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The Clinical and Basic Science Study of B7H1 Immunoregulation in Multiple Sclerosis

Author: LiYuJing
Tutor: DongWanLi
School: Suzhou University
Course: Neurology
Keywords: B7H1 multiple sclerosis EAE
CLC: R744.51
Type: Master's thesis
Year: 2011
Downloads: 29
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Abstract


ObjectiveTo investigate the expression of B7H1 on the mononuclear cells in peripheral blood of the patients with multiple sclerosis using flow cytometry; To investigate the expression of B7H1 on the central nervous system and spleen cells of the animal model EAE for multiple sclerosis with C3H10 cells transplated using immunofluorescence and flow cytometry;To explor B7H1 immunoregulation in multiple sclerosis.MethodsThe study was designed in the first hospital affiliated to Soochow University between September 2009 and February 2011. Twenty-five patients with multiple sclerosis were divided into two groups, acute and stable.Cases of acute patients were given hormones, azathioprine or other immunosuppressive therapy for one month.20 subjects matched by age and gender who made health examination in the hospital served as controls. After overnightfasting, a single blood sample was obtained. CD4+T cells, CD14~+ mononuclear macrophages and the expression of B7H1 on these cells were measured using flow cytometry.Tirty C57BL/6J mice were devided into three groups randomly: the EAE group, the cells transplated group and the normal controls. The mice in EAE group and transplated group were immunized subcutaneously with MOG35-55 emulsified in CFA that was further enriched with Mycobacterium tuberculosis. The normal mice were injected i.v. with PBS. Expression of B7H1 on C3H10T1 / 2 (C3H10) was detected using flow cytometry. C3H10 stained with GSFE were transferred i.v. into the transplated group and analyzed by fluorescence microscopy for its localization. The clinical course and body weight of the three groups were evaluated daily. Spinal cords were removed; the slices were made and then treated with HE stain and luxol fast blue stain for light microscopy. The expression of B7H1, astrocytes and microglial cells were investigated using immunofluorescence. The expression of CD4+B7H1~+T cells and CD14~+B7H1~+ mononuclear macrophages in spleen were investigated using flow cytometry.Results1. CD4+B7H1~+T cells and CD14~+B7H1~+ mononuclear macrophages were significantly higher in the peripheral blood of patients with multiple sclerosis compared to the normal controls,and the B7H1~+ immune cells in SMS patients were higher than in AMS patients;2. CD4+B7H1~+T cells and CD14~+B7H1~+ mononuclear macrophages in the peripheral blood of patients can be reduced after immunosuppressive therapy;3. The EAE mice immunized had gradual emergence of weight loss,motor retardation and other symptoms of EAE, obvious inflammatory cells infiltration and demyelination can be seen in spinal cord of the EAE mice, the transplanted mice and the normal controls had no such performance;4. B7H1, astrocytes, microglial cells in spinal cord and CD4+ B7H1~+ T cells ,CD14~+ B7H1~+ mononuclear macrophages in peripheral spleen of EAE mice were significantly higher compared to the transplated mice and the normal controls;5. C3H10 had B7H1 expression on surface; they could migrate and gather in the spinal cord lesion sites of the transplated mice.Conclusions1. B7H1 plays a negative immunomodulation role in the peripheral blood of the patients with multiple sclerosis by CD4+T cells and CD14~+ mononuclear macrophages.2. The immunosuppressive therapy could improve the symptoms of multiple sclerosis by the negative immunomodulation of B7H1.3. B7H1 plays a negative immunomodulation role in central nervous system of the EAE mice by astrocytes and microglial cells; B7H1 plays a negative immunomodulation role in spleen of the EAE mice by CD4+T cells and CD14~+ mononuclear macrophages.4. C3H10 can inhibit EAE, possibly by the negative immunomodulation of B7H1. 5. B7H1 can be a target of occurring, mitigation, severity and therapeutics of multiple sclerosis and the target molecule of its treatment; embryonic mesenchymal stem cell transplantation may also be used as a new way in the treatment of multiple sclerosis.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Spinal cord disease > Demyelinating disease > Multiple sclerosis
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