macrophages and DC is intricate particularly in the disease context. Here, we utilized a set of surface markers used in mucosal immunity research to investigate the involvement of macrophages and DC subpopulations in post-stroke inflammation in mice.

Findings: Photothrombotic stroke induced a significant increase of lineage (CD3, B220, Ly6G and CD49b) negative CD11b+ cells in the brain primarily consisting of F4/80+ macrophages and, to a lesser extent, F4/80-/CD11c-/CD11b+ monocytes and F4/80-/CD11c+ DC. The latter could be differentiated into the classical migratory DC subpopulations (CD11b+ and CD103+), but no CD4 or CD8+ DC were found. Finally, stroke caused a significant increase of CD11b/CD103 double-positive DC in the affected brain hemisphere.

Conclusions: The surface marker combination used in this study allowed a phenotypic differentiation of macrophages and DC subpopulations after stroke, thus providing an important prerequisite to study post-stroke immunity and tolerance.

No MeSH data available.


Related in: MedlinePlus