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HLA-DRα1-mMOG-35-55 treatment of experimental autoimmune encephalomyelitis reduces CNS inflammation, enhances M2 macrophage frequency, and promotes neuroprotection.

Benedek G, Meza-Romero R, Jordan K, Keenlyside L, Offner H, Vandenbark AA - J Neuroinflammation (2015)

Bottom Line: We here demonstrate that DRα1-mMOG-35-55 could effectively treat EAE in MHC-mismatched C57BL/6 mice by reducing CNS inflammation, potentially mediated in part through an increased frequency of M2 monocytes in the spinal cord.Microarray analysis of spinal cord tissue from DRα1-mMOG-35-55-treated vs. vehicle control mice with EAE revealed decreased expression of a large number of pro-inflammatory genes including CD74, NLRP3, and IL-1β and increased expression of genes involved in myelin repair (MBP) and neuroregeneration (HUWE1).These findings indicate that the DRα1-mMOG-35-55 construct retains therapeutic, anti-inflammatory, and neuroprotective activities during treatment of EAE across MHC disparate barriers.

View Article: PubMed Central - PubMed

Affiliation: Neuroimmunology Research, VA Portland Health Care System, Portland, OR, USA.

ABSTRACT

Background: DRα1-mouse(m)MOG-35-55, a novel construct developed in our laboratory as a simpler and potentially less immunogenic alternative to two-domain class II constructs, was shown previously to target the MIF/CD74 pathway and to reverse clinical and histological signs of experimental autoimmune encephalomyelitis (EAE) in DR*1501-Tg mice in a manner similar to the parent DR2β1-containing construct.

Methods: In order to determine whether DRα1-mMOG-35-55 could treat EAE in major histocompatibility complex (MHC)-mismatched mice and to evaluate the treatment effect on central nervous system (CNS) inflammation, C57BL/6 mice were treated with DRα1-mMOG-35-55. In addition, gene expression profile was analyzed in spinal cords of EAE DR*1501-Tg mice that were treated with DRα1-mMOG-35-55.

Results: We here demonstrate that DRα1-mMOG-35-55 could effectively treat EAE in MHC-mismatched C57BL/6 mice by reducing CNS inflammation, potentially mediated in part through an increased frequency of M2 monocytes in the spinal cord. Microarray analysis of spinal cord tissue from DRα1-mMOG-35-55-treated vs. vehicle control mice with EAE revealed decreased expression of a large number of pro-inflammatory genes including CD74, NLRP3, and IL-1β and increased expression of genes involved in myelin repair (MBP) and neuroregeneration (HUWE1).

Conclusion: These findings indicate that the DRα1-mMOG-35-55 construct retains therapeutic, anti-inflammatory, and neuroprotective activities during treatment of EAE across MHC disparate barriers.

No MeSH data available.


Related in: MedlinePlus

DRα1-mMOG-35-55 treatment reduces the number and activation state of CNS-infiltrating cells. a Absolute lymphocyte numbers in the spinal cord (n = 8) and brain (n = 4) from DRα1-MOG-35-55- or vehicle-treated C57BL/6 male WT mice with EAE. b Absolute numbers of CD11b+CD45high and CD3+ T cells in spinal cords. c CD74 cell surface expression on CD11b+CD45high cells in spinal cords. d Brain lymphocytes from DRα1-mMOG-35-55-treated (n = 4) and vehicle-treated (n = 4) mice were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 4 h. CD4+ T cells were evaluated for the intracellular expression of IFN-γ or IL-17 by flow cytometry. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test
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Fig4: DRα1-mMOG-35-55 treatment reduces the number and activation state of CNS-infiltrating cells. a Absolute lymphocyte numbers in the spinal cord (n = 8) and brain (n = 4) from DRα1-MOG-35-55- or vehicle-treated C57BL/6 male WT mice with EAE. b Absolute numbers of CD11b+CD45high and CD3+ T cells in spinal cords. c CD74 cell surface expression on CD11b+CD45high cells in spinal cords. d Brain lymphocytes from DRα1-mMOG-35-55-treated (n = 4) and vehicle-treated (n = 4) mice were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 4 h. CD4+ T cells were evaluated for the intracellular expression of IFN-γ or IL-17 by flow cytometry. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test

Mentions: We reported previously that two-domain partial MHC class II constructs could reduce EAE severity and CNS inflammation by reducing the number and frequency of activated cells in DR*1501-Tg mice. In order to evaluate if DRα1-mMOG-35-55 treatment of C57BL/6 mice had a similar effect, mononuclear cells were isolated from brains and spinal cords from DRα1-mMOG-35-55- or vehicle-treated mice 5 days post-EAE treatment. The absolute number of mononuclear cells was reduced both in the brain (8.25 × 104 ± 1.2 vs. 18.75 × 104 ± 3.2, p < 0.001) and spinal cord (0.64 × 106 ± 0.4 vs. 1.76 × 106 ± 1.15, p < 0.01), respectively, of DRα1-mMOG-35-55-treated vs. vehicle-treated mice with EAE (Fig. 4a). This reduction was reflected in the absolute number of infiltrating monocytes and activated resident microglia (CD11b+CD45hi) as well as CD3+ T cells in the spinal cord (p < 0.05, Fig. 4b). In addition, the CD74 expression level on CD11b+CD45hi cells was significantly lower in the spinal cords of DRα1-mMOG-35-55- vs. vehicle-treated mice (p < 0.05, Fig. 4c), in accordance with effects of the parent DR*1501 β1α1-mMOG-35-55 construct in DR*1501-Tg mice [30]. Furthermore, there was a significantly lower frequency of PMA/ionomycin-stimulated IL-17+ T cells but not CD4+ IFN-γ+ Τ cells from DRα1-mMOG-35-55-treated mice compared with stimulated cells from vehicle-treated mice (p < 0.05, Fig. 4d).Fig. 4


HLA-DRα1-mMOG-35-55 treatment of experimental autoimmune encephalomyelitis reduces CNS inflammation, enhances M2 macrophage frequency, and promotes neuroprotection.

Benedek G, Meza-Romero R, Jordan K, Keenlyside L, Offner H, Vandenbark AA - J Neuroinflammation (2015)

DRα1-mMOG-35-55 treatment reduces the number and activation state of CNS-infiltrating cells. a Absolute lymphocyte numbers in the spinal cord (n = 8) and brain (n = 4) from DRα1-MOG-35-55- or vehicle-treated C57BL/6 male WT mice with EAE. b Absolute numbers of CD11b+CD45high and CD3+ T cells in spinal cords. c CD74 cell surface expression on CD11b+CD45high cells in spinal cords. d Brain lymphocytes from DRα1-mMOG-35-55-treated (n = 4) and vehicle-treated (n = 4) mice were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 4 h. CD4+ T cells were evaluated for the intracellular expression of IFN-γ or IL-17 by flow cytometry. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test
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Fig4: DRα1-mMOG-35-55 treatment reduces the number and activation state of CNS-infiltrating cells. a Absolute lymphocyte numbers in the spinal cord (n = 8) and brain (n = 4) from DRα1-MOG-35-55- or vehicle-treated C57BL/6 male WT mice with EAE. b Absolute numbers of CD11b+CD45high and CD3+ T cells in spinal cords. c CD74 cell surface expression on CD11b+CD45high cells in spinal cords. d Brain lymphocytes from DRα1-mMOG-35-55-treated (n = 4) and vehicle-treated (n = 4) mice were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 4 h. CD4+ T cells were evaluated for the intracellular expression of IFN-γ or IL-17 by flow cytometry. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test
Mentions: We reported previously that two-domain partial MHC class II constructs could reduce EAE severity and CNS inflammation by reducing the number and frequency of activated cells in DR*1501-Tg mice. In order to evaluate if DRα1-mMOG-35-55 treatment of C57BL/6 mice had a similar effect, mononuclear cells were isolated from brains and spinal cords from DRα1-mMOG-35-55- or vehicle-treated mice 5 days post-EAE treatment. The absolute number of mononuclear cells was reduced both in the brain (8.25 × 104 ± 1.2 vs. 18.75 × 104 ± 3.2, p < 0.001) and spinal cord (0.64 × 106 ± 0.4 vs. 1.76 × 106 ± 1.15, p < 0.01), respectively, of DRα1-mMOG-35-55-treated vs. vehicle-treated mice with EAE (Fig. 4a). This reduction was reflected in the absolute number of infiltrating monocytes and activated resident microglia (CD11b+CD45hi) as well as CD3+ T cells in the spinal cord (p < 0.05, Fig. 4b). In addition, the CD74 expression level on CD11b+CD45hi cells was significantly lower in the spinal cords of DRα1-mMOG-35-55- vs. vehicle-treated mice (p < 0.05, Fig. 4c), in accordance with effects of the parent DR*1501 β1α1-mMOG-35-55 construct in DR*1501-Tg mice [30]. Furthermore, there was a significantly lower frequency of PMA/ionomycin-stimulated IL-17+ T cells but not CD4+ IFN-γ+ Τ cells from DRα1-mMOG-35-55-treated mice compared with stimulated cells from vehicle-treated mice (p < 0.05, Fig. 4d).Fig. 4

Bottom Line: We here demonstrate that DRα1-mMOG-35-55 could effectively treat EAE in MHC-mismatched C57BL/6 mice by reducing CNS inflammation, potentially mediated in part through an increased frequency of M2 monocytes in the spinal cord.Microarray analysis of spinal cord tissue from DRα1-mMOG-35-55-treated vs. vehicle control mice with EAE revealed decreased expression of a large number of pro-inflammatory genes including CD74, NLRP3, and IL-1β and increased expression of genes involved in myelin repair (MBP) and neuroregeneration (HUWE1).These findings indicate that the DRα1-mMOG-35-55 construct retains therapeutic, anti-inflammatory, and neuroprotective activities during treatment of EAE across MHC disparate barriers.

View Article: PubMed Central - PubMed

Affiliation: Neuroimmunology Research, VA Portland Health Care System, Portland, OR, USA.

ABSTRACT

Background: DRα1-mouse(m)MOG-35-55, a novel construct developed in our laboratory as a simpler and potentially less immunogenic alternative to two-domain class II constructs, was shown previously to target the MIF/CD74 pathway and to reverse clinical and histological signs of experimental autoimmune encephalomyelitis (EAE) in DR*1501-Tg mice in a manner similar to the parent DR2β1-containing construct.

Methods: In order to determine whether DRα1-mMOG-35-55 could treat EAE in major histocompatibility complex (MHC)-mismatched mice and to evaluate the treatment effect on central nervous system (CNS) inflammation, C57BL/6 mice were treated with DRα1-mMOG-35-55. In addition, gene expression profile was analyzed in spinal cords of EAE DR*1501-Tg mice that were treated with DRα1-mMOG-35-55.

Results: We here demonstrate that DRα1-mMOG-35-55 could effectively treat EAE in MHC-mismatched C57BL/6 mice by reducing CNS inflammation, potentially mediated in part through an increased frequency of M2 monocytes in the spinal cord. Microarray analysis of spinal cord tissue from DRα1-mMOG-35-55-treated vs. vehicle control mice with EAE revealed decreased expression of a large number of pro-inflammatory genes including CD74, NLRP3, and IL-1β and increased expression of genes involved in myelin repair (MBP) and neuroregeneration (HUWE1).

Conclusion: These findings indicate that the DRα1-mMOG-35-55 construct retains therapeutic, anti-inflammatory, and neuroprotective activities during treatment of EAE across MHC disparate barriers.

No MeSH data available.


Related in: MedlinePlus