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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-overproducing BM cells stimulate microglial production of IL-1Ra. a GFP+ BM cells in the cortex of C57BL/6 mice 2, 4 and 6 h after pMCAo, but not in non-lesioned controls (Ctl). b BM profiles, cells numbers and IL-1Ra mRNA expression in BM from IL-1Ra-Tg (Tg) and LM mice. cDot plots showing flow cytometric profiles of CFSE+ BM cells 6 h after pMCAo (5.5 h after BM transplantation). d Infiltrating CFSE+ BM cells and CFSE− CD11b+CD45high leukocytes in LM–LM and Tg–LM mice 6 h after pMCAo. e MFI of IL-1Ra in gated CFSE+ BM cells 6 h after pMCAo, the CFSE+ BM cells originated from LM and IL-1Ra-Tg (Tg) donor mice. f–iDot plots showing flow cytometric profiles with isotype quadrants (f, h) and quantification of IL-1Ra+ CD11b+CD45highCFSE− leukocytes (g) and IL-1Ra+ CD11b+CD45dimCFSE− microglia (i) in LM, LM–LM and Tg–LM 6 h after pMCAo. Statistical data are presented as mean ± SD, n = 4/group (Kruskal–Wallis test with Dunns post hoc test), *#P < 0.05, **##P < 0.01. Scale bar 100 µm (a) and 10 µm (c)
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Fig5: IL-1Ra-overproducing BM cells stimulate microglial production of IL-1Ra. a GFP+ BM cells in the cortex of C57BL/6 mice 2, 4 and 6 h after pMCAo, but not in non-lesioned controls (Ctl). b BM profiles, cells numbers and IL-1Ra mRNA expression in BM from IL-1Ra-Tg (Tg) and LM mice. cDot plots showing flow cytometric profiles of CFSE+ BM cells 6 h after pMCAo (5.5 h after BM transplantation). d Infiltrating CFSE+ BM cells and CFSE− CD11b+CD45high leukocytes in LM–LM and Tg–LM mice 6 h after pMCAo. e MFI of IL-1Ra in gated CFSE+ BM cells 6 h after pMCAo, the CFSE+ BM cells originated from LM and IL-1Ra-Tg (Tg) donor mice. f–iDot plots showing flow cytometric profiles with isotype quadrants (f, h) and quantification of IL-1Ra+ CD11b+CD45highCFSE− leukocytes (g) and IL-1Ra+ CD11b+CD45dimCFSE− microglia (i) in LM, LM–LM and Tg–LM 6 h after pMCAo. Statistical data are presented as mean ± SD, n = 4/group (Kruskal–Wallis test with Dunns post hoc test), *#P < 0.05, **##P < 0.01. Scale bar 100 µm (a) and 10 µm (c)

Mentions: Having shown that leukocyte-derived IL-1Ra can be neuroprotective using irradiation BM chimeric mice, we next tested the ability of BM cells to infiltrate the ischemic cortex when injected post-stroke. First, we evaluated the ability of BM cells to infiltrate the developing infarct, by injecting GFP+ BM cells into the tail vein of C57BL/6 mice 30 min after pMCAo. We identified GFP+ cells as soon as 2 h after pMCAo, increasing in numbers at 4 and 6 h (n = 4/group), while no cells were found in the cortex of non-lesioned controls (Fig. 5a). Next, we analyzed BM samples isolated from IL-1Ra-Tg and LM mice, and showed that samples, as expected, were similar regarding the BM profiles and the unit volume (cells/ml), but that BM cells from IL-1Ra-Tg mice showed higher expression of IL-1Ra mRNA than BM cells from sibling donors (Fig. 5b).Fig. 5


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-overproducing BM cells stimulate microglial production of IL-1Ra. a GFP+ BM cells in the cortex of C57BL/6 mice 2, 4 and 6 h after pMCAo, but not in non-lesioned controls (Ctl). b BM profiles, cells numbers and IL-1Ra mRNA expression in BM from IL-1Ra-Tg (Tg) and LM mice. cDot plots showing flow cytometric profiles of CFSE+ BM cells 6 h after pMCAo (5.5 h after BM transplantation). d Infiltrating CFSE+ BM cells and CFSE− CD11b+CD45high leukocytes in LM–LM and Tg–LM mice 6 h after pMCAo. e MFI of IL-1Ra in gated CFSE+ BM cells 6 h after pMCAo, the CFSE+ BM cells originated from LM and IL-1Ra-Tg (Tg) donor mice. f–iDot plots showing flow cytometric profiles with isotype quadrants (f, h) and quantification of IL-1Ra+ CD11b+CD45highCFSE− leukocytes (g) and IL-1Ra+ CD11b+CD45dimCFSE− microglia (i) in LM, LM–LM and Tg–LM 6 h after pMCAo. Statistical data are presented as mean ± SD, n = 4/group (Kruskal–Wallis test with Dunns post hoc test), *#P < 0.05, **##P < 0.01. Scale bar 100 µm (a) and 10 µm (c)
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Fig5: IL-1Ra-overproducing BM cells stimulate microglial production of IL-1Ra. a GFP+ BM cells in the cortex of C57BL/6 mice 2, 4 and 6 h after pMCAo, but not in non-lesioned controls (Ctl). b BM profiles, cells numbers and IL-1Ra mRNA expression in BM from IL-1Ra-Tg (Tg) and LM mice. cDot plots showing flow cytometric profiles of CFSE+ BM cells 6 h after pMCAo (5.5 h after BM transplantation). d Infiltrating CFSE+ BM cells and CFSE− CD11b+CD45high leukocytes in LM–LM and Tg–LM mice 6 h after pMCAo. e MFI of IL-1Ra in gated CFSE+ BM cells 6 h after pMCAo, the CFSE+ BM cells originated from LM and IL-1Ra-Tg (Tg) donor mice. f–iDot plots showing flow cytometric profiles with isotype quadrants (f, h) and quantification of IL-1Ra+ CD11b+CD45highCFSE− leukocytes (g) and IL-1Ra+ CD11b+CD45dimCFSE− microglia (i) in LM, LM–LM and Tg–LM 6 h after pMCAo. Statistical data are presented as mean ± SD, n = 4/group (Kruskal–Wallis test with Dunns post hoc test), *#P < 0.05, **##P < 0.01. Scale bar 100 µm (a) and 10 µm (c)
Mentions: Having shown that leukocyte-derived IL-1Ra can be neuroprotective using irradiation BM chimeric mice, we next tested the ability of BM cells to infiltrate the ischemic cortex when injected post-stroke. First, we evaluated the ability of BM cells to infiltrate the developing infarct, by injecting GFP+ BM cells into the tail vein of C57BL/6 mice 30 min after pMCAo. We identified GFP+ cells as soon as 2 h after pMCAo, increasing in numbers at 4 and 6 h (n = 4/group), while no cells were found in the cortex of non-lesioned controls (Fig. 5a). Next, we analyzed BM samples isolated from IL-1Ra-Tg and LM mice, and showed that samples, as expected, were similar regarding the BM profiles and the unit volume (cells/ml), but that BM cells from IL-1Ra-Tg mice showed higher expression of IL-1Ra mRNA than BM cells from sibling donors (Fig. 5b).Fig. 5

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