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Characterisation of immune cell function in fragment and full-length Huntington's disease mouse models.

Träger U, Andre R, Magnusson-Lind A, Miller JR, Connolly C, Weiss A, Grueninger S, Silajdžić E, Smith DL, Leavitt BR, Bates GP, Björkqvist M, Tabrizi SJ - Neurobiol. Dis. (2014)

Bottom Line: Inflammation is a growing area of research in neurodegeneration.However, bone marrow CD11b(+) cells did not show the same hyper-responsive phenotype as spleen and blood cells.Taken together, these results show significant promise for these mouse models to be used to study targeting innate immune pathways identified in human cells, thereby helping to understand the role the peripheral immune system plays in HD progression.

View Article: PubMed Central - PubMed

Affiliation: UCL Institute of Neurology, Dept. of Neurodegenerative Disease, London, UK.

No MeSH data available.


Related in: MedlinePlus

Altered cytokine production by myeloid cells from R6/2 mice. (A) CD11b+ myeloid cells were isolated from pooled blood samples obtained from 12-week old R6/2 or wild-type mice. Cell cultures primed with IFNγ and stimulated with LPS for 24 h showed significant differences in IL-6 and TNFα production. (B) R6/2 spleen and (C) R6/2 bone marrow myeloid cells were isolated using anti-CD11b magnetic beads, and individual cultures from each mouse were stimulated with IFNγ and LPS for 24 h. Measuring cytokine production in supernatants by multiplex ELISA, R6/2 spleen myeloid cells demonstrated changes in IL-1β levels, whilst bone marrow myeloid cells from R6/2 mice showed the same cytokine levels as wild-type cells. Graphs show mean concentrations corrected to protein content ± SEM. Unpaired two-tailed Student's t-tests. (A) n = technical replicates of pooled samples from different mice, (B + C) n = biological replicates representing individual mice.
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f0005: Altered cytokine production by myeloid cells from R6/2 mice. (A) CD11b+ myeloid cells were isolated from pooled blood samples obtained from 12-week old R6/2 or wild-type mice. Cell cultures primed with IFNγ and stimulated with LPS for 24 h showed significant differences in IL-6 and TNFα production. (B) R6/2 spleen and (C) R6/2 bone marrow myeloid cells were isolated using anti-CD11b magnetic beads, and individual cultures from each mouse were stimulated with IFNγ and LPS for 24 h. Measuring cytokine production in supernatants by multiplex ELISA, R6/2 spleen myeloid cells demonstrated changes in IL-1β levels, whilst bone marrow myeloid cells from R6/2 mice showed the same cytokine levels as wild-type cells. Graphs show mean concentrations corrected to protein content ± SEM. Unpaired two-tailed Student's t-tests. (A) n = technical replicates of pooled samples from different mice, (B + C) n = biological replicates representing individual mice.

Mentions: To determine whether monocytes from HD mouse models are hyper-reactive like those from HD patient blood (Träger et al., 2014), CD11b+ cells from 12-week old R6/2 mice were isolated using magnetic cell sorting and seeded in culture, then primed with IFNγ and stimulated with LPS for 24 h. Analysis of culture supernatants using a multiplex ELISA platform showed an increase in IL-6 and TNFα levels, but not IL-1β, IL-10 and IL-12, produced by R6/2 compared with wild-type cells (Figs. 1A and S1A). Due to the small volumes of fluid available, yields of CD11b+ cells from murine blood are low. To try to identify an alternative source of myeloid cells, other tissues containing monocyte populations were obtained. This also allowed a wider characterisation of HD myeloid cell function. Spleen and bone marrow myeloid cells were isolated and cultured, then primed with IFNγ and stimulated with LPS for 24 h, as before. Spleen myeloid cells from R6/2 mice produced elevated levels of IL-1β and IL-12 compared with wild-type cells, whilst levels of IL-6, TNFα and IL-10 were unchanged (Figs. 1B and S1B). R6/2 bone marrow myeloid cells showed no differences in cytokine production compared with wild-type cells (Figs. 1C and S1C). Therefore, increased cytokine production by hyper-responsive myeloid cells, as seen in HD patients' monocytes and macrophages, was dependent on the tissue origin of myeloid cells in R6/2 mice.


Characterisation of immune cell function in fragment and full-length Huntington's disease mouse models.

Träger U, Andre R, Magnusson-Lind A, Miller JR, Connolly C, Weiss A, Grueninger S, Silajdžić E, Smith DL, Leavitt BR, Bates GP, Björkqvist M, Tabrizi SJ - Neurobiol. Dis. (2014)

Altered cytokine production by myeloid cells from R6/2 mice. (A) CD11b+ myeloid cells were isolated from pooled blood samples obtained from 12-week old R6/2 or wild-type mice. Cell cultures primed with IFNγ and stimulated with LPS for 24 h showed significant differences in IL-6 and TNFα production. (B) R6/2 spleen and (C) R6/2 bone marrow myeloid cells were isolated using anti-CD11b magnetic beads, and individual cultures from each mouse were stimulated with IFNγ and LPS for 24 h. Measuring cytokine production in supernatants by multiplex ELISA, R6/2 spleen myeloid cells demonstrated changes in IL-1β levels, whilst bone marrow myeloid cells from R6/2 mice showed the same cytokine levels as wild-type cells. Graphs show mean concentrations corrected to protein content ± SEM. Unpaired two-tailed Student's t-tests. (A) n = technical replicates of pooled samples from different mice, (B + C) n = biological replicates representing individual mice.
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Related In: Results  -  Collection

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f0005: Altered cytokine production by myeloid cells from R6/2 mice. (A) CD11b+ myeloid cells were isolated from pooled blood samples obtained from 12-week old R6/2 or wild-type mice. Cell cultures primed with IFNγ and stimulated with LPS for 24 h showed significant differences in IL-6 and TNFα production. (B) R6/2 spleen and (C) R6/2 bone marrow myeloid cells were isolated using anti-CD11b magnetic beads, and individual cultures from each mouse were stimulated with IFNγ and LPS for 24 h. Measuring cytokine production in supernatants by multiplex ELISA, R6/2 spleen myeloid cells demonstrated changes in IL-1β levels, whilst bone marrow myeloid cells from R6/2 mice showed the same cytokine levels as wild-type cells. Graphs show mean concentrations corrected to protein content ± SEM. Unpaired two-tailed Student's t-tests. (A) n = technical replicates of pooled samples from different mice, (B + C) n = biological replicates representing individual mice.
Mentions: To determine whether monocytes from HD mouse models are hyper-reactive like those from HD patient blood (Träger et al., 2014), CD11b+ cells from 12-week old R6/2 mice were isolated using magnetic cell sorting and seeded in culture, then primed with IFNγ and stimulated with LPS for 24 h. Analysis of culture supernatants using a multiplex ELISA platform showed an increase in IL-6 and TNFα levels, but not IL-1β, IL-10 and IL-12, produced by R6/2 compared with wild-type cells (Figs. 1A and S1A). Due to the small volumes of fluid available, yields of CD11b+ cells from murine blood are low. To try to identify an alternative source of myeloid cells, other tissues containing monocyte populations were obtained. This also allowed a wider characterisation of HD myeloid cell function. Spleen and bone marrow myeloid cells were isolated and cultured, then primed with IFNγ and stimulated with LPS for 24 h, as before. Spleen myeloid cells from R6/2 mice produced elevated levels of IL-1β and IL-12 compared with wild-type cells, whilst levels of IL-6, TNFα and IL-10 were unchanged (Figs. 1B and S1B). R6/2 bone marrow myeloid cells showed no differences in cytokine production compared with wild-type cells (Figs. 1C and S1C). Therefore, increased cytokine production by hyper-responsive myeloid cells, as seen in HD patients' monocytes and macrophages, was dependent on the tissue origin of myeloid cells in R6/2 mice.

Bottom Line: Inflammation is a growing area of research in neurodegeneration.However, bone marrow CD11b(+) cells did not show the same hyper-responsive phenotype as spleen and blood cells.Taken together, these results show significant promise for these mouse models to be used to study targeting innate immune pathways identified in human cells, thereby helping to understand the role the peripheral immune system plays in HD progression.

View Article: PubMed Central - PubMed

Affiliation: UCL Institute of Neurology, Dept. of Neurodegenerative Disease, London, UK.

No MeSH data available.


Related in: MedlinePlus