Myasthenia gravis is one of the typical organ specific autoimmune disease and the CD5+ B-lymphocytes are known to be associated with the secretion of autoimmune antibodies. The authors performed the study to establish an animal model of experimental autoimmune myasthenia gravis (EAMG) by immunizing the nicotinic acetylcholine receptor (AChR) and to understand CD5+ B-lymphocyte changes in peripheral blood of EAMGs. Lewis rats weighing 150-200 g were injected subcutaneously three times with 50 microg AChR purified from the electric organ of Torpedo marmorata and Freund's adjuvant. The EAMG induction was assessed by evaluating clinical manifestations. The CD5+ B-lymphocyte was double stained using monoclonal PE conjugated anti-CD5+ and FITC conjugated anti-rat CD45R antibodies and calculated using a fluorescence-activated cell sorter (FACS). In three out of ten Lewis rats injected with purified AChR, the EAMG models were established. The animals showed definite clinical weakness responded to neostigmine; they had difficulty in climbing the slope, or easily fell down from a vertical cage. The range of CD5+ B-lymphocytes of peripheral blood in the EAMG models was 10.2%-17.5%, which was higher than in controls. In conclusion, the EAMG models were successfully established and the CD5+ B-lymphocyte expression in peripheral blood increased in EAMGs. This provided indirect evidence of the autoimmune pathomechanism of human myasthenia gravis.
Animal ; Antigens, CD5/immunology* ; B-Lymphocytes/immunology* ; Disease Models, Animal ; Myasthenia Gravis/immunology* ; Myasthenia Gravis/chemically induced ; Rats ; Rats, Inbred Lew

Myasthenia gravis and pemphigus are both considered to have an autoimmune basis. Although immunological and clinical studies have been performed on large numbers of patients with myasthenia gravis, the coexistence of myasthenia gravis and pemphigus foliaceus has rarely been described. We recently have the opportunity to study a 33-year-old female patient having both of these autoimmune diseases confirmed by various diagnostic methods. This rare coexistence of myasthenia gravis and pemphigus foliaceus has not been previously documented in Korea.
Adult ; Case Report ; Female ; Human ; Myasthenia Gravis/*complications/immunology/pathology ; Pemphigus/*complications/immunology/pathology

OBJECTIVETo explore the clinical significance of serum anti-ryanodine receptor (RyR) antibody in the diagnosis of myasthenia gravis (MG).

METHODSThe crude sarcoplasmic reticulum was prepared from rabbit skeletal muscle, and then purified by differential centrifugation to produce the antigen. The serum anti-RyR antibody levels in 74 patients with MG (including 21 patients with comorbidic thymomas) were determined with ELISA.

RESULTSWestern blot demonstrated the presence of RyR in purified crude sarcoplasmic reticulum. The positive rate of anti-RyR antibody was significantly higher in MG patients who had comorbidic thymoma compared with those who had no such comorbidity (P < 0.01). Also, the positive rate was closely correlated with the severity of MG.

CONCLUSIONSerum anti-RyR antibody test is helpful in the diagnosis of MG associated with thymoma and can be used to judge the outcome of MG.

Autoantibodies ; blood ; Humans ; Myasthenia Gravis ; blood ; complications ; immunology ; Ryanodine Receptor Calcium Release Channel ; immunology ; Thymoma ; complications ; Thymus Neoplasms ; complications

OBJECTIVETo observe the efficacy of Yiqi Qushi Recipe (YQR) in treating myasthenia gravis (MG) patients and its effects on their immune functions.

METHODSRecruited were 40 type I and II MG patients from clinics and wards of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from January 2009 to June 2011. They were randomly assigned to the treatment group (20 cases) and the control group (20 cases). Patients in the treatment group took YQR, one dose daily, while those in the control group took pyridostigmine 60 mg, three times a day. The therapeutic course consisted of eight weeks. The clinical efficacy, immunization indicators before and after treatment were observed. Meanwhile, the safety evaluation was performed.

RESULTSThe cured and effective rate was 75% and the total effective rate was 95% in the treatment group. They were 45% and 85% in the control group. Better results were obtained in the treatment group. Compared with the same group before treatment, IgA, IgG, and CD8 increased, IgM, CD4, and CD4/CD8 decreased in the treatment group, showing statistical difference (P < 0.05). There was no obvious change in each index of the control group after treatment (P > 0.05). Compared with the control group after treatment, IgA, IgG, and CD8 increased, CD4 and CD4/CD8 decreased in the treatment group, showing statistical difference (P < 0.05). During the course of treatment, mild diarrhea, nausea, and vomit occurred in two patients of the control group, while no adverse reaction occurred in those of the treatment group.

CONCLUSIONYQR could significantly improve clinical symptoms of MG patients, regulate their immune functions, with no obvious adverse reaction.

Adult ; CD4-CD8 Ratio ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Male ; Middle Aged ; Myasthenia Gravis ; drug therapy ; immunology ; Phytotherapy ; Young Adult

OBJECTIVEYoung C57BL/6 (B6) mice were treated with a specific tolerogen-dual analogue (Lys262-Ala207) intranasally to observe its effect on the invasion process of mice model and the clinical symptoms, to assess its clinical effects, and to explore the underlying mechanisms and feasibility of nasal mucosal tolerance explored.

METHODSPassively transferred myasthenia gravis (PTMG) was induced by mAb35 on B6 young female mice. Sixty mice were divided equally into three groups: tolerance group, model group and control group. Lys262-Ala207 was given intranasally (250 microg/mouse) to tolerance group with mAb35 for 10 successive days before immunization. Model group received PBS 50 microl only. The body weight and clinical scores were evaluated. The serum levels of AChRAb and the main cytokines (IL-4, IFN-gamma, TGF-beta1) were detected with ELISA.

RESULTSThe model group had typical myasthenia symptoms. B6 mice of tolerance group had less severe symptoms compared with control groups. The clinical symptoms of tolerance group were relieved. The level of AChRAb in tolerance group [(16.01 +/- 1.09) mg/L] was significantly lower than that of model group [(28.12 +/- 1.28) mg/L] (t = 44.37, P < 0.01). IL-4 and IFN-gamma levels in tolerance group [(141.02 +/- 3.11) ng/L, (187.99 +/- 4.67) ng/L] were significantly lower than those of model group [(193.37 +/- 3.95) ng/L, (320.46 +/- 2.14) ng/L] (t = 37.20, 51.69, P < 0.01). The level of TGF-beta1 in tolerance group [(437.19 +/- 1.93) ng/L] was higher than that of model group [(175.63 +/- 3.12) ng/L] (t = 36.07, P < 0.01). But there were still significant change as compared to those in control group (t = 26.65, 31.05, 49.02, P < 0.01).

CONCLUSIONSNasal administration of Lys262-Ala207 ameliorated muscular weakness in PTMG young mice. The therapeutic effect is possibly correlated with the function of immune system.

Animals ; Female ; Immune Tolerance ; Immunity, Mucosal ; Interleukin-4 ; blood ; Mice ; Mice, Inbred C57BL ; Myasthenia Gravis ; blood ; immunology ; Nasal Mucosa ; immunology ; Transforming Growth Factor beta1 ; blood

BACKGROUNDAntibodies against acetylcholine receptor, acetylcholinesterase, ryanodine receptor and titin have been found in patients with myasthenia gravis. However, the relations between these antibodies and character of myasthenia gravis are unknown. This study aimed to detect multiple antibodies in myasthenia gravis and to investigate its clinical significance.

METHODSThese antibodies were detected by enzyme-linked immunoabsorbent assay in 89 cases of myasthenia gravis, 66 cases of other neurological diseases and 66 healthy controls. The incidences of antibodies were compared using the chi-square test.

RESULTSAcetylcholine receptor, acetylcholinesterase, titin and ryanodine receptor antibodies were detected in 53.9%, 20.2%, 64.0% and 55.0% of myasthenia gravis patients respectively, higher than in patients of other neurological diseases and controls groups. The combination of the four antibodies assays provided 94.4% sensitivity and 84.0% specificity for the diagnosis of myasthenia gravis. Acetylcholinesterase antibody occurred more frequently in acetylcholine receptor antibody negative patients with adverse reactions to neostigmine test. Titin antibody provided 82.1% sensitivity and 52.5% specificity for myasthenia gravis with thymoma. Incidences of titin and of ryanodine receptor antibody were higher in late onset myasthenia gravis than in early onset myasthenia gravis. The proportion of titin antibody positive patients increased with the severity of myasthenia gravis as graded by a modified Osserman scale.

CONCLUSIONSTesting for acetylcholine receptor, acetylcholinesterase, titin and ryanodine receptor antibodies can offer a better diagnostic method for myasthenia gravis than each antibody test alone. Titin antibody combined with computed tomography was better for the diagnosis of thymoma. Titin antibody occurred most frequently in severe myasthenia gravis.

Adult ; Age of Onset ; Antibodies ; metabolism ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; immunology ; metabolism ; pathology ; Thymoma ; immunology

OBJECTIVETo study the clinical significance of serum anti-titin antibody level in the diagnosis of myasthenia gravis (MG) with thymoma.

METHODThe serum anti-titin antibody was analysed by ELISA method in 40 cases of health control group, 90 cases of MG, 17 cases of MG with thymoma and 7 cases of no-MG thymoma. The positive rate was compared among these groups.

RESULTSThe positive rate of anti-titin antibody was significantly higher in MG with thymoma patients than MG patients (94% and 3%, P < 0.01). According to the Osserman's classification, anti-titin antibody was present mostly in patients (43%) in IV stage, and also present in 2 cases of 7 who with no-MG thymoma.

CONCLUSIONSerum anti-titin antibody test is helpful in the diagnosis of MG with thymoma.

Adolescent ; Adult ; Aged ; Antibodies ; blood ; Connectin ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Muscle Proteins ; immunology ; Myasthenia Gravis ; complications ; diagnosis ; immunology ; Protein Kinases ; immunology ; Thymoma ; complications ; diagnosis ; immunology ; Thymus Neoplasms ; complications ; diagnosis ; immunology

OBJECTIVETo assess the therapeutic effect, timing of administration, complication prevention and management of the double filtration plasmapheresis (DFP) in the treatment of refractory myasthenia gravis (MG).

METHODSThirty-one patients with refractory MG were treated with KM 8800 membrane plasmapheresis monitor. DFP was performed every 3 days and the exchanging liquid was composed of 50 ml of 20% albumen and 1000 ml plasma substitute. Physical examination for absolute clinical score and blood sample was collected for AchR-Ab determination early in the morning on days 0, 3, 7, 14 of DFP.

RESULTSWith a total effective rate of 91.9%, complete recovery, basic recovery, improvement, and response was achieved in 2, 4, 11, and 17 patients, respectively, whereas the other 3 failed to respond. Hypotension occurred twice in 2 cases and was corrected after symptomatic treatment.

CONCLUSIONDFP may effectively lower blood AchR-Ab level of with minimal complications, and can be valuable for treatment of refractory MG.

Adolescent ; Adult ; Autoantibodies ; blood ; Child ; Female ; Filtration ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; blood ; therapy ; Plasma Exchange ; methods ; Receptors, Cholinergic ; immunology ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo observe the therapeutic effect of electroacupuncture (EA) warming therapy on myasthenia gravis (MG) and effect on IL-4 in the patient.

METHODSSixty patients with MG were randomly divided into two groups, 30 patients in each group. The observation group were treated with EA warming therapy with Tanzhong (CV 17), Shimen (CV 5), Guanyuan (CV 4), Zhongwan (CV 12), Yanglingquan (GB 34) selected as main points, and oral administration of Pyridostigmine (90 - 240 mg each day) and Prednisone (30 - 60 mg each day). The control group were treated with oral administration of Pyridostigmine (240-480 mg each day) and Prednisone (60-100 mg each day). Clinical therapeutic effects and serum Interleukin-4 (IL-4) levels before and after treatment were observed.

RESULTSThe total effective rate was 93.3% in the observation group, which was better than 70.0% in the control group (P < 0.01); after treatment, the serum IL-4 levels in the two groups significantly decreased (P < 0.01), the decrease of IL-4 in the observation group being significantly better than the control group (P < 0.05).

CONCLUSIONEA warming therapy combined with western medicine has a significant therapeutic effect on myasthenia gravis. One of the mechanisms possibly is to restrain specific immune reaction by regulating the level of IL-4.

Acupuncture Points ; Adult ; Combined Modality Therapy ; Electroacupuncture ; methods ; Female ; Humans ; Interleukin-4 ; blood ; Male ; Middle Aged ; Myasthenia Gravis ; immunology ; therapy ; Prednisone ; administration & dosage ; Pyridostigmine Bromide ; administration & dosage

OBJECTIVE: To explore the role of IL-21 in the pathogenesis of myasthenia gravis (MG) and its influence on the the class switch of anti-AChR antibodies. METHODS: Blood was taken from 26 patients and 18 healthy controls, and the expression of IL-21R mRNA in peripheral blood mononuclear cells (PBMCs) was detected by RT-PCR. The expression of IL-21R on B lymphocytes was measured by flow cytometry, while the concentrations of serum IL-21 and the levels of anti-AChR-IgG and its isotype IgG(1), IgG(2), and IgG(3) were tested by ELISA. RESULTS: The serum concentration of IL-21 in the MG group was higher than that in the control group (31.686±8.499 pg/mL, 15.147±6.366 pg/mL) and the difference was significant (P<0.01). IL-21R mRNA expressed on PBMCs in the MG group was higher than that in the control group (0.139±0.052, 0.101±0.022), and the difference was significant (P<0.05). There was no difference between ocular MG and generalized MG subgroup (P>0.05). Compared with the control group, the expression of IL-21R on B lymphocytes also increased in the MG group (P<0.05). In the anti-AChR-Ab positive MG group, the serum concentration of IL-21 showed positive correlation with anti-AChR-IgG(P<0.05),but no correlation with its isotype IgG(1), IgG(2), and IgG(3), respectively(P>0.05). Expression of IL-21R mRNA in the PBMCs showed no correlation with the level of serum anti-AChR-IgG and its isotype IgG(1), IgG(2), and IgG(3), respectively(P>0.05); however the expression of IL-21R in B lymphocytes showed positive correlation with anti-AChR-IgG and it's isotype IgG(1) and IgG(3) (P<0.05,P<0.01,P<0.05), but no correlation with IgG(2) (P>0.05). CONCLUSION IL-21 might induce the class switch of anti-AChR antibodies to IgG(1) and IgG(3) isotype through IL-21R on B lymphocytes which promotes the pathogenesis of the MG.
Adolescent ; Adult ; Autoantibodies ; classification ; immunology ; Female ; Humans ; Immunoglobulin Class Switching ; immunology ; Immunoglobulin G ; classification ; Interleukins ; blood ; genetics ; Male ; Middle Aged ; Myasthenia Gravis ; blood ; immunology ; RNA, Messenger ; blood ; genetics ; Receptors, Cholinergic ; immunology ; Receptors, Interleukin-21 ; blood ; genetics ; Young Adult