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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 have a therapeutic effect after both pMCAo and tMCAo. a–c Toluidine blue stainings and infarct volume estimation 24 h after pMCAo, n = 14/group (a), 5 days after pMCAo, n = 14/group (b), and 24 h after tMCAo, n = 12–17/group (c), with infarct volume estimation shown for LM, LM–LM and Tg–LM mice. d–f Quantitative PCR and ELISA performed on sections parallel to those used in (a–c) show differences in IL-1β mRNA and protein among the treatment groups. g, h Grip strength of the affected right front paws (g) and grip strength asymmetry in right versus left front paw (h) in LM, LM–LM and Tg–LM mice 1, 2 and 5 days after pMCAo compared to baseline. The stippled line (g) indicates the grip strength of the left front paw. i Grip strength asymmetry in right versus left front paw in LM, LM–LM and Tg–LM mice 1 day after tMCAo. Statistical data are presented as mean ± SD. (Mann–Whitney test (a, d), Kruskal–Wallis test with Dunns post hoc test (b, c, e, f), repeated-measures ANOVA with Dunnett post hoc test (g) or Wilcoxon matched pairs test (h, i). *P < 0.05; **P < 0.01. B baseline. Scale bars 1 mm
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Fig6: IL-1Ra-overproducing BM cells have a therapeutic effect after both pMCAo and tMCAo. a–c Toluidine blue stainings and infarct volume estimation 24 h after pMCAo, n = 14/group (a), 5 days after pMCAo, n = 14/group (b), and 24 h after tMCAo, n = 12–17/group (c), with infarct volume estimation shown for LM, LM–LM and Tg–LM mice. d–f Quantitative PCR and ELISA performed on sections parallel to those used in (a–c) show differences in IL-1β mRNA and protein among the treatment groups. g, h Grip strength of the affected right front paws (g) and grip strength asymmetry in right versus left front paw (h) in LM, LM–LM and Tg–LM mice 1, 2 and 5 days after pMCAo compared to baseline. The stippled line (g) indicates the grip strength of the left front paw. i Grip strength asymmetry in right versus left front paw in LM, LM–LM and Tg–LM mice 1 day after tMCAo. Statistical data are presented as mean ± SD. (Mann–Whitney test (a, d), Kruskal–Wallis test with Dunns post hoc test (b, c, e, f), repeated-measures ANOVA with Dunnett post hoc test (g) or Wilcoxon matched pairs test (h, i). *P < 0.05; **P < 0.01. B baseline. Scale bars 1 mm

Mentions: As an increase in IL-1Ra production by BM cells or microglia could impact the infarct development, thereby changing the neurological outcome, we finally investigated the therapeutic effect of IL-1Ra-overproducing BM cells in mice subjected to either pMCAo or tMCAo (Fig. 6). BM cells harvested from either LM mice or IL-1Ra-Tg mice (Tg) were injected into the tail veins of LM recipients 30 min after MCAo (Table S1). We first investigated the effect of IL-1Ra-overproducing BM cells in mice subjected to pMCAo with 24 h or 5 days survival (Table S1). We observed a significant reduction in infarct size in Tg–LM compared to LM–LM mice 24 h after pMCAo (survival rate/group >95 %) (Fig. 6a). LM–LM mice showed similar infarct sizes as non-treated LM mice 24 h after pMCAo (compared to Fig. 1b). Encouraged by the prominent effect of IL-1Ra+ BM cells, we allowed mice to survive for 5 days to assess any physiological and/or behavioral improvements of this treatment strategy. By day 5, we observed a significant reduction in infarct volume in Tg–LM mice compared to LM–LM mice (survival rate/group >95 %), with no difference between LM–LM-treated mice and LM mice (Fig. 6b). The protective effect of IL-1Ra was not a result of decreased body temperature, as rectal temperature recordings (up to 3 h) showed no differences between treatment groups (Figure S5a). Additionally, blood gas parameters (pO2/pCO2, pH, electrolytes, glucose/lactate) recorded 30 min after BM injections (i.e., 1 h after pMCAo) showed no difference among various experimental groups (LM, LM–LM and Tg–LM mice) or when compared to non-lesioned mice (Figure S5b-i). When performing in situ hybridization, we observed IL-1Ra mRNA+ cells in the peri-infarct in none of the ten LM mice, one out of ten LM–LM and in two out of ten Tg–LM mice 5 days after pMCAo (Figure S6a, shown for Tg–LM). These results matched qPCR results obtained on RNA isolated from parallel sections, showing elevated levels of IL-1Ra mRNA in Tg–LM compared to both LM–LM and LM 5 days after pMCAo (Figure S6b). These results suggest the continued presence of infiltrating IL-1Ra mRNA+ BM cells, and/or a prolonged expression of IL-1Ra mRNA by host cells in BM-grafted mice, 5 days after pMCAo.Fig. 6


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 have a therapeutic effect after both pMCAo and tMCAo. a–c Toluidine blue stainings and infarct volume estimation 24 h after pMCAo, n = 14/group (a), 5 days after pMCAo, n = 14/group (b), and 24 h after tMCAo, n = 12–17/group (c), with infarct volume estimation shown for LM, LM–LM and Tg–LM mice. d–f Quantitative PCR and ELISA performed on sections parallel to those used in (a–c) show differences in IL-1β mRNA and protein among the treatment groups. g, h Grip strength of the affected right front paws (g) and grip strength asymmetry in right versus left front paw (h) in LM, LM–LM and Tg–LM mice 1, 2 and 5 days after pMCAo compared to baseline. The stippled line (g) indicates the grip strength of the left front paw. i Grip strength asymmetry in right versus left front paw in LM, LM–LM and Tg–LM mice 1 day after tMCAo. Statistical data are presented as mean ± SD. (Mann–Whitney test (a, d), Kruskal–Wallis test with Dunns post hoc test (b, c, e, f), repeated-measures ANOVA with Dunnett post hoc test (g) or Wilcoxon matched pairs test (h, i). *P < 0.05; **P < 0.01. B baseline. Scale bars 1 mm
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Fig6: IL-1Ra-overproducing BM cells have a therapeutic effect after both pMCAo and tMCAo. a–c Toluidine blue stainings and infarct volume estimation 24 h after pMCAo, n = 14/group (a), 5 days after pMCAo, n = 14/group (b), and 24 h after tMCAo, n = 12–17/group (c), with infarct volume estimation shown for LM, LM–LM and Tg–LM mice. d–f Quantitative PCR and ELISA performed on sections parallel to those used in (a–c) show differences in IL-1β mRNA and protein among the treatment groups. g, h Grip strength of the affected right front paws (g) and grip strength asymmetry in right versus left front paw (h) in LM, LM–LM and Tg–LM mice 1, 2 and 5 days after pMCAo compared to baseline. The stippled line (g) indicates the grip strength of the left front paw. i Grip strength asymmetry in right versus left front paw in LM, LM–LM and Tg–LM mice 1 day after tMCAo. Statistical data are presented as mean ± SD. (Mann–Whitney test (a, d), Kruskal–Wallis test with Dunns post hoc test (b, c, e, f), repeated-measures ANOVA with Dunnett post hoc test (g) or Wilcoxon matched pairs test (h, i). *P < 0.05; **P < 0.01. B baseline. Scale bars 1 mm
Mentions: As an increase in IL-1Ra production by BM cells or microglia could impact the infarct development, thereby changing the neurological outcome, we finally investigated the therapeutic effect of IL-1Ra-overproducing BM cells in mice subjected to either pMCAo or tMCAo (Fig. 6). BM cells harvested from either LM mice or IL-1Ra-Tg mice (Tg) were injected into the tail veins of LM recipients 30 min after MCAo (Table S1). We first investigated the effect of IL-1Ra-overproducing BM cells in mice subjected to pMCAo with 24 h or 5 days survival (Table S1). We observed a significant reduction in infarct size in Tg–LM compared to LM–LM mice 24 h after pMCAo (survival rate/group >95 %) (Fig. 6a). LM–LM mice showed similar infarct sizes as non-treated LM mice 24 h after pMCAo (compared to Fig. 1b). Encouraged by the prominent effect of IL-1Ra+ BM cells, we allowed mice to survive for 5 days to assess any physiological and/or behavioral improvements of this treatment strategy. By day 5, we observed a significant reduction in infarct volume in Tg–LM mice compared to LM–LM mice (survival rate/group >95 %), with no difference between LM–LM-treated mice and LM mice (Fig. 6b). The protective effect of IL-1Ra was not a result of decreased body temperature, as rectal temperature recordings (up to 3 h) showed no differences between treatment groups (Figure S5a). Additionally, blood gas parameters (pO2/pCO2, pH, electrolytes, glucose/lactate) recorded 30 min after BM injections (i.e., 1 h after pMCAo) showed no difference among various experimental groups (LM, LM–LM and Tg–LM mice) or when compared to non-lesioned mice (Figure S5b-i). When performing in situ hybridization, we observed IL-1Ra mRNA+ cells in the peri-infarct in none of the ten LM mice, one out of ten LM–LM and in two out of ten Tg–LM mice 5 days after pMCAo (Figure S6a, shown for Tg–LM). These results matched qPCR results obtained on RNA isolated from parallel sections, showing elevated levels of IL-1Ra mRNA in Tg–LM compared to both LM–LM and LM 5 days after pMCAo (Figure S6b). These results suggest the continued presence of infiltrating IL-1Ra mRNA+ BM cells, and/or a prolonged expression of IL-1Ra mRNA by host cells in BM-grafted mice, 5 days after pMCAo.Fig. 6

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