Læknablaðið : fylgirit - 01.05.1978, Page 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
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