Effect of doxycycline on Th1/Th2 cell balance and related cytokines in experimental allergic encephalomyelitis rats / 重庆医学

Objective To investigate the effect of doxycycline on the Th1/Th2 cell balance in experimental allergic encepha-lomyelitis(EAE)rats.Methods Forty female Wistar rats were randomly divided into the EAE control group,low,medium and high does DOX treatment groups,10 cases in each group.The onset situation in rats was observed.The IL-4 and IFN-γlevels secreted by peripheral blood mononuclear cells (PBMC)at the peak stage were detected.The levels of IL-1β,IL-10,TNF-αin brain tissue,and the albumin content in cerebrospinal fluid and serum were detected.The QA value was calculated.Results In each DOX group,the clinical symptoms of rats were alleviated compared with the EAE control group.In each DOX group,the PBMC secreting IFN-γlev-el and IFN-γ/IL-4 ratio in the onset peak stage were lower than those in the EAE control group,while the IL-4 level was higher than that in the EAE control group(P 0.05),but the difference between other DOX groups had statistical significance(P <0.01).The IL-1βand TNF-αlevels of brain tissue and QA value during onset peak stage in various doses DOX groups were decreased compared with the EAE control group,while the IL-10 level was increased compared with the EAE control group(P <0.05).With the DOX dose increasing,the levels of IL-1β,TNF-αand QA value in various doses DOX groups became lower,the IL-10 level became high-er,there was statistically significant difference among various doses DOX groups (P <0.05 ).Conclusion DOX can obviously alle-viate the clinical symptoms of EAE rats,its mechanism may be related with that DOX could decrease the level of Th1 cytokine and increase the level of Th2 cytokine,correct the Th1/Th2 cell balance,thus protect the blood brain barrier(BBB).

Establish and optimize the model of experimental allergic encephalomyelitis in mice / 中华行为医学与脑科学杂志

Objective To explore the appropriate dosage of drugs inducing experimental allergic en cephalomyelitis (EAE) in mice,and evaluate the modified model mice.Methods Different doses of myelin oligodendrocyte glycoprotein (MOG35-55:200 μg,100 μg,50 μg,25 μg),together with different doses of inactivation of mycobacterium tuberculosis (H37RA:800 μg,250 μg,100 μg) and pertussis toxin (500 ng,200 ng),were used to induce the EAE model.After immunized,the clinical disease severity of EAE mice was measured by the standard EAE grading clinical score daily.The open field test was used to detect the locomotion of mice.The Western blot and immunofluorescence were used to detect the level of myelin basic proteins (MBP) in different brain regions of mice.Results Compared with the EAE mice induced with high-dose drugs,the mice with low-dose drugs (25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) had low neu rological scores.And they displayed normal locomotion compared with the control mice (day 16:group EAE (8.885±0.772) cm/s vs control group (8.933±0.567) cm/s,P＞0.05;day 31:group EAE (11.130±0.630) cm/s vs control group (10.670±0.959) cm/s,P＞0.05;day 55:group EAE (7.686±0.428) cm/s vs control group (8.313±0.918) cm/s,P＞0.05).Moreover,there was a significant decrease of MBP in the parahippocampal cortex (PHC) and fimbria-fornix of EAE mice induced with low-dose of drugs (PHC:group EAE (0.369±0.096) vs control group (1.000±0.163),P＜0.05;fimbria-fomix:group EAE (0.494±0.071) vs control group (1.000±0.143),P＜0.05).Conclusion The EAE mice induced with low-dose drugs(25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) have low neurological scores,normal locomotion,and myelin impairment in the central neuronal system.And it can be used in the cognitive behavioral research of demyelination disease,such as multiple sclerosis.

Effect of leflunomide on peripheral blood CD28/CTLA-4 costimulatory molecules expression in guinea pig with EAE / 中国生化药物杂志

Objective To investigate immunological mechanism underlying leflunomide ( LEF ) against experimental allergic encephalomyelitis ( EAE) by studying the effects of LEF on peripheral blood CD28/CTLA-4 costimulatory molecules expression in guinea pig with EAE.Methods 50 adult female guinea pigs were randomly divided into normal control group,low dose group,EAE control group,middle dose group,and high dose group, 10 guinea pigs in each group respectively.Low, medium and high dose group were treated with LEF 10 mg/kg· D, 20 mg/kg· D and 40 mg/kg· D orally, 1 times/day, to the termination of the experiment.The expression of CD28, CTLA-4 were detected by flow cytometry, incidence was recorded at the same time.Results Compared with EAE control group,high dose and middle dose group incubation period were prolonged, progress period were shorten and neurological dysfunction score decreased(P<0.05).Compared with normal control group, the expression of CD28 of EAE control group increased and the decreased expression of CTLA-4 (P<0.05).High, middle dose treatment group compared with EAE control group CD28 down regulated expression (P<0.05), but the expression of CTLA-4 increased(P<0.05).Among the treatment groups, of CD28 molecules with high dose group decreased significantly(P<0.05), the expression of CTLA-4 molecules with high dose treatment groups upregulated significantly (P<0.05).Correlation analysis showed that the EAE control group, the treatment group the CD28/CTLA-4 ratio in peripheral blood and nerve dysfunction score is proportional ( r=0.85, P=0.002; r=0.77, P=0.000).Conclusion LEF can reduce EAE guinea pig neurological symptoms, the better and the higher dose effect.Its mechanism may be through down-regulation of CD28 molecules in peripheral blood, upregulation of the expression of CTLA-4 molecule.so as to reduce the inflammatory response, relieve the clinical symptoms.

Effect of ulinastatin on the nerve regeneration of mice encephalomyelitis / 中国生化药物杂志

Objective To investigate the effects of ulinastatin(UTI)on the nerve regeneration of mice experimental allergic encephalomyelitis and the expression of related factors and protein.Methods Forty mice were randomly divided into four groups:ulinastatin group(U),atorvastatin group(A), empty control group(C)and normal control group(N).The experimental autoimmune encephalomyelitis(EAE)in mice was constructed by Freund's complete adjuvant and MOG35-55 polypeptide.Histopathological changes were observed by HE,LFB and Bielschowsky stained at the 3rd week and 4th week after immunized of each group.The expressions of CD4 +T cells were estimated by immunohistochemical method.The expression of myelin basic protein (MBP),brain-derived neurotrophic factor(BDNF),growth associated protein-43(GAP-43),2',3'-cyclic nucleotide-3'-phosphodiesterase(CNP)were detected by Western-blot.Results The largest neurological score of group U was lower than group C,and the difference was statistically significant(P<0.05 ).Pathological features showed that the inflammatory cells,demyelination of spinal cord and axonal injury of group U were lighter than group C.With the duration of treatment,nerve injury decreased.After UTI treatment,the expression of MBP,BDNF,GAP-43,CNP increased.They were statistically significant difference when compared with group C(P<0.05).There was no significant difference between ulinastatin and atorvastatin in the treatment of EAE.Conclusion Ulinastatin could reduce the extent of nerve damage effectively and promote its regeneration which provide a theoretical basis for the clinical treatment of MS.

Expression of myelin basic protein in rat optic nerve with experimental allergic encephalomyelitis / 中华实验眼科杂志

Background Optic neuritis is closely associated with multiple sclerosis (MS).Its pathogenesis is uncompletely clear,and less basic researches are carried out at home and abroad. Objective This study was to reveal the expression of myelin basic protein (MBP) in the optic nerve of rat with experimental allergic encephalomyelitis (EAE) and to provide a theoretical evidence for the research of the relationship of optic neuritis with MS. Methods Fifty clean Wistar rats were randomized into the control group and immune 8,12,18 and 25 days groups.Myelencephalon was collected from 5 guinea pigs to prepare the homogenate and mixed with the isovolumetric complete Freud' s adjuvant (CFA).The 0.5 ml mixed antigen emulsifier was subcutancously injected into the 4 maps together with Bordetella pertussis 0.2 ml under the cutancous of dorsalis pedis at 0 and 48 hours to induce the EAE.Behavior of the rats was evaluated to score the neurological function.The optical nerve sections were prepared 8,12,18 and 25 days after immunology for the histopathological examination,and immunochemistry and Western blot were used to detect the expression of MBP in optic nerve.The use of the animals complied with the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission. Results The disorder of motor nerve was seen 12 days following the immune,and the clinical neural functional scores were significantly higher 12 day and peaked on 18 days myelination and then gradually reduced.The histopathological examination showed that the irregular alignment of neural fiber was found at 12 days,and changes of cellular structure,edema of neural shaft bunch were observed at 18 days.However,the abnormal cells were significantly less 25 days following the immune.Immunochemistry showed that the MBP was expressed mainly in the myelination of optic nerve fibers.The numbers of positive cells for MBP were (115.75±26.49)cells/5 fields at the 12th day,showing a significant lowing in comparison with (167.44±22.49)cells/5 fields of control group (t=4.537,P＜0.05 ).The positive cells were lest at the 18th day with the values ( 75.57 ± 34.54) cells/5 fields ( t =6.362,P＜0.01 ).At the 25th day,positive cells increased to ( 117.63 ± 13.78) cells/5 fields,which still was lower than those of the control group ( t =4.068,P＜0.05 ).Western blot assay illuminated that with the prolong of immune time,MBP/β-actin ratio in optic nerve was gradually reduced and followed the same pattern at the 12th day( t =4.639,P＜0.05 ).At 18 days after immune in comparison with the control group,the expression of MBP/β-actin ratio in optic nerve was the least (t=8.427,P＜0.01). Conclusions MBP can be degraded in rat optic nerve.This is a further evidence that optic neuritis is a severe demyelination disease.It is clearly related to MS.

Erythropoietin ameliorate experimental allergic encephalomyelitis / 中国免疫学杂志

Objective:To investigate the beneficial effect of erythropoietin(EPO)on experimental allergic encephalomyelitis(EAE).Methods: The severity of EAE mice with or without EPO therapy were evaluated through clinical symptoms score and histological observation.Mononuclear cells from spleens,lymph nodes, brains and lumber spinal cords were applied to flow cytometry to detect ratios of CD4~+T subgroup, scales of apoptotic and dead cells, levels of inflammatory cytokines.LDH release of peripheral mononuclear cells were examined after incubating with EPO at different concentrations.Results: EPO ameliorated clinical symptoms and infiltration of pathological inflammatory cells in EAE mice(P＜0.05); Ratio of CD4~+T subgroup, levels of IL-17 and IFN-γ in EAE mice with EPO therapy were significantly lower than those of EAE control group(P＜0.05).No statistical significant differences existed on scales of dead cells, levels of IL-10 and CD4~+ CD25~+ T cells (P＞0.05).EPO at the concentrstions of 1-1 000 U/ml had no effect on LDH release of peripheral mononuclear cells(P＞0.05).Conclusion: EPO play the role of neuroprotecion in EAE mice by the way of decreasing infiltrating inflammatory cells and lowering the levels of IL-17 and IFN-γ.

Establishment of experimental allergic encephalomyelitis in C57BL/6 mice and testing of expressions of Foxp3 and quantifies of CD4~+ CD25~+ regulatory T cells / 中华微生物学和免疫学杂志

Objective To induce experimental allergic encephalomyelitis (EAE) in female C57BL/6 mice with the extracellular domain of myelin oligedendroglia glycoprotein(MOG~(Igd)). Percentages of CD4~+ CD25~+ T cell (Tr) were tested , and also normalized expressions of Foxp3. Methods Molecular cloning technology was used to produce MOG~(Igd) fusion protein. The MOG~(Igd)-TrxA fusion protein and TrxA protein were purified by metal chelate affinity chromatography (MCAC). Mice were injected s. c. in the flank with 300 μg MOG~(Igd) in complete Frcund's adjuvant (CFA) supplemented with 4 μg/μl Mycobacterium tuberculosis. H37Rv. Mice received 0.4 ml emulsion of spinal cord homogenate of guinea pigs (GPSCH) in positive control group, and the same volume emulsiom of TrxA in negative control group, while mice served as normal control received only saline/adjuvant. Mice were monitored two times a day for continuously 30 days by double bind. Clinical scores and histopathology were evaluated. Then, mice were sacrificed. The spinal cord and brain were removed and fixed in buffered formalin. Horizontal sections taken from the central nervous system(CNS) were stained with haematoxylin and eosin (HE), and Kluver-Barrera staining. Also, immunohistochemistry was performed. Percentages of CD4~+ CD25~+ T cells were tested through flow cytometric analysis, and real-time PCR was performed to test normalized expressions of Foxp3 mRNA. Then, correlations between the two were performanced. Results Mice in both MOG group and GPSCH group shew chronic non-remitting course. The onset of disease, time when the most severe clinical symptoms happened and the clinical score between the two groups shew no significant differnces (P>0.05). However, neither in TrxA treated group nor in normal control group did animals exhibit clinical signs of EAE. Histologic sections of the brain and spinal cord taken from affected animals shew perivascular infiltration of mononuclear cells, gliosis, and multifocal demyelination. Lesions scattered throughout the CNS including brainstem, spinal cord, cerebellum, and penventricular white matter. There were significant differences between MOG group and TrxA group in the level of lesion-ceutric AQP-4 expression showing up by immunohistochemistry (P<0.05). Percentages of CD4~+ CD25~+ T cells in MOG group and GPSCH group were (4.71±1.61) % and (1.44±0.65) %, respectively, both of which were significantly lower than those in the normal control group or TrxA treated group (P<0.01). And the difference between MOG group and GPSCH group also reached statistics meaning (P<0.01). Normalized expression of Foxp3 mRNA in MOG group was 2.26± 1.97, and was not significantly higher than the 1.44±1.20 level in GPSCH group (P>0.05). However, they beth were statistically lower than that in the negative control group, namely 8.58±3.34 (P<0.01). Percentages of CD4~+ CD25~+ T cells was statistically correlated with expressions of Foxp3 mRNA (P< 0.05). Conclusion EAE induced in C57BL/6 mice with MOG~(Igd) is reproduceable. It shares the similar clinial signs and pathologic features with human multiple sclerosis(MS). Thus, we find a good way to further study the immune mechanisms of MS and also to search for the effective treatments.

Immunohistochemical Study on the SVCT Expression in the Spinal Cord of Experimental Allergic Encephalomyelitis (EAE) Model / 대한해부학회지

Experimental allergic encephalomyelitis (EAE) lesions by autoimmune inflammatory mechanism are characterized by the activation of microglia and astrocytes during the peak symptomatic stage of the disease. Besides it is well known that ROS and nitric oxide (NO), which is come out from activated inflammatory cells, play important role in the pathogenesis of EAE lesions. And vitamin C (L-ascorbic acid), which may protect from the deleterious effect by reducing iron (Fe2+) or copper (Cu2+) ions and maintain tissue homeostasis by removing of oxygen free radical, is inevitable to help many enzymatic reactions of cells. Previous report already investigated expression and functional analysis on various vitamin C transporters of vitamin C in many cell types. However, the researches for the vitamin C transporters are mostly performed in the normal state but not disease model yet. Therefore, for the first time, we investigated to know whether the SVCT1, 2 immunoreactivity may be observed in the astrocyte of EAE rat spinal cord. In the comparison of control and peak time group, the number of SVCT1, 2 immunoreactive cell was inclined to increase (P<0.05) as respectively 100+/-29.93, 135+/-34.62 in the control group, and 179+/-54.29, 349+/-73.56 in the peak time group. SVCT2 immunoreactivity was not doubly colocalized with GFAP antibody in the control group. In contrast, the astrocytes of the peak time group showed SVCT2 immunoreactivity in the perivascualr region and the cell number of doubly (SVCT2, GFAP) colocalized was 15+/-5.67 (P<0.05). We are firstly demonstrated that, in the evolving processes of EAE, astrocytes are able to use the vitamin C via the SVCT2. Taken all findings into consideration, the present data on the typical anti-oxidant vitamin C and its transporters, which may play a role in removing ROS, could be considered as a target to the therapeutic strategy of EAE and is also very useful to identify the characterization of vitamin C in the biological organism.

Establishment of and Effects of BPV on Experimental Allergic Encephalomyelitis in Lewis Rats / 医学研究杂志

Objective To establish the animal model of experimental allergic encephalomyelitis(EAE)in Lewis rats.Methods The EAE was induced by giving hypodermal injections in feet with spinal cords homogenate of guinea pigs(GPSCH)and complete Freund's adjuvant(CFA)containing 8mg.ml-1 bacille calmette guerin(BCG).The EAE model was evaluated by clinical manifestations and pathologic findings which was studied with the aid of light and electron microscope.Results The EAE in Lewis rats had an incidence of 9/10,a latency of 12.6?1.01d.Bordetella pertussis vaccine(BPV)could increase the morbidity,shorten the latency(P

Effects of prednisone on experimental allergic encephalomyelitis rats / 基础医学与临床

Objective To investigate the effects of prednisone(PRD) on experimental allergic encephalomyelitis(EAE) and the mechanism.Methods Lewis rats were injected with mixture of guinea pig spinal extracts and Freund's adjuvant(FA) to induce EAE.PRD(5 mg/kg) was orally given ten days after the injection.The clinical manifestation of rats,their peripheral blood lymphocytes CD3~(+),CD4~(+),CD8~(+) and NK subsets,and pathology changes of brain and spinal were studied on day 26.Results In model CD3~(+) ratio is 63.2%,CD4~(+)ratio 46.8%,CD8~(+)ratio 17.4% and NK ratio 10.2%.In PRD CD3~(+) ratio is 69.5%,CD4~(+)ratio 51.8%,CD8~(+)ratio 18.1% and NK ratio 5.3%.PRD obviously decreased the clinical scores of EAE rats and peripheral blood NK ratio,increased CD3~(+) and CD4~(+)ratio,inhibited brain inflammatory reaction and demyelination.However,PRD didn't affect peripheral blood lymphocytes CD8~(+) and CD4~(+)/CD8~(+)ratio.Conclusions PRD is effective in the treatment of EAE rats,and the therapeutic mechanism may be contributed to the decrease of the peripheral blood NK ratio and increase CD3~(+) and CD4~(+)ratio.

Changes of Nogo-A Expression in Spinal Cord of Rats with Experimental Allergic Encephalomyelitis / 实用儿科临床杂志

Objective To observe the time interval changes of the expression of Nogo-A in the intumescentia lumbalis of rats with experimental allergic encephalomyelitis(EAE),and explore the function of Nogo-A in the happening and development of EAE.Methods One hundred and six Wistar rats(female) were randomly divided into 2 groups:EAE group(70 rats)and control group(36 rats).Female Wistar rats were used to make EAE animal models,immunized by fresh cavia cobaya spinal cord homogenate and complete Freund adjuvant(CFA),at the same time,their blood brain barrier permeability was lifted by using pertussis stock solution.The rats those were coincident with the standard of being picked up were divided into 6 groups randomly according to the time of falling ill.In control group,fresh guinea pig spinal cord homogenate was replaced by sodium chloride,and rats were also divided into 6 groups randomly.Immunohistochemistry SP was used to detect expression of Nogo-A.Amyelination was detected by improved acid fuchsin and azulin myelin staining.Results In the control group,differences of Nogo-A expression in the intumescentia lumbalis were not obvious in each time group.In the EAE group,expression of Nogo-A could be observed in each time group,but their quantities were very different in contrast with those in the control group.One day after the rats had clinic symptoms,the expression of Nogo-A decreased to some extent;the third day,it reached the lowest point,and was significantly lower than that in control group(P

The effect and the significance of the matrix metalloproteinases in the pathological changes of the rat experimental allergic encephalomyelitis / 中国免疫学杂志

Objective:To investigate pathological changes and the expression of the matrix metalloproteinases in rat EAE.Methods:The pathological changes of EAE were studied in Wistar rat with the aid of light and electron microscope and the expression and distribution of MMP-2,MMP-9 in different tissues were detected with the method of immunohistochemistry.Results:Light microscopy showed inflammatory cuff around small blood vessels were evident, disintegration and disappear of myelin sheath were observed. Electron microscopy showed a lot of loose and fragmental spiremes of myelin sheath, the component of axons and nissl bodies in neurons were disappeared. MMP-2,-9 were expressed intensively in vascular endothelial cells, meninges and accumulative inflammatory cells.Conclusion:MMPs take the roles in every aspect of the pathological changes in EAE, it can destroy the blood brain-barrier, degrade the myeline sheath, damage the axons and generate immunogens.

Effects of neuropeptide Y on levels of serum interleukins in guinea pigs with experimental allergic encephalomyelitis / 临床神经病学杂志

Objective To observe the effect of neuropeptide Y(NPY) on the levels of serum interleukins(ILs)in guinea pigs with experimental allergic encephalomyelitis(EAE).Methods 30 guinea pigs were divided randomly into normol control group,EAE group and NPY group.The NPY was injected into the intracerebroventricular in NPY group one week before the EAE model be made.The EAE molders were made by injecting the homogenate of rat's spinal cord into the plantar of guinea pigs in groups EAE and NPY.Then,the neurological deficits score was observed everyday,the incidence and the delitescence of EAE were observed too.The levels of serum IL-1?,IL-2,IL-4,IL-6 and IL-8 were detected,and the neuropathological examination was used when symotoms peak period of EAE models.Results(1)The incidences of the EAE in groups EAE and NPY were 100% and 90%,the delitescences of EAE were(10.0?4.8)d and(25.4?12.6)d,the neurological deficits scores at the symotoms peak period were(3.60?0.52) and(1.80?1.14),respectively.There were significantly differences between the two groups(all P

The detection and significance of ICAM-1 and VCAM-1 in EAE rat brain / 重庆医科大学学报

Objective:The experimental allergic encephalomyelitis(EAE) is the most suitable animal model for investigation in the pathogenesis and the treatment of EAE.The expressions of ICAM-1,VCAM-1 in Wistar rat brain of acute EAE were investigated to study the mechanism and significance of their expressions in EAE rat brain.Methods:All experimental Wistar rats were inoculated intradermally in feet with emulsion contained guinea pig spinal cord homogenate and complete Freund’s adjuvant.The clinical manifestation and the pathological changes were observed with the aid of light and electron microscope to check the successful establishment of the model.The expressions of ICAM-1 and VCAM-1 were detected by the method of immunohistochemisty.The results of their expressions were compared by T test with controls.Results:Significant difference was found between the experimental group and the controul group in the expressions of ICAM-1 and VCAM-1 ( P

The establishment of multiple clinical processes on experimental allergic encephalomyelitis model of Wistar rat / 中国免疫学杂志

Objective:To investigate establishing the multiple clinical processes model and pathological characteristics of EAE model,and to provide the experimental value for reseaching MS. Methods:The animal model was established in Wistar rat by immunization with complete Freund adjuvant and GPSCH, the pathological changes of EAE were studied with the aid of light microscopy. Results : We can divide the EAE into five types by pathological changes and clinical manifestation: acute EAE, relapsing-remitting EAE, persistent progress EAE,benigh form EAE and delitescence EAE. Every type mainly showed congestion and inflammatory cuff of small blood vessels,disintegration and disappear of myelin sheath and degeneration of neurons,especially in spinal cord. Conclusion:Multiple clinical processes on EAE model of Wistar rat were established for the first time.it is an ideal animal model in studying MS.

Research on demyelinating mechanism in acute experimental allergic encephalomyelitis in cynomolgus monkeys / 中国免疫学杂志

Objective:To study demyelinating mechanism of the central nervous system in acute experimental allergic encephalomyelitis(EAE).Methods:EAE in cynomolgus monkeys was induced successfully by homologous brain white matter homogenate. Lymphocyte subset in blood and cerebrospinal fluid was monitored by flow cytometer. Pathological changes in brains of acute EAE monkeys were investigated by immunohistochemistry and electron microscopy. Results:In the CSF of acute EAE, CD4 + lymphocytes increased significantly, and CD8 + lymphocytes and B lymphocytes increased slightly whereas the control was normal. A lot of CD4 + lymphocytes and a few CD8 + lymphocytes infiltrated into temporal deep white matter of acute EAE, whereas the control was normal. Inner laminae of myelin sheath were loose and axon was separated, whereas outer laminae were normal. Although axon was preserved well, oligodendrocyte had a severe edema in cytoplasm with mitochondria swollen, crista blurred or broken, and nucleus lysised partly.Conclusion:It is oligodendrocyte rather than myelin sheath itself which is firstly attacked in the demyelination in EAE.

Role of the Cell-Mediated Immunity in Onset of EAE / 中国免疫学杂志

In the present paper, it is reported that immunelogic event during unset of experimehtaleallrgic encephalomyelitis (EAE) is studied by sevaeral immunological parameters, including thymectomy at birth in Guinea Pigs, identification of various cellular and humoral immunolo-gic functions. It is observed that thymectomy at birth suppressed onset of EAE and severityof EAE is related to cellular immunologic function but not to humoral immunity. All ofthese results indicated that cell-mediated immunity is responsible for the onset of EAE.