BACKGROUND: A human B-cell subpopulation which is identifiable by the expression of cell surface antigen Leu-1 (CD5) is known to be responsive for the antoantibody secretion. The exprimental autoimmune myasthenia gravis(EAMG) could be induced in animals by injecting AChR from electric eel and had the same clinical and pathophysiological characteristics with human myasthenia gravis(MG). The authors performed the study to compare the frequncy of CD5 positive B-lymphocytes in EAMGs with those in human MGs and to understand whether the EAMG showed the similar immunological feature as in human MG. METHOD: For EAMG the 50ug AChR from Torpedo Marmorata with Freund's adjuvant were injected to Lewis rats of 150-200mg three times. The CD5 positive B-lymphocytes from peripheral blood were double stained by the monoclonal PE conjugated anti-CD5 and FITC conjugated anti-rat CD45 antibodies in the EAMGs and by the PE conjugated ani-Leu-11(CD5) and FITC conjugated anti-human Leu-12(CD19) antibodies in human MGs. The expression of positive CD5 positive B-lymphocytes were calculated by the fluorescent activated cell sorter(FACS). RESULTS: The mean CD5 positive B-lymphocytes expression in four EAMGs was 15.05% with ranging from 10.2% to 20,0%, which was increased compared with those in control rats. The mean frequency of CD5 positive B-lymphocytes were 25.2+/-15.05% in human MG(N=25) and 16.0+/-13.5% in normal controls (N=20) respectively, which did not show any significant difference (P=0.08). However the expression of CD5 positive B-lymphocytes in human MGs was significantly correlated with the titer of anti-AChR antibody (P=0.04). CONCLUSION: The increased expression of CD5 positive B-lymphocytes might be associated with the EAMG pathomechansms, but their expression only showed possible correlation with the anti-AchR antibody titer with minimal role in pathogenetic process of human MG. Therefore it would be suggested that immunological process of EAMG (acute form of MG) be a little different from that of human MG, chronic form.
Animals ; Antibodies ; Antigens, Surface ; B-Lymphocytes* ; Electrophorus ; Fluorescein-5-isothiocyanate ; Freund's Adjuvant ; Humans* ; Myasthenia Gravis* ; Rats ; Torpedo

BACKGROUND: The aim of study is to investigate the initial functional changes of muscle in rats induced to have myasthenia gravis, experimental autoimmune myasthenia gravis (EAMG). The authors investigated the functional changes of muscle evaluated by mechanomyography (MMG) and the expression of acetylcholine receptors (AChRs). METHODS: After the institutional approval, 39 male Lewis rats were randomly allocated into study. 26 animals were immunized to induce EAMG by Torpedo AChR (T-AChR) emulsified with complete Freund's adjuvant (CFA) and phosphate buffer saline (PBS)/bovine serum albumin (BSA) 0.01% at the base of tail, and received booster immunizations twice by T-AChR with incomplete Freund's adjuvant (IFA) and PBS/BSA 0.01% at all different site on the upper back. 13 animals were sham immunized as control group by the same method of EAMG except T-AChR. Clinical EAMG scores were examined. Anti T-AChR and anti rat-AChR (R-AChR) antibodies (Ab) were compared by using (125)I-alpha-bungarotoxin ((125)I-alpha-BuTx) radioimmunoassay. Under the anesthesia, neuromuscular functions were monitored by MMG using single twitch (ST) and TOF. AChRs were quantitated using (125)I-alpha-BuTx. RESULTS: Overall weight gain and final body mass, muscle force (ST), specific muscle force of ST, TOF fade ratio and AChRs were reduced in EAMG score 3 compared to control (P < 0.0001). Anti T-AChR Ab and anti R-AChR Ab were increased in score 3 EAMG (P < 0.0001). CONCLUSIONS: EAMG score 3 rats showed characteristic neuromuscular functions as depressed initial ST and its specific force, initial TOF fade and increased anti AChR Abs. Those above characteristics had significant correlations with the clinical EAMG scores. AChRs were significantly down-regulated according to their functional characteristics and clinical EAMG scores.
Acetylcholine* ; Anesthesia ; Animals ; Antibodies ; Freund's Adjuvant ; Humans ; Immunization, Secondary ; Male ; Myasthenia Gravis* ; Myasthenia Gravis, Autoimmune, Experimental ; Radioimmunoassay ; Rats* ; Receptors, Cholinergic* ; Serum Albumin ; Tail ; Torpedo ; Weight Gain

Duchenne Muscular dystrophy(DMD) is a debilitating X-linked muscle disease and dyskophin is a muscle membrane protein, which is recently discovered through reverse genetics by Kunkel et al(1987). We evaluate the dystrophin distribution by irnmunohistochemical staining with anti Torpedo dyskophin antibody in the muscle biopsy materials from 11 clinically and pathologically diagnosed Duchenne type muscular dyskophy and 23 controls of other neuromuscular disorders or normal amputed legs. Normal staining of dystrophin were found in all the muscle preparation from 23 controls. In 10 of 11 pahents with Duchenne type muscular dyskophy diagnosed clinically and pathologically, reaction to anti-Torpedo dystrophin antibody was absent or markedly deficient. However, in one subject with definite DMD clinically, the immunostaining showed normal dense staining. He was a 5-year-old boy who was presented with abdominal pain and general muscle weakness, and his final diagnosis were choledocal cyst and Duchenne muscular dyskophy. Therefore it could be concluded that the immunohistochemical staining with anti-Torpedo dystrophin antibody should prove helpful in delineatior of myopathies that overlap clinically with Duchenne type progressive muscular dyskophies and it shows prornise as an accurate tool for the diagnosis of DMD and for the evaluation of therapeutic effects.
Abdominal Pain ; Biopsy ; Child, Preschool ; Diagnosis ; Dystrophin* ; Humans ; Leg ; Male ; Membrane Proteins ; Muscle Weakness ; Muscular Diseases ; Muscular Dystrophies* ; Reverse Genetics ; Torpedo