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Cell therapy centered on IL-1Ra is neuroprotective in experimental stroke.

Clausen BH, Lambertsen KL, Dagnæs-Hansen F, Babcock AA, von Linstow CU, Meldgaard M, Kristensen BW, Deierborg T, Finsen B - Acta Neuropathol. (2016)

Bottom Line: The IL-1Ra-producing bone marrow cells increase the number of IL-1Ra-producing microglia, reduce the availability of IL-1β, and modulate mitogen-activated protein kinase (MAPK) signaling in the ischemic cortex.The importance of these results is underlined by demonstration of IL-1Ra-producing cells in the human cortex early after ischemic stroke.Taken together, our results attribute distinct neuroprotective or neurotoxic functions to segregated subsets of microglia and suggest that treatment strategies increasing the production of IL-1Ra by infiltrating leukocytes or microglia may also be neuroprotective if applied early after stroke onset in patients.

View Article: PubMed Central - PubMed

Affiliation: Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, J. B. Winsloewsvej 25, 5000, Odense C, Denmark. bclausen@health.sdu.dk.

ABSTRACT
Cell-based therapies are emerging as new promising treatments in stroke. However, their functional mechanism and therapeutic potential during early infarct maturation has so far received little attention. Here, we asked if cell-based delivery of the interleukin-1 receptor antagonist (IL-1Ra), a known neuroprotectant in stroke, can promote neuroprotection, by modulating the detrimental inflammatory response in the tissue at risk. We show by the use of IL-1Ra-overexpressing and IL-1Ra-deficient mice that IL-1Ra is neuroprotective in stroke. Characterization of the cellular and spatiotemporal production of IL-1Ra and IL-1α/β identifies microglia, not infiltrating leukocytes, as the major sources of IL-1Ra after experimental stroke, and shows IL-1Ra and IL-1β to be produced by segregated subsets of microglia with a small proportion of these cells co-expressing IL-1α. Reconstitution of whole body irradiated mice with IL-1Ra-producing bone marrow cells is associated with neuroprotection and recruitment of IL-1Ra-producing leukocytes after stroke. Neuroprotection is also achieved by therapeutic injection of IL-1Ra-producing bone marrow cells 30 min after stroke onset, additionally improving the functional outcome in two different stroke models. The IL-1Ra-producing bone marrow cells increase the number of IL-1Ra-producing microglia, reduce the availability of IL-1β, and modulate mitogen-activated protein kinase (MAPK) signaling in the ischemic cortex. The importance of these results is underlined by demonstration of IL-1Ra-producing cells in the human cortex early after ischemic stroke. Taken together, our results attribute distinct neuroprotective or neurotoxic functions to segregated subsets of microglia and suggest that treatment strategies increasing the production of IL-1Ra by infiltrating leukocytes or microglia may also be neuroprotective if applied early after stroke onset in patients.

No MeSH data available.


Related in: MedlinePlus

IL-1Ra and IL-1α/β synthesis is increased in CD11b+ cells in the peri-infarct after pMCAo. IL-1Ra (a–g), IL-1α (h–m), and IL-1β (n–s) mRNA and protein expression, and IHC double-fluorescence stainings (t–x) after pMCAo. a, h, n Quantitative PCR results showing temporal changes in IL-1Ra, IL-1α and IL-1β mRNA levels in pMCAo- and sham-operated mice, compared to non-lesioned controls (Ctl), n = 10–12/group. Statistical data are presented as mean ± SD (Kruskal–Wallis test with Dunns post hoc test). *P < 0.05; **P < 0.01; ***P < 0.001. b–d, i–k, o–qISH showing IL-1Ra mRNA+ (b, c, d), IL-1α mRNA+ (i, j, k) and IL-1β mRNA+ (o, p, q) cells in the peri-infarct. By 24 h, IL-1β mRNA was expressed by single cells and vessel-associated cells (q). e–g, l, m, r, s IHC showing IL-1Ra+ cells (e–g), IL-1α+ cells and (l, m), IL-1β+ (r, s) cells in the peri-infarct. t–v IHC double-fluorescence staining showing co-localization of IL-1Ra (t), IL-1α (u) and IL-1β (v) to CD11b+ microglia 24 h after pMCAo. w, x IHC double-fluorescence staining showing co-localization of IL-1α to CD41+ platelets in the peri-infarct, 4 and 6 h (w), but not 24 h after pMCAo (x). Scale bars: 20 µm (b–d, i–k, o–q), 10 µm (e, f, g, l, m, r–x , and insert in g) and 100 µm (g)
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Fig2: IL-1Ra and IL-1α/β synthesis is increased in CD11b+ cells in the peri-infarct after pMCAo. IL-1Ra (a–g), IL-1α (h–m), and IL-1β (n–s) mRNA and protein expression, and IHC double-fluorescence stainings (t–x) after pMCAo. a, h, n Quantitative PCR results showing temporal changes in IL-1Ra, IL-1α and IL-1β mRNA levels in pMCAo- and sham-operated mice, compared to non-lesioned controls (Ctl), n = 10–12/group. Statistical data are presented as mean ± SD (Kruskal–Wallis test with Dunns post hoc test). *P < 0.05; **P < 0.01; ***P < 0.001. b–d, i–k, o–qISH showing IL-1Ra mRNA+ (b, c, d), IL-1α mRNA+ (i, j, k) and IL-1β mRNA+ (o, p, q) cells in the peri-infarct. By 24 h, IL-1β mRNA was expressed by single cells and vessel-associated cells (q). e–g, l, m, r, s IHC showing IL-1Ra+ cells (e–g), IL-1α+ cells and (l, m), IL-1β+ (r, s) cells in the peri-infarct. t–v IHC double-fluorescence staining showing co-localization of IL-1Ra (t), IL-1α (u) and IL-1β (v) to CD11b+ microglia 24 h after pMCAo. w, x IHC double-fluorescence staining showing co-localization of IL-1α to CD41+ platelets in the peri-infarct, 4 and 6 h (w), but not 24 h after pMCAo (x). Scale bars: 20 µm (b–d, i–k, o–q), 10 µm (e, f, g, l, m, r–x , and insert in g) and 100 µm (g)

Mentions: To obtain detailed insight into the cellular orchestration of IL-1Ra and IL-1(α/β), we examined the synthesis of all three cytokines in C57BL/6 mice after pMCAo. By the use of quantitative reverse transcription PCR (qPCR), we observed small fluctuations in IL-1Ra mRNA until 6 h (Fig. 2a) and a significant up-regulation at 12 and 24 h after pMCAo, compared to non-lesioned and sham controls (Fig. 2a). Within the peri-infarct, we found multiple process-bearing microglial-like cells with abundant IL-1Ra mRNA (Fig. 2b–d) and protein accumulation (Fig. 2f, g) 6, 12 and 24 h after pMCAo, and round vessel-associated leukocyte-like cells from 4 to 6 h after pMCAo (insert in Fig. 2b, e).Fig. 2


Cell therapy centered on IL-1Ra is neuroprotective in experimental stroke.

Clausen BH, Lambertsen KL, Dagnæs-Hansen F, Babcock AA, von Linstow CU, Meldgaard M, Kristensen BW, Deierborg T, Finsen B - Acta Neuropathol. (2016)

IL-1Ra and IL-1α/β synthesis is increased in CD11b+ cells in the peri-infarct after pMCAo. IL-1Ra (a–g), IL-1α (h–m), and IL-1β (n–s) mRNA and protein expression, and IHC double-fluorescence stainings (t–x) after pMCAo. a, h, n Quantitative PCR results showing temporal changes in IL-1Ra, IL-1α and IL-1β mRNA levels in pMCAo- and sham-operated mice, compared to non-lesioned controls (Ctl), n = 10–12/group. Statistical data are presented as mean ± SD (Kruskal–Wallis test with Dunns post hoc test). *P < 0.05; **P < 0.01; ***P < 0.001. b–d, i–k, o–qISH showing IL-1Ra mRNA+ (b, c, d), IL-1α mRNA+ (i, j, k) and IL-1β mRNA+ (o, p, q) cells in the peri-infarct. By 24 h, IL-1β mRNA was expressed by single cells and vessel-associated cells (q). e–g, l, m, r, s IHC showing IL-1Ra+ cells (e–g), IL-1α+ cells and (l, m), IL-1β+ (r, s) cells in the peri-infarct. t–v IHC double-fluorescence staining showing co-localization of IL-1Ra (t), IL-1α (u) and IL-1β (v) to CD11b+ microglia 24 h after pMCAo. w, x IHC double-fluorescence staining showing co-localization of IL-1α to CD41+ platelets in the peri-infarct, 4 and 6 h (w), but not 24 h after pMCAo (x). Scale bars: 20 µm (b–d, i–k, o–q), 10 µm (e, f, g, l, m, r–x , and insert in g) and 100 µm (g)
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Fig2: IL-1Ra and IL-1α/β synthesis is increased in CD11b+ cells in the peri-infarct after pMCAo. IL-1Ra (a–g), IL-1α (h–m), and IL-1β (n–s) mRNA and protein expression, and IHC double-fluorescence stainings (t–x) after pMCAo. a, h, n Quantitative PCR results showing temporal changes in IL-1Ra, IL-1α and IL-1β mRNA levels in pMCAo- and sham-operated mice, compared to non-lesioned controls (Ctl), n = 10–12/group. Statistical data are presented as mean ± SD (Kruskal–Wallis test with Dunns post hoc test). *P < 0.05; **P < 0.01; ***P < 0.001. b–d, i–k, o–qISH showing IL-1Ra mRNA+ (b, c, d), IL-1α mRNA+ (i, j, k) and IL-1β mRNA+ (o, p, q) cells in the peri-infarct. By 24 h, IL-1β mRNA was expressed by single cells and vessel-associated cells (q). e–g, l, m, r, s IHC showing IL-1Ra+ cells (e–g), IL-1α+ cells and (l, m), IL-1β+ (r, s) cells in the peri-infarct. t–v IHC double-fluorescence staining showing co-localization of IL-1Ra (t), IL-1α (u) and IL-1β (v) to CD11b+ microglia 24 h after pMCAo. w, x IHC double-fluorescence staining showing co-localization of IL-1α to CD41+ platelets in the peri-infarct, 4 and 6 h (w), but not 24 h after pMCAo (x). Scale bars: 20 µm (b–d, i–k, o–q), 10 µm (e, f, g, l, m, r–x , and insert in g) and 100 µm (g)
Mentions: To obtain detailed insight into the cellular orchestration of IL-1Ra and IL-1(α/β), we examined the synthesis of all three cytokines in C57BL/6 mice after pMCAo. By the use of quantitative reverse transcription PCR (qPCR), we observed small fluctuations in IL-1Ra mRNA until 6 h (Fig. 2a) and a significant up-regulation at 12 and 24 h after pMCAo, compared to non-lesioned and sham controls (Fig. 2a). Within the peri-infarct, we found multiple process-bearing microglial-like cells with abundant IL-1Ra mRNA (Fig. 2b–d) and protein accumulation (Fig. 2f, g) 6, 12 and 24 h after pMCAo, and round vessel-associated leukocyte-like cells from 4 to 6 h after pMCAo (insert in Fig. 2b, e).Fig. 2

Bottom Line: The IL-1Ra-producing bone marrow cells increase the number of IL-1Ra-producing microglia, reduce the availability of IL-1β, and modulate mitogen-activated protein kinase (MAPK) signaling in the ischemic cortex.The importance of these results is underlined by demonstration of IL-1Ra-producing cells in the human cortex early after ischemic stroke.Taken together, our results attribute distinct neuroprotective or neurotoxic functions to segregated subsets of microglia and suggest that treatment strategies increasing the production of IL-1Ra by infiltrating leukocytes or microglia may also be neuroprotective if applied early after stroke onset in patients.

View Article: PubMed Central - PubMed

Affiliation: Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, J. B. Winsloewsvej 25, 5000, Odense C, Denmark. bclausen@health.sdu.dk.

ABSTRACT
Cell-based therapies are emerging as new promising treatments in stroke. However, their functional mechanism and therapeutic potential during early infarct maturation has so far received little attention. Here, we asked if cell-based delivery of the interleukin-1 receptor antagonist (IL-1Ra), a known neuroprotectant in stroke, can promote neuroprotection, by modulating the detrimental inflammatory response in the tissue at risk. We show by the use of IL-1Ra-overexpressing and IL-1Ra-deficient mice that IL-1Ra is neuroprotective in stroke. Characterization of the cellular and spatiotemporal production of IL-1Ra and IL-1α/β identifies microglia, not infiltrating leukocytes, as the major sources of IL-1Ra after experimental stroke, and shows IL-1Ra and IL-1β to be produced by segregated subsets of microglia with a small proportion of these cells co-expressing IL-1α. Reconstitution of whole body irradiated mice with IL-1Ra-producing bone marrow cells is associated with neuroprotection and recruitment of IL-1Ra-producing leukocytes after stroke. Neuroprotection is also achieved by therapeutic injection of IL-1Ra-producing bone marrow cells 30 min after stroke onset, additionally improving the functional outcome in two different stroke models. The IL-1Ra-producing bone marrow cells increase the number of IL-1Ra-producing microglia, reduce the availability of IL-1β, and modulate mitogen-activated protein kinase (MAPK) signaling in the ischemic cortex. The importance of these results is underlined by demonstration of IL-1Ra-producing cells in the human cortex early after ischemic stroke. Taken together, our results attribute distinct neuroprotective or neurotoxic functions to segregated subsets of microglia and suggest that treatment strategies increasing the production of IL-1Ra by infiltrating leukocytes or microglia may also be neuroprotective if applied early after stroke onset in patients.

No MeSH data available.


Related in: MedlinePlus