Læknablaðið : fylgirit - 01.05.1978, Side 134

Læknablaðið : fylgirit - 01.05.1978, Side 134
E. Genth, R. Kurze, W. Hartl Rheumaforschungsinstitut der Rheumaklinik Landesbad, Aachen FRG DEPRESSED ACTIVATION OF PERIPHERAL BLOOD LYMPHO CYTES BY MITOGENS IN RHEUMATOID ARTHRITIS There are conflicting reports about mitogen- induced stimulation of peripheral blood lymphocytes in patients with rheumatoid arthritis. LANCE et al and MENARD et al found lower lymphocyte stimulation rates in rheumatoid arthritis than in normals in contrast to WAXMAN and coworkers, who found no alterations. We studied the response of peripheral blood lymphocytes (LC) to Phythaemagglutinin-P (PHA-P), Concanavalin A (Con A) and Pokeweed mitogen (PWM) in 53 patients with rheumatoid arthritis (RA) and 14 healthy blood donors. The clinical data are listed up in the first slide. All patients suffered from classical or definite rheumatoid arthritis. Tests for rheumatoid factors were positive in 33 and for antinuclear antibodies in 23 cases. All except 4 patients were treated with antirheumatic drugs. Patients with AcetylsaUcylic acid therapy were exclused CLINICAL DATA OF PATIENTS STUDIED RHEUMATOID ARTHRITIS n=53 (ARA DEFINITE ANO CLASSICAL) SEX MALE 24 FEMALE 29 MEANAGE(YEARS) 49,4 »8 53*12,7 STEINBROCKER II III IV 'NDEX „= 2Q 23 8 2 RHEUMATOID FACTOR POSITIVE n= 33 ANA POSITIVE n= 23 DRUG THERAPY: GLUCOCORTICOSTEROIDS n = 9 D-PENICILLAMINE n = 9 AZATHIOPRINE n= 4 GOLD SALTS n = 3 ■ INDOMETHACIN n = 29 OTHER ANTIRHEUMATIC DRUGS n = 14 UNTREATED n = 4 BLOOD DONORS n = 14 SEX M/F 6/8 MEAN AGE (YEARS) 34,3 t11,0 Slide 1 . from the study, because there are several reports about a depressive effect of Acetyl- saUcylic acid on lymphocyte activation by mitogens. M eth ods: Triplicate minicultures of 300. 000 viable peripheral blood lymphocytes per well were set up with RPMI 1640 containing 4 mMol glutamine per ml, 200 IE penicillin 200 mg streptomycin and 207o of serum. LC were stimulated with 20 ul of mitogen in optimal and suboptimal concentration according to the doseresponse curve of the mitogen respective. The cells were incubated in a water vapour saturated atmosphere of 97o C02 and 937o air at 37°C for 72 hours. 22 hours before harvesting the cells on glass fibre filters the lymphocytes were pulsed with 1 uCi of tritium labeled thymidine. The radioactivity of the washed filter disks was counted in a liquid scintiUation counter. The results of tritium labeled thymidine uptake were expressed as average counts per minute of triplicate culturs. R esults: In the first experiment peripheral blood lymphocytes of 35 patients with rheumatoid arthritis and 10 healthy blood donors were culti- vated in autologous and pooled normal serum and stimulated with PHA-P, Con A and PWM in optimal and suboptimal dose. 3 or 4 patients and one blood donor were tested per day. The results of 10 randomly selected patients with RA and 10 controls are shown in the next slide. Statistical evaluation was performed with a split-split-plot design separately for each mitogen because of interactions between the source of lymphocytes i.e. patients and blood donors and the different mitogens. (2. Slide). In patients with rheumatoid arthritis lymphocyte activation with all three mitogens was signifi- cantly reduced independently from the mitogen concentration. Spontaneous tritium labeled thymidine uptake did not differ from the controls. The use of autologous or pooled normal serum yielded comparable results. The results of the 25 remaining patients were used to clarify the 132
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