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Regulatory lymphocytes are key factors in MHC-independent resistance to EAE.

Marín N, Mecha M, Espejo C, Mestre L, Eixarch H, Montalban X, Álvarez-Cermeño JC, Guaza C, Villar LM - J Immunol Res (2014)

Bottom Line: We aimed to explore MHC-independent mechanisms inducing resistance to EAE.We studied T cell proliferation, regulatory and effector cell subpopulations, intracellular and serum cytokine patterns, and titers of anti-MOG serum antibodies.Expansion of regulatory B and T cells contributes to the induction of resistance to EAE by an MHC-independent mechanism.

View Article: PubMed Central - PubMed

Affiliation: Multiple Sclerosis Unit, Immunology and Neurology Departments, Hospital Universitario Ramón y Cajal, IRYCIS, Carretera de Colmenar Km 9.100, 28034 Madrid, Spain ; Red Española de Esclerosis Múltiple (REEM), RETICS, Fondo de Investigación Sanitaria, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain.

ABSTRACT

Background and objectives: Resistant and susceptible mouse strains to experimental autoimmune encephalomyelitis (EAE), an inducible demyelinating experimental disease serving as animal model for multiple sclerosis, have been described. We aimed to explore MHC-independent mechanisms inducing resistance to EAE.

Methods: For EAE induction, female C57BL/6 (susceptible strain) and CD1 (resistant outbred strain showing heterogeneous MHC antigens) mice were immunized with the 35-55 peptide of myelin oligodendrocyte glycoprotein (MOG35-55). We studied T cell proliferation, regulatory and effector cell subpopulations, intracellular and serum cytokine patterns, and titers of anti-MOG serum antibodies.

Results: Upon immunization with MOG35-55, T lymphocytes from susceptible mice but not that of resistant strain were capable of proliferating when stimulated with MOG35-55. Accordingly, resistant mice experienced a rise in regulatory B cells (P=0.001) and, to a lower extent, in regulatory T cells (P=0.02) compared with C57BL/6 susceptible mice. As a consequence, MOG35-55-immunized C57BL/6 mice showed higher percentages of CD4+ T cells producing both IFN-gamma (P=0.02) and IL-17 (P=0.009) and higher serum levels of IL-17 (P=0.04) than resistant mice.

Conclusions: Expansion of regulatory B and T cells contributes to the induction of resistance to EAE by an MHC-independent mechanism.

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(a) Intracellular production of interleukin-17 (IL-17) by spleen CD4+ T cells. We analyzed these cells in series of seven mice per condition. Splenocytes from C57BL/6 and CD1 mice were cultured in presence of phorbol-12-myristate-13-acetate (PMA) and ionomycin during four hours and the percentage of CD4+ T cells showing intracellular production of IL-17 was analyzed by flow cytometry. We studied nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG) and with PLP139–151 (PLP). Only C57BL/6 mice that developed EAE after immunization with MOG35–55 displayed a significant increase in the percentage of CD4+ T cells producing IL-17. (b) Representative dot plots of IL-17 intracellular production by CD4+ T cells from MOG35–55-immunized CD1 and C57BL/6 mice are shown. (c) Serum concentrations of IL-17A in nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG).
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fig5: (a) Intracellular production of interleukin-17 (IL-17) by spleen CD4+ T cells. We analyzed these cells in series of seven mice per condition. Splenocytes from C57BL/6 and CD1 mice were cultured in presence of phorbol-12-myristate-13-acetate (PMA) and ionomycin during four hours and the percentage of CD4+ T cells showing intracellular production of IL-17 was analyzed by flow cytometry. We studied nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG) and with PLP139–151 (PLP). Only C57BL/6 mice that developed EAE after immunization with MOG35–55 displayed a significant increase in the percentage of CD4+ T cells producing IL-17. (b) Representative dot plots of IL-17 intracellular production by CD4+ T cells from MOG35–55-immunized CD1 and C57BL/6 mice are shown. (c) Serum concentrations of IL-17A in nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG).

Mentions: We did not find differences in the levels of IgM or IgG anti-MOG35–55 antibodies or in total CD4+ and CD8+ T cell percentages between the resistant and the susceptible mice (Table 1). To further discriminate different T cell responses, we analyzed Th1 and Th17 subsets. We explored the percentage of CD4+ T cells showing intracellular production of IFN-gamma (Figure 4) and IL-17 (Figure 5) upon stimulation with phorbol-12-myristate-13-acetate (PMA) and ionomycin. Only susceptible mice that developed EAE in response to MOG35–55 showed significant increases in the secretion of IFN-gamma (P = 0.02) and IL-17 (P = 0.009) compared to resistant mice. By contrast, resistant mice were unable to mount any of these responses after the same immunization protocol. In addition, when studying serum inflammatory cytokines, we detected a significant increase of IL-17 in susceptible mice immunized with MOG35–55 (Figure 4(c)). This shows that downmodulation of regulatory B and T cells is related to the induction of Th1/Th17 responses in MOG-induced EAE.


Regulatory lymphocytes are key factors in MHC-independent resistance to EAE.

Marín N, Mecha M, Espejo C, Mestre L, Eixarch H, Montalban X, Álvarez-Cermeño JC, Guaza C, Villar LM - J Immunol Res (2014)

(a) Intracellular production of interleukin-17 (IL-17) by spleen CD4+ T cells. We analyzed these cells in series of seven mice per condition. Splenocytes from C57BL/6 and CD1 mice were cultured in presence of phorbol-12-myristate-13-acetate (PMA) and ionomycin during four hours and the percentage of CD4+ T cells showing intracellular production of IL-17 was analyzed by flow cytometry. We studied nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG) and with PLP139–151 (PLP). Only C57BL/6 mice that developed EAE after immunization with MOG35–55 displayed a significant increase in the percentage of CD4+ T cells producing IL-17. (b) Representative dot plots of IL-17 intracellular production by CD4+ T cells from MOG35–55-immunized CD1 and C57BL/6 mice are shown. (c) Serum concentrations of IL-17A in nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4020375&req=5

fig5: (a) Intracellular production of interleukin-17 (IL-17) by spleen CD4+ T cells. We analyzed these cells in series of seven mice per condition. Splenocytes from C57BL/6 and CD1 mice were cultured in presence of phorbol-12-myristate-13-acetate (PMA) and ionomycin during four hours and the percentage of CD4+ T cells showing intracellular production of IL-17 was analyzed by flow cytometry. We studied nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG) and with PLP139–151 (PLP). Only C57BL/6 mice that developed EAE after immunization with MOG35–55 displayed a significant increase in the percentage of CD4+ T cells producing IL-17. (b) Representative dot plots of IL-17 intracellular production by CD4+ T cells from MOG35–55-immunized CD1 and C57BL/6 mice are shown. (c) Serum concentrations of IL-17A in nonimmunized mice (NI) and mice immunized with MOG35–55 (MOG).
Mentions: We did not find differences in the levels of IgM or IgG anti-MOG35–55 antibodies or in total CD4+ and CD8+ T cell percentages between the resistant and the susceptible mice (Table 1). To further discriminate different T cell responses, we analyzed Th1 and Th17 subsets. We explored the percentage of CD4+ T cells showing intracellular production of IFN-gamma (Figure 4) and IL-17 (Figure 5) upon stimulation with phorbol-12-myristate-13-acetate (PMA) and ionomycin. Only susceptible mice that developed EAE in response to MOG35–55 showed significant increases in the secretion of IFN-gamma (P = 0.02) and IL-17 (P = 0.009) compared to resistant mice. By contrast, resistant mice were unable to mount any of these responses after the same immunization protocol. In addition, when studying serum inflammatory cytokines, we detected a significant increase of IL-17 in susceptible mice immunized with MOG35–55 (Figure 4(c)). This shows that downmodulation of regulatory B and T cells is related to the induction of Th1/Th17 responses in MOG-induced EAE.

Bottom Line: We aimed to explore MHC-independent mechanisms inducing resistance to EAE.We studied T cell proliferation, regulatory and effector cell subpopulations, intracellular and serum cytokine patterns, and titers of anti-MOG serum antibodies.Expansion of regulatory B and T cells contributes to the induction of resistance to EAE by an MHC-independent mechanism.

View Article: PubMed Central - PubMed

Affiliation: Multiple Sclerosis Unit, Immunology and Neurology Departments, Hospital Universitario Ramón y Cajal, IRYCIS, Carretera de Colmenar Km 9.100, 28034 Madrid, Spain ; Red Española de Esclerosis Múltiple (REEM), RETICS, Fondo de Investigación Sanitaria, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain.

ABSTRACT

Background and objectives: Resistant and susceptible mouse strains to experimental autoimmune encephalomyelitis (EAE), an inducible demyelinating experimental disease serving as animal model for multiple sclerosis, have been described. We aimed to explore MHC-independent mechanisms inducing resistance to EAE.

Methods: For EAE induction, female C57BL/6 (susceptible strain) and CD1 (resistant outbred strain showing heterogeneous MHC antigens) mice were immunized with the 35-55 peptide of myelin oligodendrocyte glycoprotein (MOG35-55). We studied T cell proliferation, regulatory and effector cell subpopulations, intracellular and serum cytokine patterns, and titers of anti-MOG serum antibodies.

Results: Upon immunization with MOG35-55, T lymphocytes from susceptible mice but not that of resistant strain were capable of proliferating when stimulated with MOG35-55. Accordingly, resistant mice experienced a rise in regulatory B cells (P=0.001) and, to a lower extent, in regulatory T cells (P=0.02) compared with C57BL/6 susceptible mice. As a consequence, MOG35-55-immunized C57BL/6 mice showed higher percentages of CD4+ T cells producing both IFN-gamma (P=0.02) and IL-17 (P=0.009) and higher serum levels of IL-17 (P=0.04) than resistant mice.

Conclusions: Expansion of regulatory B and T cells contributes to the induction of resistance to EAE by an MHC-independent mechanism.

Show MeSH
Related in: MedlinePlus