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Studies of immune functions of patients with systemic lupus erythematosus. i. dysfunction of suppressor t-cell activity related to impaired generation of rather than response to suppressor cells

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657
STUDIES OF IMMUNE FUNCTIONS OF
PATIENTS WITH SYSTEMIC LUPUS
ERYTHEMATOSUS
I. DYSFUNCTION O F SUPPRESSOR T-CELL ACTIVITY
RELATED TO IMPAIRED GENERATION OF, RATHER
THAN RESPONSE TO, SUPPRESSOR CELLS
TSUYOSHI SAKANE, ALFRED D. STEINBERG, and IRA GREEN
T cell suppressor function in patients with systemic
lupus erythematosus (SLE) was evaluated by studying the
ability of concanavalin A- (Con A) activated T cells to
suppress proliferative responses by responder cells autologous with the Con A-activated T cells. Impaired suppressor T-cell activity in patients with SLE was observed with
regard to three effector functions: 1) the allogeneic response of T cells, 2) the Con A response of T cells, and 3)
the B cell response to pokeweed mitogen (PWM). No
defect was found with regard to suppression of the T-cell
response to phytohemagglutinin (PHA).
Mixing experiments between SLE and normal
cells were carried out to further define the nature of the
SLE suppressor defect. When responder cells from SLE
patients were cultured with Con A-activated T cells from
normal controls, the normal suppressor T cells caused
suppression of SLE responder cells. In contrast, Con AFrom the Laboratory of Immunology, National Institute of
Allergy and Infectious Diseases, and The Arthritis and Rheumatism
Branch. National Institute of Arthritis, Metabolism and Digestive
Diseases, National Institutes of Health, Bethesda, Maryland 20014.
Supported i n part by Cancer Research Institute Incorporated, New York, New York.
Tsuyoshi Sakane, M.D.: Visiting Fellow, Laboratory of Immunology, NIAID: Alfred D. Steinberg, M.D.: Senior Investigator,
Arthritis and Rheumatism Branch, NIAMDD; Ira Green, M.D.:
Senior Investigator, Laboratory of Immunology, NIAID.
Address reprint requests to Tsuyoshi Sakane, M.D., Ph.D.,
Department of Health, Education, and Welfare, Public Health Service, National Institutes of Health, Building 10, Room 1 IN-314, Bethesda, Maryland 20014.
Submitted for publication December 16, 1977; accepted in
revised form March 13, 1978.
Arthritis and Rheumatism, Vol. 21, No. 6 (July-August 1978)
activated T cells from SLE patients were incapable of
exerting suppressor effects on normal responder cells.
These observations indicate that the impaired suppressor
activity in SLE patients resides in the generation of suppressor T cells, rather than in the response to suppressor T
cell signals.
Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown cause. Viral, genetic,
hormonal, and immunologic factors have been implicated in the pathogenesis of the illness (1-17). The common pathway to disease appears to operate through the
immune system. Immune abnormalities found in patients with active SLE include impaired proliferation of
lymphoid cells in response to mitogens, antigens, and
allogeneic cells (7), hyporesponsiveness to intradermal
injection of antigen (8), and impaired primary antibody
responses (9). Patients with active SLE also produce a
variety of antibodies reactive with self antigens including those on T lymphocytes (10-12). Their excessive Bcell activity includes hypergammaglobulinemia ( 13) and
circulating plaque forming cells producing antibody to
nucleic acids (14) and chemical haptens ( 1 5 ) . The reason
for the development of autoreactive antibodies as well as
the B cell hyperactivity is uncertain, but could be explained by a relative loss or dysfunction of suppressor
cells which may regulate the responses of both T and B
cells. Such cells have been well studied in animals (19)
and have also been described in several human studies
( 16-25).
S A K A N E ET A L
658
T o investigate the suppressor cell function in
SLE patients, we have taken advantage of the observations of Shou et al. (23) that human peripheral blood
lymphocytes can be induced by concanavalin A (Con A)
to manifest suppressor functions for proliferative responses in vitro. The precursors of such Con A-activated suppressor cells were shown in our previous studies to be T cells (25). Furthermore, blastogenic response
to Con A was not necessary for the generation of suppressor cells (25). On the basis of these studies it became
possible to assess T-cell suppressor function by studying
the ability of purified T cells from SLE patients activated by Con A to suppress both T cell- and B cellproliferative responses. In the present studies, we have
utilized this system to investigate the role of suppressor
T cells in SLE. In particular, we have asked two questions: 1 ) Can patients with SLE generate normal suppressor function? 2) Can SLE cells respond to normal
suppressor signals? We found that SLE patients can
respond normally to suppressor signals, but that they
have some defects in generating suppressor cell activity,
resulting in a variable impairment in suppressor function.
MATERIALS AND METHODS
Patient Selection. Ten patients with well documented
SLE, followed by the Arthritis and Rheumatism Branch, National Institutes of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health, were selected for the
present studies. All patients satisfied the diagnostic criteria of
the American Rheumatism Association for SLE. Their mean
age was 31 years and 9 patients were female. Patients receiving
immunosuppressive drugs or high doses of corticosteroids
(> 10 mg/day prednisone) were excluded. Disease activity was
evaluated based upon clinical status and graded as active,
mildly active, or inactive. Age and sex matched controls consisted of 7 healthy adult subjects whose blood was provided
from the Blood Bank Department, Clinical Center, National
Institutes of Health,
Separation of Purified T and B Cells. Procedures for
separation of purified T and B cells were given in detail previously (25). Only a brief outline is presented below. Mononuclear cells were isolated from heparinized peripheral blood
over a Ficoll-Hypaque gradient. These cells were rosetted with
sheep erythrocytes (SRBC) and the rosetted lymphocytes were
separated on another Ficoll-Hypaque gradient from rosetted
lymphocytes. The SRBC in each fraction were lysed with
ammonium chloride buffer. The unrosetted cell fraction in
complete culture medium, RPMI 1640 (Grand Island Biological Co., Grand Island, New York), and 10% fetal bovine
serum (Microbiological Associates, Bethesda, Maryland) was
plated o n petri dishes at 37°C for 2 hours, and both adherent
and non-adherent cells were then collected separately. T h e cells
which formed rosettes with SRBC will be referred to as T
lymphocytes, the non-adherent cells which did not form rosettes
will be designated, for convenience, as B lymphocytes, and the
adherent cells as monocytes. The T lymphocyte preparation
contained more than 95% rosetted lymphocytes, whereas in the
B cell fraction less than 3% of the cells formed rosettes with
SRBC. Adherent monocyte preparations consisted of 95%
cells that were identified as monocytes in Giemsa stained
smears.
Overall Experimental Design. Suppressor T cells were
generated by Con A activation in a first culture. These cells
were added t o responder cells in a second assay culture system
that were stimulated with either mitogens or allogeneic cells.
In the majority of studies, the suppressor cells from the first
culture and the responder cells in the second culture were from
the same individual. In certain experiments, suppressor cells
from one individual were mixed with responder cells from
another individual.
Generation of Suppressor T Cells by Activation with
Con A (First Culture). Five X I @ T lymphocytes were incubated in 5 ml complete culture medium alone (non-activated
control T cells) or with 50 pg Con A (Con A-activated T cells)
(Pharmacia Fine Chemicals, Piscataway, New Jersey). To
both non-activated control and Con A-activated cultures, 0.3
X 108 mitomycin C (Sigma Chemical Co., St. Louis, Missouri)-treated monocytes were added. Sixty hours later, the
cells were harvested, washed four times, treated with mitomycin, and then tested for their suppressor activity in the second
assay culture system.
Assay for Suppressor Activity of Con A-Activated T
Cells in a Second Culture System. Responder cells were obtained 3 days later from either a new bleeding of the same
individual who originally provided the suppressor cells, or
from another individual. Stimulatory rnitogens or allogeneic
stimulating cells were added to 1 X 106 responder cells in
microtiter plates, and subsequently 1 X lo6 mitomycin-treated
Con A-activated or nonactivated control T cells from the first
culture were added. In addition, to fully develop the suppressor activity by Con A-activated cells, mitomycin-treated
monocytes (5,000 per culture) were also added to the second
culture system (26). Where T lymphocytes were used as responding cells, they were stimulated by either phytohemagglutinin (PHA; Wellcome, Beckenham, England) (0. I pg/ml),
Con A (2 pg/ml) or 1 X 10' mitomycin-treated allogeneic
stimulating cells. Here, PHA and Con A were used at the
suboptimal concentration in the second culture system for
determination of suppressor effects on these mitogenic responses. When B lymphocytes were used as responder cells,
pokeweed mitogen (PWM) (Grand Island Biological Company) (2 pg/ml) was the stimulant in the culture. Since the B
cell response to PWM is clearly T-cell dependent, freshly prepared autologous T cells which had been treated with mitomycin were added (5 X IO'/culture). T-cell cultures which were
stimulated by mitogens were harvested on day 3, and T-cell
cultures being stimulated by allogeneic cells, as well as B cell
cultures stimulated with PWM, on day 5 . In both cases, DNA
synthesis in the second culture period was assayed by adding I
pCi of 'H-thymidine (5 Ci/mM; Amersham/Searle Corporation, Arlington Heights, Illinois) to the culture for the final 20
hours in the second culture period. Complete details of these
methods are given in reference 25.
GENERATION OF SUPPRESSOR T CELLS IN SLE
The degree of suppression was calculated with the
following formula:
Mean cpm of
stimulated
cultures con-
Suppression= 1 -
Mean cpm of
unstimulated
cultures containing Con - taining Con
A-activated T
A-activated T
cells
cells
Mean cpm of
Mean cpm of
stimulated
unstimulated
cultures concultures containing non- - taining nonactivated T cells activated T cell;
RESULTS
Suppressor Activity of Con A-Activated T Cells
for the Allogeneic Response by T Lymphocytes. In these
first series of experiments the suppressor cells and the
responder cells were obtained from the same individual.
T lymphocytes from patients with SLE, which had been
pretreated with Con A in a first culture system, were
added to test cultures containing freshly prepared responder T cells and allogeneic mitomycin-treated stimulator cells. In most experiments, the activities of Con Aactivated T cells from a normal donor were assayed
simultaneously with autologous responder T cells.
Con A-activated normal T cells consistently
caused significant suppression of allogeneic stimulation
in the second test culture system. The shadowed area in
Figure 1 shows the mean 9% suppression f 2 standard
deviations produced by Con A-treated normal T cells
from seven individuals (mean, 63.4% suppression; SD
14.8). In contrast to the results with normal lymphocytes, the T lymphocytes from only 3 of 10 patients with
SLE developed normal suppressor activity (Figure 1 ).
The failure of Con A induced proliferation in lymphocytes from patients with SLE cannot be the explanation
for the failure of suppressor cell development because
first, there was no difference in the first culture of the
proliferative response to Con A of the lymphocytes of 7
SLE patients as compared to the proliferative response
of 5 normal individuals (data not shown). Second, our
previous observations demonstrated that blast transformation of T cells by Con A is not necessary for induction of Con A-activated suppressor cells (25).
Suppressor Activity of Con A-Activated T Cells
for the Mitogenic Responses by T Lymphocytes. Con Aactivated T cells were prepared from the patients in the
usual way. These were added to assay cultures of autologous T lymphocytes which were then stimulated with
either PHA or Con A.
659
The mean suppression of the PHA response by
Con A-activated normal T cells is shown in the left part
of Figure 2 by the shadowed area. Except for one patient, T cells from all patients with SLE normally suppressed the PHA response.
In the center part of Figure 2 is shown the suppression of the Con A response. The study of 10 patients
with SLE demonstrated normal suppression in only 3
patients. The cells from the other 7 patients showed a
defect in their lymphocyte suppressor abilities.
Suppressor Activity of Con A-Activated T Cells
for the B Lymphocyte Response to PWM. We also studied the capacity of Con A-activated T cells obtained
from SLE patients to suppress the B-cell proliferative
response to PWM. Con A-activated cells were introduced into assay cultures of B cells obtained from the
CONTROL
PATIENT
1
2
3
4
5
6
7
8
9
10
CPM x 10-3
13.9
IA
A
6.4
13.2
123
12.0
12.1
10.4
16.7
12.5
19.2
MA
A
A
A
A
MA
A
A
0 2 0 4 0 8 0 8 0 1 0 0
PERCENT SUPPRESSION
Figure 1. Percent suppression of the allogeneic response of T cells from
patients with SLE induced by Con A-activated autologous T cells.
Responder T cells from patients with SLE were mixed with autologous T
cells which had been precultured with or without Con A and were then
stimulated for 5 days with allogeneic lymphocytes. The proliferative
response was measured by rhymidine incorporation during the last 20
hours of culture, and percent suppression was calculated as described in
Materials and Methods. The percent suppression is shown for 10 individual SLEpatients. The shaded area shows the mean f 2 S D of suppression
induced by Con A activated T cells from 7 normal individuals studied
at the same time. To show the actual data from which the percent
suppression was calculated, the control cpm-the amount of fhymidine
incorporation by responder SLE T cells to which were added autologous
T cells precultured without Con A and stimulated with allogeneic cellsare shown. The corresponding control response (cpm X lo-') of cells
from 7 normal individuals was 17.9 & 4.7 (mean f standard error of the
mean). I A = inactive SLE; MA = mildly active SLE; and A = active
SLE.
SAKANE ET AL
660
SUPPRESSION OF PWM REs#)NsE
0FBcELI.s
SUPPRESSION OF MKOGENC RESPONSES OF T CELLS
PATIENT
CONTROL
CPM x 103
PHA
14.2
28.1
42.1
16.4
112
20.2
8.4
25.4
14.4
32.4
1 L A
2
A
3
MA
4
A
5
A
6
A
7
A
8 M A
9
10
A
A
CONTROL
CPM x 103
4.6
6.5
P.4
4.5
2.7
10.a
3.4
8.5
11.0
13.5 -20
80100
PERCENT SUPPRESSION
PWM
CanA
o m 4 0 ~ ~ i o oo
m a e o
~
0
20
40
CONTROL
cPMx10-5
10.6
14.0
14.7
13.4
7.0
26.1
8.9
6.7
9.6
80 I 28.7
80 100
Figure 2. Percent suppression of ihe mitogenic responses of S L E lymphocytes induced by Con A-activated autologous T cells. Responder T
cells from ihe SLE paiienis were culiured with autologous T cells preculiured with or without Con A , and were then stimulated with either
PHA or Con A. When suppressor aciivity for ihe PWM response was assayed (righi panel), B cells were used as responding cells. The
percent suppression of each mitogenic response is shown for 10 individual SLEpatients. The paiient numbers compare 10 those in Figure 1.
The shaded area shows ihe mean f 2 standard deviations of suppression induced by Con A-aciivaied T lymphocytes from 7 normal
individuals studied ai the same time. The conirol cpm of responder S L E lymphocyies are also shown as in Figure 1. The corresponding
conirol response (cpm -lo-') by cellsfrom 7 normal individuals were PHA, 30.3 f 3.4; Con A 11.8 f I .6; and P W M 21. I f 3.4 (mean f
siandard error of the mean). IA = inactive SLE; M A = mildly arrive SLE; and A = aciive SLE.
same patient and stimulated with PWM. As shown in
the right part of Figure 2, the T lymphocytes from 8 of
10 patients with SLE failed to manifest a normal degree
of suppressor activity.
Cell Mixing Experiments. In the above experiments, the suppressor cells and the responder cells were
from the same individual. These results in general
showed that, after Con A-activation, the S L E T cells did
not suppress as well as did T cells from normal subjects.
I n an attempt to determine whether or not the impaired
suppressive ability of SLE T cells was in the generation
of suppressor T cells or in the capacity to respond to
negative regulatory signals produced by suppressor T
cells, a series of reciprocal cell mixing experiments between normal and SLE lymphocytes was performed.
First, we show the results of a single experiment
Table 1. Effecis of Normal Con A-Activated T Cells on Miiogenic Responses by S L E Responder T Lymphocytes*
~
Thymidine Incorporation
N o Stimulation
Source of T Cells
SLE
Nonactivated cells
Con A-activated cells
Normal$
Nonactivated cells
Con A-activated cells
Stimulated with PHA
Stimulated with Con A
cpmt
cpmt
% Suppression
cpmt
% Suppression
220 f 30
492 f 42
15,642 f 1,023
11,857 f 440
26.3
4,686 f 235
5,047 f 163
-2.0
420 f 95
446f 5
16,577 f 732
8 , 2 1 9 f 274
51.9
5,330 f 299
1,987 f 131
68.6
* I X I@ responder cells (second culture) from an SLE patient (No. 4 Figure 1) were mixed with T cells whose suppressor activity is being
tested (first column). These T cells were obtained either from this same patient or from a normal individual.
t Data are expressed as mean cpm of triplicate cultures with the standard error of the mean.
$ T h e Con A-activated T cells from the normal individual studied here, when used to suppress the proliferation of his or her own responder
cells in the secoad culture system, gave 41.1% suppression with PHA and 74.0% with Con A.
GENERATION OF SUPPRESSOR T CELLS IN SLE
66 1
Table 2. Effects of Normal Con A-Activated T Cells on Allogeneic Response by SLE
Responder T Lymphocytes*
Thymidine Incorporation
Stimulated with
Autologous Cells
Source of T Cells
SLE
Nonactivated cells
Con A-activated cells
Normal$
Nonactivated cells
Con A-activated cells
Stimulated with
Allogeneic Cells
cpm t
cpmt
% Suppression
1,246 f 63
1,639 f 203
13,542 f 287
13,590f 504
2.8
3,086 & 139
2,897 f 121
12,924 f 710
7,079 f 148
57.5
* I X 106 responder cells (second culture) from an SLE patient (No. 4 Figure I ) were mixed with T
cells whose suppressor activity is being tested (first column). These T cells were obtained either from
this same patient or from a normal individual.
t Data are expressed as mean cpm of triplicate cultures with the standard error of the mean.
$ The Con A-activated T cells from the normal individual studied here, when used t o suppress the
proliferation of his or her responder cells in the second culture system, gave 52.2% suppression with
allogeneic response.
of this type (Tables 1-3) in which the cells of patient
number 4 (see Figure 1 ) with active disease were used as
responder cells or suppressor cells, and the cells of a
normal individual were used only as suppressor cells. T
cells from this SLE patient, preincubated with Con A,
poorly suppressed the response to PHA and Con A
(Table 1). In contrast, normal T cells precultured with
Con A suppressed these responses of SLE responder T
cells. Similarly, in the allogeneic response of SLE T cells,
normal T cells precultured with Con A induced suppression, but SLE T cells did not (Table 2). Normal, but not
SLE, T cells precultured with Con A also suppressed the
proliferative response of SLE B cells to PWM (Table 3).
Table 4 shows a summary of the results of seven
cell mixing experiments between normal and SLE lymphocytes, similar to the experiment shown in Tables 1-3.
In the combination shown on line 3, Con A-activated T
cells from normal cells suppressed the SLE responder
Table 3. Efects of Normal Con A-Activated T Cells on PMN Response by SLE Responder B
Lymphocytes*
Thymidine Incorporation
No Stimulation
Source ofT Cells
SLE
Nonactivated cells
Con A-activated cells
Normal$
Nonactivated cells
Con A-activated cells
cpm t
Stimulated with PWM
cpmt
% Suppression
439 f 16
689 f 90
13,819 f I,O8I
13,224 f 1,258
6.3
1,051 f 99
541 f 1 5
11,441 f 690
7,293 f 710
35.0
* I X 106 responder cells (second culture) from an SLE patient (No. 4 in Figure 1) were mixed with
T cells whose suppressor activity is being tested (first column). These T cells were obtained either
from this same patient or from a normal individual.
t Data are expressed as mean cpm of triplicate cultures with the standard error of the mean.
$ The Con A-activated T cells from the normal individual studied here, when used to suppress the
proliferation of his or her own responder cells in the second culture system, gave 28.1% suppression
with PWM.
SAKANE ET AL
662
Table 4. Summary of Cell Mixing Experiments Between Normal and SLE Lymphocytes
Line
Con A-Activated Responder
No. of
T Cells from
Cells in
Combinations
First Culture Second Culture
Studied
2
Normal
SLE
Normal
SLE
3
4
Normal
SLE
S LE
1
Normal
Percent Suppression*
PHAt
Con A t
Allogeneic Cellst
PWMS
7
10
51.5 f 9.3
60.3 f 6.1
10.6f 1.4
48.9f 7.65
63.4f 5.6
25.1 f 6.8
39.1 f 3.4
9.3 f 6.6
7
6
61.1 f 8.4
46.5 f 6.1
66.9 f 5.6
55.4f 10.1
65.9 f 1.3
22.8 f 8.0
41.4 f 9.1
6.3 f 8.0
* Percent suppression represents the mean f the standard error for designated combination.
t T cells were used as responder cells in the second assay culture.
B cells were used as responder cells in the second assay culture.
indicates significant decrease in suppression by Student's f test (P < 0.05) when compared to suppression observed in normal-normal
combination.
(-)
cells. The degree of suppression was as great as in the
normal cell combination (line l), where both suppressor
and responder cells came from the same normal individuals. Where the SLE cells were used as suppressor cells
and the normal cells as responder cells (line 4), the
suppression generated by Con A-activated SLE T cells
was significantly impaired in the allogeneic and PWM
responses of normal responder cells as compared to the
suppression in the normal-normal combination (line 1 ).
Thus, the failure of lymphocytes from SLE patients to
manifest normal suppressor activity was found to be due
to a defect in production of suppressor signals rather
than a failure in the response to such suppressor signals.
In the above mixing experiments, no stimulation
or only minimal stimulation was observed when either
Con A-activated or nonactivated T cells were cocultured
with allogeneic responder cells in the absence of exogenous stimuli. This is probably explained by the observation of many investigators (27,28) that non-T cells are
the dominant stimulating components in the human
mixed lymphocyte reaction. It should be noted that the
added cells which were being used to assess suppressor
function were highly purified T cells. Also the incubation time of 3 days used in the PHA and Con A
stimulation experiments is too short to generate an optimal mixed lymphocyte reaction.
DISCUSSION
Recently, there has been a proliferation of interest in defining the role of suppressor T cells in normal
and abnormal immune regulation (19). The immune
system requires checks and balances to prevent an excessive reaction by lymphocytes to exogenous antigens. It is
probable that similar immunoregulatory influences play
a crucial role in maintaining tolerance to self antigens.
Thus, the concept that autoimmunity in SLE is related
to the loss of suppressor T cells may provide an explanation for the development of antibodies reactive to
self antigens in the disease process.
The results obtained in the present studies provide some evidence that suppressor T-cell activity in
patients with SLE is in fact impaired for both T- and Bcell functions. In these studies, we utilize a direct in vitro
test system of suppressor T-cell function: the ability of
Con A-activated T cells from SLE patients to suppress
proliferative responses by T and B cells autologous with
Con A-activated T cells. In our total experience, Con Aactivated lymphocytes from all 20 normal subjects studied (the present studies and reference 25) generated suppressor T cells for mitogenic responses of both T and B
cells. Lymphocytes from 18 of these 20 normal controls
were also able to generate suppressor T-cell activity for
the T-cell response to allogeneic cells. In contrast, T
lymphocytes from most SLE patients failed to manifest
such suppressor T-cell activity for the T-cell response to
allogeneic cells, the T-cell response to Con A, and the Bcell response to PWM. No significant impairment in
generation of suppressor T cells was observed for the
PHA response of T cells. The defect in manifestation of
suppressor activity was most profound in the T-cell
response to allogeneic cells and the B-cell response to
PWM. Individual SLE patients varied in their ability to
generate suppressor cells. In some cases cells from a
particular patient failed to suppress in one assay system,
but normally suppressed a different assay system.
Whether this observation is due to different subpopulations of suppressor cells or different sensitivities
of the assays remains to be determined.
In the present studies it was not possible to per-
GENERATION OF SUPPRESSOR T CELLS IN SLE
form dose-response or kinetic studies of the Con A
induction of suppressor cells in the first culture. Therefore it is possible that under different conditions, greater
or less impairment in SLE T-cell function might be
observed.
To determine whether the defect in the suppression phenomena observed in the lymphocytes of patients
with SLE represents a defect in the development of
suppressor cells or a defect in ability of their cells to
respond to suppressor cell signals, cell mixing experiments were performed. Responder cells from either normal individuals or patients with SLE could be suppressed equally by normal T cells activated by Con A.
Thus, the suppressor defect in SLE was not a result of
failure to respond to suppressor signals. However, when
SLE cells were used as the source of the suppressor
signals, there was impaired suppression. That is, compared to suppression by lymphocytes from normal controls, the Con A-activated T cells from SLE patients
produced significantly less suppression of responder
cells from either normal controls or SLE patients. These
cell mixing studies, therefore, suggest that lymphocytes
from patients with SLE fail to generate adequate suppressor T-cell function but still retain the capacity t o
respond to normal suppressor T cells.
The present findings are consistent with previous
observations of impaired suppressor cell function in patients with SLE (16-18). We have extended those observations to suppressor cells, in that T cells from a single
patient were simultaneously studied with regard to their
ability to suppress several in vitro T- and B-cell functions. I n addition, the present studies demonstrate that
the defect of SLE lymphocytes is in the generation of
suppressor cells and not in the response to suppressor
cells. A similar observations has also been made in
NZB/W mice that develop a lupus-like illness associated
with a loss of suppressor T-cell function (29-31).
The reason that most SLE patients appear to lose
suppressor function for some but not other immune
responses remains to be determined. It is also not clear
why suppressor function is frequently lost in these patients. A possible mechanism is the presence of anti-Tcell antibodies that would be capable of inactivating
suppressor cells or their precursors. Several investigators have suggested the existence of antilymphocytic
antibodies in patients with SLE which 1) correlate with
disease activity (32,33) and 2) may be specific for T cells
(1 1,12). Such anti-T-cell antibodies from SLE patients
have recently been shown to eliminate a subpopulation
of T cells (34). Thus, an anti-T-cell antibody could be
responsible for accelerated loss of suppressor cells or
663
their precursors; such a mechanism has been observed in
NZB/W mice (35). Indeed, in preliminary experiments
we have observed that the incubation of normal T lymphocytes with selected SLE sera during the first culture
period with Con A resulted in a failure of these normal
lymphocytes to develop suppressor activity.
Another possible mechanism for suppressor T
cell loss is the binding of immune complexes to Fc
receptor bearing suppressor T cells, thereby causing
them to be dysfunctional or eliminated from the circulation in vivo. Alternatively, an excess of helper T-cell
function, which may be responsible for the abnormal
antibody formation observed in patients with SLE,
might counteract the effects of a normal number of
suppressor T cells. Physical separation of helper and
suppressor T cells will be necessary to resolve these
questions.
ACKNOWLEDGMENTS
The authors are grateful to Dr. Paul V. Holland, Dr.
Richard Davey, and Ms Jane E. Kendall, the Blood Bank
Department, Clinical Center, National Institutes of Health, for
their help and cooperation in supplying the blood from normal
humans used in these studies. W e also wish t o thank Dr. David
W. Alling, the Office of the Scientific Director, National Institute of Allergy and Infectious Diseases, National Institutes of
Health, for help in performing some of the statistical analysis.
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impaired, patients, dysfunction, suppressor, generation, immune, systemic, erythematosus, cells, lupus, response, rather, activity, related, function, studies
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