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In vitro peripheral blood and synovial fluid lymphocyte interactions.

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In Vitro Peripheral Blood and Synovial Fluid
Lymphocyte Interactions
Marie M. Griffiths and Ralph C. Williams, Jr.
In vitro interaction of autologous peripheral blood and synovial fluid
lymphocytes was studied using cells isolated from patients with various
forms of arthritis. Addition of mitomycin-treated peripheral blood lymphocytes from patients with inflammatory arthritis to in vitro cultures of autologous synovial fluid lymphocytes caused an increased incorporation of
tritiated thymidine by the synovial fluid lymphocytes. However when the
reverse situation was tested, no stimulation of peripheral blood lymphocytes by mitomycin-treated synovial cells was recorded. In some instances synovial fluid lymphocytes were observed to exert an apparent
cytotoxic or inhibitory effect on autologous peripheral blood lymphocytes. The latter response pattern was found to occur to a much greater
degree with lymphocyte preparations from patients with rheumatoid arthritis than in those with other forms of inflammatory joint disease.
Recently considerable attention has been intrinsic to the disease itself alter autoldirected at lymphocytes and their relation- ogous tissue in such a way that the body
ship to various connective tissue disorders is stimulated to mount an immune re(1-6) as well as to animal models of auto- sponse-both cellular and humoral-against
immune disease such as NZB mice (7-9). If such altered materials. Second if the ansensitized lymphocytes play an effector role tigens are concentrated in areas characterin diseases like rheumatoid arthritis, sev- istically affected by the disease, one would
eral possible mechanisms might be postu- then expect to find in the diseased joints
lated. First it is conceivable that processes or joint tissue relatively large amounts of
specific disease-related antigens which could
From the Arthritis Unit, Department of Medi- be used in an in vitro examination of the
cine, Bernalillo County Medical Center, University cellular immune reaction to such antigens.
of New Mexico School of Medicine, Albuquerque, The present study, conceived as a frontal
New Mexico, 87131.
Supported in part by grants AM 13824-03 and assault on this problem, centered around
TO1 A100343-03 from the US Public Health Service, observations of the in vitro response of
and in part by a grant from the Kroc Foundation. cultured peripheral blood lymphocytes to
RALPH c. WILLIAMS, JR, MD: Professor and Chairman, Department of Medicine, University of New the presence of autologous synovial fluid
Mexico School of Medicine, Albuquerque, New cells and vice versa. It was postulated that
Mexico, 87131; MARIE hi. CRIFFITHS, PhD: Depart- if synovial fluid or synovial fluid cells conment of Medicine, Division of Infectious Diseases,
University of Utah Medical Center, Salt Lake City, tained large amounts of rheumatoid-speUtah, 84112.
cific or neoantigens somehow produced by
Address reprint requests to Dr Williams.
the disease itself-then it was possible that
Submitted for publication June 22, 1973; accepted
soluble or cellular components of synovial
October 17, 1973.
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
111
GRlFFlTHS AND WILLIAMS, JR.
fluid might produce stimulation of autologous peripheral blood cells previously sensitized to such antigens during the course
of the disease.
Mitosis accompanying the blast transformation, which occurs when sensitized lymphocytes interact with a specific antigen,
can be measured as an increased incorporation of tritiated-thymidine into acid precipitable nuclear material above the background incorporation of lymphocytes cultured in the absence of added antigen. This
method was used in the experiments to be
reported.
Results presented here are of considerable interest since they indicate that cell to
cell interactions within individual patients
are probably subject to local controlling
factors which are not yet well understood
but may relate to T- and B-cell mediators
and possibly cell-to-cell intercourse.
cells. In two instances, specific preparations of
macrophages were made from mononuclear synovial
fluid cells using the albumin flotation method described by Bennett and Cohn (11).
Lymphocytes that were to be treated with mitomycin were suspended in HBSS-FC at 1 X 10' cells
per milliliter and incubated at 37°C for 30 minutes with Mitomycin-C (Sigma Chemical Company)
at 40 pg/ml. The cells were then washed three
times with HBSS-FC and resuspended in culture
medium.
Culture medium consisted of Minimum Essential
Medium (Eagle's salts) supplemented with 20%
heat-inactivated fetal calf serum, 2 mM glutamine,
and antibiotics (penicillin, 100 units/ml and streptomycin, 100 pglml). Lymphocyte cultures of 1.4
ml final volume were prepared in triplicate and
incubated at 37°C for 120 hours in a humidified
atmosphere of 574 CO, in air. One microcurie of
tritiated thymidine (New England Nuclear, 20
pCi/mmole) was added to each tube for the final
6 hours of incubation.
Cultures were terminated by the addition of 3
ml of cold PBS containing 5 X 1 P molar nonradioactive thymidine and centrifugation. The cell
pellet was washed once with PBS-thymidine and
stored at -20°C. The samples were then thawed
MATERIALS AND METHODS
and nuclear material precipitated with cold 10%
trichloroacetic acid (TCA). Each precipitate was
washed two times with 5% TCA, once with absolute methanol, dried, and solubilized with NCS
solubilizer (Amersham-Searle). Radioactivity was
measured by liquid scintillation counting techniques. Background incorporation of 1 X 108 normal lymphocytes under these culture conditions
raiiged from 3700 to 6100 CPM (mean average of
triplicate cultures zk 10%) after 5 days in culture.
Lymphocyte responsiveness to mitogen stimulation
was assessed in each experiment by the addition of
phytohaemagglutinin (PHA) a t a predetermined
optimum concentration to triplicate cultures. These
controls were stopped at 72 hours of incubation
after a 4-hour pulse with 1 pCi of tritiated thymidine. Under these conditions normal responses of
lymphocytes from healthy individuals were 170,000
to 250.000 CPM (& 10%) above an unstimulated incorporaiion of 2000 to 3000 CPM (* 10%). For all
experiments reported, PBL cultures demonstrated
a normal response to PHA. SFL cultures demonstrated a lower stimulation of a 15- to 40-fold
increase above background at the two-cell concentrations used in the comparative cultures. All data
All patients studied were followed in thc Arthritis Clinic, Bernalillo County Medical Center,
Albuquerque, New Mexico. Blood samples were
obtained from patients undergoing joint aspiration
for clinical reasons. Both blood and synovial fluid
samples were collected in heparinized syringes.
Synovial fluid samples were treated at 37°C with
hyaluronidase (Calbiochem, 523 NF units/mg) at
25 pg/ml for 30 minutes. Lymphocytes were then
collected simultaneously from the two samples by
the Ficoll-Hypaque gradient centrifugation method
essentially as described by Boyum (lo), washed
twice with Hank's Balanced Salt Solution containing 5 % heat-inactivated fetal calf serum (HBSS-FC)
and resuspended in culture medium.
Lymphocyte preparations were examined using
Wright's stained smears. Differential counting of
peripheral blood lymphocyte preparations (PBL)
indicated an average of 95y0 small or medium sized
lymphocytes and 5% monocytes. In the case of
synovial fluid lymphocyte preparations (SFL) the
majority (8070-900/,)of cells appeared to be medium or large lymphocytes and the remainder
monocytes, macrophages, and polymorphonuclear
112
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
LYMPHOCYTE INTERACTIONS
Table 1. Mixed Cell Cultures Using Synovial Fluid Cell Pellets
and Autologous Peripheral Blood Lymphocytes
Background CPM
Patient
EC
MG
Diagnosis
Classic RA
Pseudogout
PBL'
207 (+53)
1,540 (+153)
SFCt
5 (f4)
680 ( t 1 7 )
Mitogen Response
PBL
+ PHA
36,352 (*1,811)
221,381 ('28,381)
Mixed Cell Cultures
(PBL SFL)
+
Calc CPM Actual CPM
212
2,220
667(+62)
1,724 ( + 9 l )
'Peripheral blood lymphocytes prepared by Ficoll gradient centrifugation were cultured at 1 X 10'
cells per 1.2 ml of medium.
tSynovial fluid cells (SFC) represent the total cell population of the synovial fluid obtained by centrifugation of the synovial fluid sample at 1,500 rpm for 15 minutes and then added to the assay
system such that the final cell concentration would be equivalent to that in the original synovial
fluid sample. Values are the mean CPM of triplicate cultures standard deviation.
*
trifugation of the synovial fluid instead of
the total synovial fluid cell population. The
results of 13 separate experiments are presented in Table 2. The values tabulated in
the first three columns of Table 2 represent
the background incorporation of tritiated
thymidine by PBL and synovial fluid lymphocytes (SFL) when cultured separately and
RESULTS
in the absence of any stimulants. T h e next
During attempts to demonstrate an in four columns compare the expected results
vitro mitogenic response of lymphocytes (calculated by addition of the individual
from patients with rheumatoid arthritis to background incorporations) with the actual
components of the patient's own synovial values obtained when the two cell populafluid, it was observed that addition of the tions were cultured together at the indicell pellet, obtained by low speed centri- cated cell concentrations. Of the seven
fugation of synovial fluid, to cultures of assays using cells from patients with rheuautologous peripheral blood lymphocytes matoid arthritis, three showed a significant
(PBL) resulted in an incorporation of tri- increase in actual thymidine incorporation
tiated thymidine which was significantly (patients 1,2,3) and in 3 cases (patients
higher than that expected to result from a 5,6,7) the effect was one of decreased insimple additive effect (Table 1, patient 1). corporation of tritiated thymidine. Mixed
This effect was not seen in a subsequent cultures of peripheral blood lymphocytes
experiment using peripheral blood lym- (PBL) and synovial fluid lymphocytes
phocytes and synovial fluid cells (SFC) from (SFL) from patients with other forms of
a patient with pseudogout (Table 1, pa- arthritis showed similar trends but the
tient 2).
changes were not definitive. These data are
Because of the interesting results noted also shown in Table 2 (patients 8-13).
in patient 1 (Table 1) this type of experiIn 2 cases (patient 5 (FMG) and patient
ment was repeated using lymphocyte-rich 11 (EA) attempts to demonstrate stimulafractions obtained from Ficoll-gradient cen- tion of peripheral blood lymphocytes by
are presented in the tables as mean cpm of triplistandard deviations from the mean.
cate cultures
Plasma and cell-free supernatants from synovial
fluid samples were assayed for cytotoxicity against
lymphocytes from a panel of 6 normal donors by
the microdroplet technique of Terasaki and McClelland (12) as modified by Lies et a1 (13).
*
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
113
P)
h)
1
Classic RA
Classic RA
Juvenile RA
Classic RA
Classic RA
Classic RA
Classic RA
Reiter's
syndrome
Reiter's
syndrome
Gout
Ankylosing
spondylitis
Pseudogout
Possible RA
MW
LW
MC
LW
FMG
CV
CV
WM
EA
743 ('61)
9,937 (21,654)
2,877 (a475)
5,785 (21,581)
18,670 (+5,920)
6,470 (+2,000)
4,126 (+1,011)
1,912 (k62)
30,146 ('4,200)
35,384 (+2,853)
1,091 (2171)
26,929 (*5,030)
24,992 (k1,960)
PBL (CPM)
(10' cells)
MLC
60 (+8)
22 (22)
259 (+17)
262 (250)
3,488 (+646)
87 (k22)
63 (220)
573 (+186)
1,152 (2570)
250 (268)
436 (k86)
14 (+2)
267 )
4
3
'
(
*
709 (2190)
-
130 (25)
-
1,272 (+279)
314 (285)
9,959
3,136
803
6,047
22,158
6,557
4,189
2,485
31,298
35,634
1,527
20 (26)
26,943
794 (--+209) 25,259
445 (k40)
2,372 (2120)
4,898 (2200)
632 (+65)
1,126 (+27)
7,049(+261)
6,043 (21,808)
5,964 (k791)
31,194 (*7,681)
6,530 (21,944)
7,905 (f2,395)
4,245 (2161)
52,273 (+2,978)
41,479 (3,605)
597 (+is)
17,260 (+3,200)
18,355 (22,900)
3,586
-
873
-
7,057
6,784
3,298 (+260)
-
1,240 (+39)
-
7,801 ('114)
6,200 (f943)
-
12,241 (k594)
12,358 ('1,327)
-
26,949
25,786
(+166)
(2558)
(+4,200)
(+2,806)
9,911
5,710
68,182
39,801
4,571
4,284
35,044
36,016
SFL (CPM)
PBL (10') -k SFL (2x106) PBL (10') -k SFL (4x106)
(2X1OScells) (4X106 cells) Calc CPM Actual CPM Calc C M Actual CPM
MLC
0
1+
ND
0
0
0
3+
3-k
1-k
0
2+
ND
0
0
3+
2+
0
0
0
ND
0
Synovial
Fluid
0'
0
0
ND
2+
Plasma
Cytotoxicity'
standard deviations.
Values are presented as mean CPM for triplicate cultures
'Cytotoxicity was scored as 1+ if samples showed between 10% and 25% average killing of test lymphocyte panel; 2-k for 25%-50%; 3-k
for 50%-75%; and 4+ for 75%-100% killing.
JG
JO
JM
PK
Diagnosis
Patient
Background Incorporation
Table 2. Incorporation of Tritiated Thymine Into Lymphocytes from Peripheral Blood and Synovial Fluid Separately and in Mixed Cultures
a
c
1
U
4
1
g
5
35,384 (*2,853)
35,384
24,992 (+1,960)
24,992
30,146(+4,200)
30,146
6,470(*2,000)
6,470
5,785(+1,581)
5,785
-
12
9
4
28
16
21
11
19
46
3
-
12
15
SFL,
-
~
36,942(*4,800)
32,575(+2,700)
27,880(*2,815)
23,195(+2,305)
32,138(*1,816)
46,584(+2,356)
7,430(*goo)
8,005(+1,005)
3,594(*1,169)
-
+ SFL,
3,783(&1,179)
2,997(*873)
PBL
MLC
435 (+49)
435
104 (216)
104
156 (215)
156
123 (*22)
123
237 (*153)
237
-
110 (+lo)
-
250 (*68)
632 (+65)
267 (+34)
794 (+209)
1,152(+570)
4,898(k200)
87 (222)
314 (k85)
162 (*50)
1,272(k279)
-
589 (+212)
-
SFL
PBL,
Background (CPM)
Mitomycin-treated PBL
6,303(k714)
8,288(t858)
4,395(+1,143)
4,306(*728)
8,019(k287)
16,570(+2,670)
733 (*112)
983 (*73)
1,082(+404)
4,219(+353)
-
SFL
2,468(f15)
-
PBL,
MLC
22
12.5
16
5.4
6.8
3.3
7
2.7
3
3
4
-
s11
*
PBL and SFL refer to peripheral blood lymphocytes and synovial fluid lymphocytes prepared and treated with mitomycin as described in
standard deviations.
Methods. Values are presented as mean CPM of triplicate cultures
*Peripheral blood leukocytes (PBL) were used at a concentration of 1 X 10' cells and synovial fluid cells (SFL) were tested at two concentrations designated as a = 2 X 10' and b = 4 X 109
tSI refers to stimulation index as calculated by the ratio of observed counts to baseline counts recorded for synovial fluid cells cultured
alone and represents increased growth of SFL in presence of mitomycin-treated PBL.
Reiter's
syndrome
Reiter's
syndrome
b
WM-a
b
PK -a
b
Classic RA
Classic RA
Classic RA
4,126(+l,Oll)
4,126
Classic RA
Juvenile RA
b
b
b
b
PBL'
Diagnosis
MC -a
CV -a
LW -a
EC -a
MW-a'
Patient
Background (CPM)
Mitomycin-treated SFL
Table 3. One-way Mixed Leucocyte Culture Results Using Autologous Peripheral Blood and Synovial Fluid Leukocytes
u)
5
z
2
rn
3
B
f
-I
3rn
0
0
5I
2
GRlFFlTHS AND WILLIAMS, JR.
graded numbers of purified macrophage
preparations were essentially negative. In
the studies recorded here addition of such
macrophages to autologous mixed cultures
did not appear to show a consistent stimulatory or depressive effect; however this
aspect of the problem was not intensively
investigated due to the restrictions of sample size.
Paired plasma and cell-free synovial fluid
samples were obtained at the time of lymphocyte preparation and stored at -2OOC.
T h e results of a survey of these samples for
lymphocytes against a panel of normal donor lymphocytes are presented in the righthand column of Table 2. It was of interest
that the four plasma samples, which demonstrated cytotoxic activity (samples 5,6,7,
12), correspond to the cases in which addition of synovial fluid lymphocytes to peripheral blood lymphocytes decreased the incorporation of tritiated thymidine.
These initial results indicated that in
many instances, particularly in the case of
individual subjects with rheumatoid arthritis, a n additional increment of cell proliferation was occurring when mononuclear
leukocytes (predominantly lymphocytes)
from peripheral blood were cultured together with a similar population of cells
from synovial fluid. However it was not
clear from such studies which of the two
cell populations was responsible for the increased thymidine incorporation. For this
reason, studies using a system of one-way
mixed leukocyte cultures and mitomycin
treatment of one cell population-either
the synovial fluid or peripheral blood lymphocytes (14)-were performed. These studies are summarized in Table 3.
T h e data in Table 3 demonstrate that in
each case studied, the increased thymidine
incorporation observed when synovial fluid
lymphocytes were cultured with peripheral
116
blood lymphocytes was due to increased
growth of the synovial fluid lymphocytes.
T h e addition of mitomycin-treated synovial
fluid lymphocytes (SFLm) to cultures of peripheral blood lymphocytes did not cause
any significant increase in incorporation of
thymidine above background. However,
when mitomycin-treated peripheral blood
lymphocytes (PBLm)were added to cultures
of synovial fluid lymphocytes a significant
stimulation was observed.
Under these circumstances, positive results (ie, stimulation of SFL by PBLm) did
not appear to be restricted to patients with
rheumatoid arthritis or to correspond with
the direction or extent of incorporation by
the mixed cultures as presented in Table 2.
Thus patient LW whose cells demonstrated
only a slight increase under the protocol
used to obtain the data in Table 2 (no. 4)
(ie, no mitomycin treatment of either cell
population) showed a ten- to twentyfold
stimulation of synovial fluid lymphocytes
by mitomycin-treated autologous peripheral blood lymphocytes under the assay conditions of Table 3 (no. 3). Of great interest
was the fact that in the same patient no
stimulation of peripheral blood lymphocytes occurred in the presence of mytomycin-treated synovial fluid cells. I n addition,
the two patients with Reiter’s syndrome,
who in Table 2 appeared to be negative
controls, demonstrated three- to sevenfold
stimulation of synovial fluid lymphocytes
by mitomycin-treated peripheral blood lymphocytes (Table 3, nos. 6 and 7); in like
manner reverse stimulation of peripheral
blood lymphocytes by mitomycin-treated
synovial fluid cells was not recorded.
I n one instance (Table 2, no. 7) where a
decreased incorporation of tritiated thymidine occurred in the mixed culture, it was
determined that these results were indeed
due to decreased incorporation by the pe-
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
LYMPHOCYTE INTERACTIONS
Table 4. Experiments Assaying the Importance
of Cell Density and Cell-to-Cell Interaction
in Peripheral Blood and Synovial Fluid
Lymphocyte Cultures
Cell Conditions
PBL (1 X 10')
SFL (4 x 105)
PBL (1 X 10')
SFL (4 X 10')
PBL (1.4 X 10')
+
tion and stimulation of the two cell populations was due to differences in viability
in culture.
CPM
24,992 (+2,857)
794 (k265)
12,358 (*1,655)
34,427 (+2,549)
Culture conditions were as described in the text.
PBL = Peripheral blood lymphocytes
SFL = Synovial fluid lymphocytes
Values are mean cpm of triplicate cultures
standard deviations.
*
ripheral blood lymphocytes and were attributable to the presence of SFL. These data
are presented in Table 4. It can be seen
that increasing the initial concentration of
peripheral blood lymphocytes to a level
equivalent to the combined concentration
of autologous PBL and SFL in the mixed
culture (LC 1.4 x lo6 total cells) resulted
in the expected additive growth effect.
Thus, the decrease observed in the mixed
cell culture could not be interpreted as
resulting from overgrowth of the higher
cell population during the incubation period, and must have been due either to cell
death or to inhibition of mitosis.
In 4 instances (Table 3, patients 4-7)
cell viability after 3 and 5 days in culture was assessed by trypan blue exclusion. From an initial cell density of 1 x 10"
cells per 1.4 ml culture volume, the percent
recovery of viable cells was comparable for
the two cell populations (SFL, 420/,-5370;
PBL, 47%-670/,). At the lower cell density
of 2 x 105 cells per 1.4 ml (Table 3, patient
7). the percent recovery of viable cells after
5 days was even higher (SFL, 80%; PBL,
95%). Values are mean averages of duplicate determinations on each culture. These
data would argue against the possibility
that the observed differences in incorpora-
DISCUSSION
When lymphocytes from synovial effusions of patients with rheumatoid arthritis
were cultured in vitro with autologous peripheral blood lymphocytes, the incorporation of tritiated thymidine into the mixed
cultures significantly deviated from the expected value calculated from separate and
simultaneous cultures of the two individual cell populations in 6 of 7 cases at one
or both of the two cell concentrations
tested. This type of response pattern was
not observed in comparable assays using
cells obtained from patients having inflammatory conditions of nonrheumatoid origin. One-way mixed lymphocyte cultures
demonstrated that the observed increases
in thymidine uptake were due primarily to
growth of the lymphocytes derived from
synovial fluid and not to the peripheral
blood lymphocytes. Under these conditions
however the results did not appear to be
specific for rheumatoid arthritis. Increased
blastogenesis of the synovial fluid lymphocytes could be interpreted as the response
of sensitized lymphocytes to an antigenic
component carried on the peripheral blood
lymphocytes. However, since any existing
disease-specific antigen would be expected
to be concentrated in the affected joints,
this is not the direction of stimulation we
expected to encounter. It is possible that
the peripheral blood lymphocytes are obtained from the patient in a highly activated state as a result of the disease process
itself. Although mitomycin treatment is
known to inhibit cell mitosis (16), it may
not inhibit release of preformed mediators.
Thus peripheral blood cells which were
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
117
GRlFFlTHS AND WILLIAMS, JR.
activated in vivo and prior to mitomycin
exposure could possibly, in culture, release
preexisting mitogenic factors which would
initiate blastogenesis of any added nonmitomycin-treated cell population, in this case
synovial fluid lymphocytes.
In four experiments using peripheral
blood lymphocytes from patients with rheumatoid arthritis (Table 2, patients 3,4,6,7)
the background incorporation of thymidine
was much higher than that routinely encountered using lymphocytes from healthy
individuals. In 3 cases (Table 2, patients 5
and 11; Table 3, patient 1) the background
incorporation of PBL was much lower than
expected. It should be noted that experiments 6 and 7 of Table 2 represent cells
from the same patient studied at a 4-week
interval at which time the initial high
background incorporation was repeated.
These results apparently reflect the disease
state at the time samples are obtained and
are compatible with the possible consideration that in certain cases the peripheral
blood lymphocytes are obtained in an activated state.
The blast transformation assay in and of
itself does not distinguish between inhibition of cell mitosis (16) and cell death. The
decrease in incorporation of tritiated thymidine (>50%) by the peripheral blood
lymphocytes upon culture with synovial
fluid lymphocytes which was observed in
three of the active rheumatoid cultures and
equivocally (SO’%) in only one of the nonrheumatoid cultures could represent either
of these events by a variety of mechanisms.
Several studies have demonstrated the cytotoxic effect of mononuclear cells from patients with arthritis of varying etiology towards homologous fibroblasts in tissue culture (17-22) but cytotoxicity of joint fluid
cells towards autologous peripheral lymphocytes has not to our knowledge been re1i8
ported. A complement dependent cytotoxic
antibody which is specific for thymus dependent cells (T-cells) has been found to
occur in serum from patients with rheumatoid arthritis and systemic lupus erythematosus (13). It is of interest, that in all
four cases in which this decreased incorporation was observed, the presence of cytotoxic factors against normal lymphocytes in
the respective plasma samples was also demonstrated.
Our results do not exclude the possibility that the observed interactions between
the two cell populations are nonimmunologic or due to a contaminating cell type
in one or the other preparations. More
detailed studies directed towards further
purification and identification of specific
cell types will be required before this reservation can be eliminated. A second factor
which should be considered is the doseresponse relationship which exists in mixed
lymphocyte cultures. It is possible that other combinations of the two cell populations
could present a quite different picture.
It would be premature, at this time, to
interpret the data too closely with respect
to the exact nature of the interaction which
is occurring between synovial fluid cells
and peripheral blood lymphocytes and also
with regard to extrapolation of in vitro
events to the in vivo situation. Suffice it to
say, the marked stimulation of lymphocytes
from inflammatory synovial fluids by mitomycin-treated peripheral blood lymphocytes documented here was a surprising
finding and opens the way for a more intensive study of the problem of cell interactions in the connective tissue diseases.
Conditions such as rheumatoid arthritis are
particularly suitable for this type of study
since cells from the fluid bathing the lesions (synovial fluid cells) are available for
study and comparison to peripheral blood
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
LYMPHOCYTE INTERACTIONS
lymphocytes. We feel that several interesting avenues of investigation into the types
of cell to cell interactions which occur in
inflammatory arthritis are indicated and
will require closer examination in future
studies if we are t o understand the local
immune or inflammatory reaction i n rheumatoid arthritis.
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GElGY RHEUMATISM PRIZE
Dr. K. Frank Austen, physician in chief at the Robert B. Brigham Hospital, Boston, and first incumbent of the Theodore B. Bayles Professorship
on the faculty of medicine at Harvard Medical School, received the International Geigy Rheumatism Prize for his studies on the immunobiology and
immunochemistry of complement. T h e Geigy Prize is awarded every 4 years
by the International League Against Rheumatism.
120
Arthritis and Rheumatism, Vol. 17, No. 2 (March-April 1974)
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