top
Please input keywords
EAE model

Introduction of experimental autoimmune encephalomyelitis (EAE) model


Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system that leads to encephalitis and demyelination. MS is considered an autoimmune disease caused by autoreactive T cells with symptoms including muscle rigidity and paralysis, visual disturbance and blindness, sensory loss and ataxia, and is characterized by recurrent relapses. At present, there are many different animal models of MS, of which the experimental autoimmune encephalomyelitis (EAE) model is widely used in the study of multiple sclerosis due to its pathological characteristics of inflammation and demyelination similar to MS.
In rodents such as mice and rats, EAE models can be induced by immunization with spinal cord homogenates, purified myelin, myelin proteins (e.g., myelin basic protein MBP, protein lipoprotein PLP, and myelin oligodendrocyte glycoprotein MOG), or peptides of these proteins. This may be because myelin-specific T cells are activated in the periphery, cross the blood-brain barrier into the central nervous system and are reactivated, triggering a series of inflammatory responses, leading to demyelination and axonal cell apoptosis, ultimately leading to nerve injury and loss of function. Clinical symptoms are assessed using a standardized scoring system, which measures the degree of induction of the disease, with focal demyelination and inflammatory leukocyte infiltration seen in the stained sections of the pathological tissue.
The MOG-induced EAE disease model protocol described here was established in C57BL/6 mice and in B-hIL17A humanized mice developed by Biocytogen, andcan be used for pharmacodynamic evaluation of MS-related drugs.


Establishment of EAE mouse model


EAE Induction in C57BL/6 Mice

Model Building Schematic


from clipboard


Clinical Score


from clipboard

MOG (define) immunization was used in 10-week-old C57BL/6 female mice (n = 5), which were given PTX (define) on the day of immunization and the following day, respectively. (A) Body weight change of animals in each group. (B) Clinical scores of animals in each group. ompared with the untreatedgroup (G1-Vehicle), the MOG-treatedgroup (G2) had tail weakness, lameness, hind limb paralysis and other symptoms, resulting in an increased  clinical score. This demonstrates that EAE was successfully induced in C57BL/6 mice. Data are shown as mean ± SEM. MOG: myelin oligodendrocyte glycoprotein. PTX: Pertussis toxin.


Pathological analysis


from clipboard


Local inflammatory responses in the central nervous system (CNS) of EAE model mice. Spinal cords were taken on day 35 after MOG and PTX immunization, and tissue sections were stained with H&E (A, B) and detected by immunofluorescence (IF) (C, D) (green: MBP; blue: DAPI). nflammatory cell infiltration (DAPI+ cells) was significantly increased and myelin protein was significantly reduced in the model group. This suggests that the EAE disease model was successfully induced in C57BL/6 mice.


Induction of EAE disease model in B-hIL17A mice

Experimental Schematic


from clipboard


Clinical Score


from clipboard


10-week-old B-hIL17A female mice (n = 5) were immunized with MOG, and given PTX on the day of immunization and the following day, respectively. (A) ody weight of animals in each group. (B) Clinical scores of animals in each group. ompared with the non-model group (G1-Vehicle), the model group (G2-MOG) exhibitedtail weakness, lameness, hind limb paralysis and other symptoms, resulting in an increase in  clinical score. This shows that the EAE disease model was successfully induced in B-hIL17A mice. Data are shown as mean ± SEM.


Pathological analysis


from clipboard


Local inflammatory responses in the central nervous system (CNS) ofEAE-induced B-hIL17Amice. Spinal cords were isolated on day 45 after MOG and PTX immunization, and tissue sections were stained with H&E (A, B) and detected by immunofluorescence (IF) (C, D) (green: MBP; blue: DAPI). The results showed that inflammatory cell infiltration was significantly increased and myelin protein was significantly reduced in the model group (G2-MOG). This indicates that the EAE disease model was successfully induced in B-hIL17A mice.


Lymph node cells by flow cytometry (FACS)


from clipboard


Lymph node cells from MOG and PTX-immunized B-hIL17A female mice (n = 5) were used to detect the number of IL17A+ and IFN-γ+ cells by flow cytometry after treatment with brefeldin A (BFA) and simultaneous stimulation with myristoyl phorbol ethyl ester (PMA) and ionomycin (Ionomycin) for 6 hours. (A) Assessment of cytokine expression  of lymph node cells detected by FACS. (B) Numbers of IL17A + CD3 + CD4 + T cells and IFN-γ + T cells in each group of animals. The results indicate that the number of IFN-γ + T cells in lymph nodes was increased and the number of IL17A + CD3 + CD4 + T cells was significantly increased in B-hIL17A mice after immunization with MOG/CFA.


Detection of central nervous system (CNS) brain cells by flow cytometry (FACS)


from clipboard

CNS brain cells from MOG and PTX-immunized B-hIL17A female mice (n = 5) were treated with brefeldin A (BFA) and stimulated simultaneously with myristoyl phorbol ethyl ester (PMA) and ionomycin (Ionomycin) for 6 h. The number of IL17A + and IFN-γ + cells was measured by flow cytometry. (Fig. A) Assessment of cytokine expressing lymph node cells detected by FACS. (B) Numbers of IL17A+ CD3+ CD4+ T cells and IFN-γ+ T cells in each group of animals. The results showed that IL17A+ CD3+ CD4+ T cells and IFN-γ+ T cells in brain cells were significantly increased in B-hIL17A mice after immunization with MOG and PTX.


product list:


Product name

Product number

B-hIL17A mice

110053

B-hTNFA mice

110002

References


1. Gaffen, S.L., Jain, R., Garg, A.V. & Cua, D.J. The IL-23-IL-17 immune axis: from mechanisms to therapeutic testing. Nat Rev Immunol 14, 585-600 (2014).

2. Iwakura, Y. & Ishigame, H. The IL-23/IL-17 axis in inflammation. J Clin Invest 116, 1218-1222 (2006).

3. Kuwabara, T., Ishikawa, F., Kondo, M. & Kakiuchi, T. The Role of IL-17 and Related Cytokines in Inflammatory Autoimmune Diseases. Mediators Inflamm 2017, 3908061 (2017).