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Infection and Immunity, July 1999, p. 3221-3226, Vol. 67, No. 7
Department of Immunology,
Received 17 August 1998/Returned for modification 14 October
1998/Accepted 31 March 1999
In active tuberculosis, T-cell response to Mycobacterium
tuberculosis is known to be reduced. In the course of
Mycobacterium tuberculosis infection in mice, we observed
that T-cell proliferation in response to M. tuberculosis
purified protein derivative (PPD) reached the maximum level on day 7, then declined to the minimal level on day 14, and persisted at a low
level through day 28 postinfection. The frequency of PPD-specific CD4 T
cells in the spleen on day 28 decreased to one-sixth on day 7. To
further investigate the mechanism of this T-cell hyporesponsiveness, we
next analyzed the suppressive activity of spleen macrophages on T-cell
function. The nonspecific proliferative response of naive T cells and
the PPD-specific proliferative response of T cells were suppressed by
day 28 macrophages, but not by day 7 macrophages or naive macrophages. This reduction of proliferative response was restored by addition of
nitric oxide synthesis inhibitor,
NG-monoethyl-L-arginine monoacetate, but not by
monoclonal antibody against interleukin 10 or transforming growth
factor Mycobacterium
tuberculosis is an acid-fast intracellular pathogen that resides
mainly in the macrophages of hosts and that causes a chronic infection.
It is known that M. tuberculosis induces T-cell-mediated
immunity. The primed mycobacterium-reactive T cells, which consist
mainly of CD4 T cells, activate infected macrophages in
granulomatous lesions by Th1-type cytokines (21, 34). The
activated macrophages subsequently produce bactericidal effector
molecules, such as nitric oxide (6, 9, 16). Thus, the
interaction between T cells and macrophages is critical for prevention
of bacterial growth (4, 8, 34). It is also possible that
macrophages in infected sites influence the T-cell response in vivo.
Our group and others have previously reported that induction of
T-cell-mediated immune response was highly influenced by the activated
macrophages (26, 30, 31, 36, 45, 48).
Some patients with tuberculosis showed depressed immune response
(41). Depressed proliferative response and interleukin 2 (IL-2) production of PBMC against purified protein derivative (PPD) of
M. tuberculosis (25, 43) was found in 40 to 60%
of patients with active pulmonary tuberculosis. In experimental murine tuberculosis, it was reported that gamma interferon (IFN- The purpose of this study is to determine the mechanisms of T-cell
hyporesponsiveness during M. tuberculosis infection. We report here that, in murine experimental tuberculosis, T-cell-mediated immune response was suppressed in the chronic stage of infection. The
hyporesponsiveness was associated with a reduced number of mycobacterium-specific CD4 T cells. We also show that the activated macrophages from mice at day 28 postinfection suppressed T-cell response to anti-T-cell receptor (TCR) monoclonal antibodies (MAb) and
PPD by NO.
Mice.
Female C57BL/6 mice bred under specific-pathogen-free
conditions were purchased from Japan SLC (Shizuoka, Japan). Seven- to 9-week-old mice were used for experiments.
Bacteria and infection.
Mice were infected intravenously
with 106 CFU of M. tuberculosis (H37Rv)
harvested at the mid-log phase of growth in Middlebrook 7H9 medium
(Difco Laboratories, Detroit, Mich.) supplemented with 0.05 mg of oleic
acid (Wako Pure Chemical Industries Ltd., Osaka, Japan)/ml, 0.05%
Tween 80 (Difco), 2 mg of dextrose/ml, 0.85 mg of NaCl/ml, and 5 mg of
bovine serum albumin (Sigma Chemical Co., St. Louis, Mo.)/ml and kept frozen.
Preparation of lymphocytes and macrophages.
Spleens were
aseptically removed from mice and teased between two sterile glass
slides. After erythrocytes were lysed by treatment with 0.83% ammonium
chloride in 0.17 mM Tris-HCl (pH 7.6), the spleen cells were washed
twice with Hanks balanced salt solution and suspended in RPMI 1640 medium (Gibco, Grand Island, N.Y.) supplemented with 10% fetal calf
serum (Hyclone, Logan, Utah), 50 µM 2-mercaptoethanol, 20 µM HEPES,
and 0.02% sodium bicarbonate. Nylon wool-passed cells and
plastic-adherent cells were used as T cells and macrophages,
respectively. Lymph node T cells were prepared from the mesenteric
lymph nodes of uninfected mice. In some experiments, CD4 T cells were
enriched from T cells by depletion of CD8 T cells with a supernatant of
anti-CD8 MAb-producing hybridoma (83.12.5) plus complement.
Proliferation assay.
T cells (2 × 105/well) prepared from spleens were cultured in 96-well
tissue culture plates (Coster, Cambridge, Mass.) in triplicate, with or
without 10 µg of PPD of M. tuberculosis (Aoyama strain) (Japan BCG Inc., Tokyo, Japan), for 72 h in the presence of 4 × 105 irradiated splenocytes/well as antigen-presenting
cells (APC). In some experiments, recombinant human IL-2 (rIL-2;
generously provided by Takeda Chemical Industries Ltd., Osaka, Japan)
was added. Whole spleen cells (4 × 105/well) instead
of purified T cells were used in proliferation assays in some
experiments. Cultures were pulsed with 1 µCi of [3H]thymidine for the last 6 h of culture, and
[3H]thymidine uptake was measured by liquid scintillation counting.
Cytokine measurements.
T cells (106/well) from
spleens were cultured in 24-well tissue culture plates in triplicate,
with or without 10 µg of PPD/ml for 72 h in the presence of
2 × 106 irradiated splenocytes/well, and the culture
supernatants were collected and used for cytokine measurements by
standard sandwich enzyme-linked immunosorbent assay (ELISA), as
described in a previous report with minor modification (24).
Briefly, the supernatants were added to the wells of enzyme immunoassay
plates (Greiner, Frickenhausen, Germany), precoated overnight with 2 µg of rat anti-mouse IFN- Limiting-dilution assay.
The frequency of PPD-reactive cells
was determined by limiting-dilution assay as previously described
(5, 29). Briefly, CD4 T cells prepared from spleens were
plated in 96-well tissue culture plates in 12 replicates for each
concentration and cultured for 72 h with or without 10 µg of
PPD/ml in the presence of APC. The cultures were pulsed with 1 µCi of
[3H]thymidine, and [3H]thymidine uptake was
measured by liquid scintillation counting. Wells of culture with PPD
were defined as positive when [3H]thymidine uptake was
higher than the mean ± 3 standard deviations (SD) of Ag-negative
wells containing the same cell concentrations.
Inhibition of T-cell-proliferative response by macrophages.
Splenocytes were analyzed for their Mac-1 expression by
fluorescence-activated cell sorter and plated in 96-well-plates, each containing 3 × 105 Mac1-positive cells as
macrophages. The plates were then incubated for 60 min to allow the
macrophages to adhere to the plates and washed twice with warm
phosphate-buffered saline to remove nonadherent cells. Naive lymph node
T cells (5 × 104/well) were added to the macrophage
monolayer and cultured for 72 h in the presence of 20 µg of
anti-TCR NO assay.
Macrophages prepared by the method mentioned above
were cultured for 72 h with medium only. The production of NO was
estimated by measuring the amount of nitrite in the culture supernatant according to the method of Green et al. (17) with Greiss
reagent (Wako). Absorbance was measured with a microplate reader. The titer was determined by the standard curve generated by the absorbance of serial dilutions of NaNO2.
Statistical analysis.
Student's t test (two
tailed) was used for statistical analysis; a P value of less
than 0.05 was considered significant.
Kinetics of PPD-reactive T-cell response after infection with
M. tuberculosis.
In the first set of experiments, we
examined the M. tuberculosis-specific proliferative response
of spleen T cells in mice during the course of M. tuberculosis infection (Fig. 1). T
cells obtained on day 7 of infection showed a high level of
proliferation in response to PPD in the presence of naive syngeneic
irradiated splenocytes. On day 14 of infection, however, the T-cell
proliferative response to PPD was significantly decreased, and the low
level of T-cell proliferative response persisted through day 28 of
infection. This reduced response cannot be explained by typical T-cell
anergy, since the addition of 20 (experiment 1) and 2 (experiment 2) U of rIL-2 to the cultures failed to restore the proliferative response on day 14 and day 28. Since a high number of mycobacteria was detected
in spleens on both day 7 [(4.3 ± 0.8) × 106
CFU/spleen], and day 28 [(2.8 ± 0.6) × 106
CFU/spleen], the lack of responsiveness on day 28 cannot be explained by differences in bacterial load or amount of Ag. The suppressed proliferative response of spleen T cells to PPD in mice infected with
M. tuberculosis is observed for at least 90 days after
infection (data not shown).
Kinetics of cytokine production by T cells.
To determine if T
cells on day 28 produce suppressive cytokines in an Ag-specific manner,
culture supernatants were assayed for cytokine production by the
standard sandwich ELISA (Table 1).
IFN-
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
T-Cell Hyporesponsiveness Induced by Activated
Macrophages through Nitric Oxide Production in Mice Infected with
Mycobacterium tuberculosis
and
ABSTRACT
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
. These data indicate that the macrophages from mice
chronically infected with M. tuberculosis suppress T-cell
response through production of nitric oxide, suggesting that nitric
oxide-induced elimination mediated by activated macrophages may reduce
the T-cell response and the number of mycobacterium-specific CD4 T
cells in vivo.
INTRODUCTION
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
)
production by CD4 T cells declined after the third or fourth week of
infection (35). Nonspecific response to concanavalin A or
antigen (Ag)-specific proliferative response by splenocytes also
declined after mycobacterial infection (36). Other chronic
disease pathogens, such as Leishmania, Trypanosoma, and Toxoplasma, are also reported to
induce T-cell hyporesponsiveness (23, 28, 39, 46). Nitric
oxide derived from macrophages was reported to be a major cause of
T-cell suppression in murine Salmonella (3, 10,
11) and Trypanosoma (38) infections. In
M. tuberculosis infection in mice, the mechanisms of the
reduction of T-cell responses are not clearly understood. Furthermore,
it remains unknown whether the immune suppression contributes to
exacerbation of the disease.
MATERIALS AND METHODS
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
MAb (R4-6A2) (PharMingen, San Diego,
Calif.) or rat anti-mouse IL-4 MAb (11B11) (PharMingen). After
incubation for 60 min at room temperature, the plates were washed with
phosphate-buffered saline containing 0.05% Tween 20 and incubated at
room temperature for 60 min with 2 µg of biotin-conjugated anti-mouse
IFN- (XMG1.2) (PharMingen) or IL-4 MAb (BVD6-24G2) (PharMingen)/ml.
The plates were then washed, and streptavidin--galactosidase (Gibco)
was added at a dilution of 1/1,000 to each well and incubated for 45 min. After the plates were washed, substrate solution containing 0.2 mM
4-methylumbelliferyl--D-galactopyranoside (Wako) was
added to the wells, which were left for 45 to 60 min while being
protected from direct light. Finally, after the addition of 0.1 M
glycin-NaOH (pH 10.2), absorbance was measured with a fluorescence
microplate reader (MTP-32; Corona Co., Ltd., Ibaragi, Japan). The
values for IFN- and IL-4 were calculated from a standard curve of
recombinant mouse IFN- and IL-4 (PharMingen).
MAb (H57-597)/ml and 20 µg of streptomycin (Meiji)/ml.
In some experiments, spleen T cells from infected mice on day 7 were
used instead of naive lymph node T cells and stimulated with 10 µg of
PPD/ml instead of anti-TCR MAb. The cultures were pulsed with 1 µCi of [3H]thymidine, and [3H]thymidine
uptake was measured by liquid scintillation counting. To neutralize the
cytokines, anti-IL-10 (SXC-1) (PharMingen), and anti-pan transforming
growth factor (TGF-) (R & D Systems, Minneapolis, Minn.) were
added at 20 µg/ml. Ten millimolar
NG-monoethyl-L-arginine monoacetate (L-NMMA)
(Wako) and NG-monoethyl-D-arginine monoacetate
(D-NMMA) (Wako) were used as an inhibitor of NO synthase and its
control, respectively.
RESULTS
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Abstract
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Materials and Methods
Results
Discussion
References
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FIG. 1.
Kinetic analysis of proliferative response to PPD by
spleen T cells from mice infected with M. tuberculosis. The
mice were infected intravenously with 106 CFU of M. tuberculosis H37Rv. On the indicated days, nylon wool-passed
spleen cells (2 × 105) from four to six spleens were
cultured in triplicate with or without 10 µg of PPD/ml for 72 h
in the presence of irradiated syngeneic naive splenocytes as APC and 20 (experiment 1) or 2 (experiment 2) µg of rIL-2/ml was added to some
of the cultures. The cultures were pulsed with 1 µCi of
[3H]thymidine for the last 6 h of culture, and
[3H]thymidine uptake was measured by liquid scintillation
counting. The results are presented as the mean values of triplicate
wells ± SD. *, P < 0.05 versus T cells
cultured with PPD on days 0, 14, and 28. **, P < 0.05 versus T cells cultured with PPD plus IL-2 on days 0, 14, and
28. Exp., experiment.
production in response to PPD was detected from day 7 and
reached a peak on day 14. On day 28 of infection, IFN- production by
spleen T cells was decreased compared to that on day 14, but
significant IFN- production was still detected. IL-4 (Table 1) and
IL-10 (data not shown), which were thought to be suppressive for Th1
response, were not detected in the samples. These data did not support
Th2-type T-cell influence on T-cell proliferation and IFN-
production on day 28. It is notable that there were high levels of
IFN- production on day 14 and day 28, when PPD-specific
proliferative response was low.
TABLE 1.
Kinetics of cytokine production by T cells in mice
infected with M. tuberculosis
Frequency of PPD-reactive CD4 T cells in spleens after infection with M. tuberculosis. We next checked if the frequency of PPD-responsive T cells had decreased in vivo on day 28 of infection. As shown in Fig. 2, we found, by limiting-dilution assay, that the frequency of PPD-reactive CD4 T cells in spleens on day 28 (1/106,415) was lower than that on day 7 (1/16,904). This result suggests that the number of mycobacterium-reactive CD4 T cells in the spleen declines in the course of M. tuberculosis infection from day 7 to day 28.
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Inhibition of T-cell proliferation induced by macrophages from mice
infected with M. tuberculosis.
Since it was reported that
activated macrophages induce T-cell apoptosis (31), we
hypothesized that the activated macrophages on day 28 of infection can
kill or suppress the mycobacterium-specific T cells. To address this
hypothesis, we analyzed the inhibition of T-cell stimulation by the
macrophages from mice infected with M. tuberculosis by using
an in vitro system. In the experiments, naive lymph node T cells were
stimulated by anti-TCR MAb in the presence of spleen macrophages
derived from naive mice or M. tuberculosis-infected mice on
day 7 and day 28. As shown in Fig. 3,
although there was high proliferative response of naive T cells to
anti-TCR MAb in wells to which naive or day 7 macrophages were added,
the T-cell proliferative response significantly decreased in the
presence of day 28 macrophages. Furthermore, we found that PPD-specific
proliferative response of spleen T cells from mice on day 7 postinfection significantly decreased when they were cultured with day
28 macrophages (Fig. 4B).
|
|
). (+), present; (
),
absent.
Inhibition of T-cell proliferation induced by day 28 macrophages
was restored by L-NMMA.
From the data mentioned above, it is
possible that day 28 macrophages actively suppress T-cell response. NO
is a possible suppressor because NO is produced mainly by activated
macrophages in infections with mycobacteria (33, 47) or
other intracellular parasites (37). NO is not only a
bactericidal effector molecule (6, 7, 9, 16) but also a
suppressive factor for Th1 (39, 40, 46). Other candidates
are macrophage-derived suppressive cytokines, such as TGF- and
IL-10. We studied the effects of blocking these substances on
suppression of T-cell response mediated by day 28 macrophages.
MAb-stimulated T-cell proliferation when
T cells were cultured with day 28 macrophages were restored by the
addition of L-NMMA, an NO synthase inhibitor (Fig. 4A). Those of
PPD-specific T cell proliferation when T cells were cultured with day
28 macrophages were partially restored by the addition of L-NMMA (Fig.
4B). In contrast, an isomer of L-NMMA, D-NMMA, which lacks NO
synthesis-blocking activity, showed no effect on the T-cell responses.
Furthermore, macrophage-derived nitrite on day 28 of infection
(4.40 ± 1.19 µM), measured in culture supernatant, was
approximately fourfold higher than that from naive macrophages (1.26 ± 0.44 µM) or that from macrophages on day 7 (1.45 ± 0.29 µM). In contrast, cytokine-neutralizing experiments showed
that neither anti-IL-10 nor TGF- MAb restored the T-cell
proliferative response against anti-TCR MAb (Fig. 4A) or PPD (Fig.
4B). These data strongly suggest that day 28 macrophages suppress the
T-cell proliferation through NO production. The absolute cpm values of proliferative responses shown in Fig. 4B were lower than those in Fig.
4A, since PPD stimulation was weaker than mitogenic stimulation of
anti-TCR MAb. In addition, the Ag (PPD)-presenting capacity of
macrophages was considered to be lower than that of irradiated splenocytes, as shown in Fig. 1.
The effect of L-NMMA on the proliferative response of total spleen cells from mice infected with M. tuberculosis. To demonstrate the significance of NO in an in vivo situation, we analyzed the proliferative response of total spleen cells in response to PPD (Fig. 5). On day 28 of infection, when NO production from macrophages was increased, L-NMMA did not restore the proliferation in response to PPD. This result may be explained by NO-mediated elimination of PPD-reactive T cells on day 28 of infection, as shown in Fig. 2. Interestingly, L-NMMA restored the proliferative response of day 7. This result is probably due to the low levels of NO produced by macrophages, which can suppress T-cell proliferation but are not sufficient to eliminate T cells.
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Failure to transfer the suppressive effect on proliferative response by supernatants obtained from cultures with activated macrophages. To investigate whether the suppressive effect of macrophages on T cells is transferable by culture supernatant, we performed the following experiment. The macrophages from mice infected with M. tuberculosis on day 90 were cultured for 72 h, and the supernatant was collected. The supernatant was transferred to the wells with the whole spleen cells from mice on day 7 of infection and cultured for 72 h with PPD or anti-TCR MAb. Transferred supernatant had no suppressive effect against the proliferative response to the stimuli (data not shown). This result is consistent with NO-mediated suppression, because the half-life of NO is too short for it to transfer by culture supernatant.
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DISCUSSION |
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Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Our study demonstrated that activated macrophages obtained on day 28 from mice infected with M. tuberculosis have strong suppressive activity on T-cell response against mycobacterial Ag and mitogenic anti-TCR MAb. Furthermore, the frequency of mycobacterial Ag-reactive T cells in the spleen on day 28 of M. tuberculosis infection was decreased compared to that at earlier stages of infection. These observations suggest that activated macrophages in mice chronically infected with M. tuberculosis suppress T-cell response, which may subsequently induce apoptosis of mycobacterial Ag-reactive T-cell populations. There are several possible mechanisms to explain the macrophage-mediated suppression of T-cell response, as discussed below.
Our results show that NO is involved in the suppression of T-cell response by the macrophages from mice chronically infected with M. tuberculosis. Macrophages from M. tuberculosis-infected mice on day 28 produced three to four times more NO than macrophages obtained on day 7 of infection and those from naive mice. Blocking of NO synthesis by L-NMMA restored the proliferative response of T cells cultured with the day 28 macrophages. These observations demonstrated that the large amount of NO produced by the suppressor macrophages on day 28 of M. tuberculosis infection suppresses T-cell response in vitro. It has been shown in other systems that NO contributes not only to killing pathogens but also to suppressing T-cell responses, especially Th1-type responses (39, 40, 46), which further supports our data showing NO-mediated T-cell suppression. In the present study, the restoration of proliferative response of total spleen cells by L-NMMA was not observed on day 28 (Fig. 5), when a larger amount of NO was produced by macrophages. This was explained by the reduced number of PPD-reactive T cells on day 28 of infection in vivo. On the other hand, it is surprising that L-NMMA restored PPD-specific proliferation on day 7 (Fig. 5), although NO production by macrophages was at low levels on day 7 of infection. We can explain this discrepancy by the fact that NO-induced T-cell suppression is observed even on day 7 with a small amount of NO, although the number of PPD-specific cells is not reduced.
NO-induced apoptosis may explain both the in vivo decline in the frequency of mycobacterial Ag-reactive CD4 T cells in the spleen observed on day 28 of M. tuberculosis infection and the in vitro macrophage-mediated suppression of T-cell response. There are several reports showing NO as the mediator of apoptosis. NO has been shown to induce apoptosis of human lymphoblastoid cells (32), mouse thymocytes (13), mouse peritoneal macrophages (1), rat lung epithelial cells (19), rat hepatoma cells (27), and rat islet cells (12), possibly through excessive activation of poly(ADP-ribose) polymerase and a consequent depletion of its substrate NAD+ (18). Apoptosis was also induced on peripheral T cells by macrophage colony-stimulating factor-stimulated macrophages in a Fas- and Bcl-2-independent manner when the T cells recognized Ag expressed by the macrophages (31). Therefore, it is highly possible that apoptosis is also induced in T cells by the high level of NO produced by the activated macrophages from mice chronically infected with M. tuberculosis both in vivo and in vitro. However, it is still debatable whether in vitro suppression of naive T-cell response by macrophages from chronically infected mice, as shown in Fig. 3 to 5, is accompanied by apoptosis. We stained T cells cultured with macrophages from naive or infected mice on day 7 or day 28 with an in situ nicking and end-labeling technique and used flow cytometry to detect apoptotic cells. However, the T-cell preparation contained a large amount of dead cells and cell debris even in the control culture with naive macrophages, which made the analysis difficult (data not shown). More sensitive methods to detect apoptotic cells or an improved culture system that induces less non-specific cell death would be required to detect differences of apoptosis in vitro.
Another possible mechanism of macrophage-mediated suppression of T-cell response is the lack of costimulation of T cells. It is known that T-cell activation in the absence of costimulation by the B-7-CD28 system induces T-cell anergy (22). Saha et al. reported that B-7 expression on BALB/c macrophages was downregulated after in vitro M. tuberculosis infection (36), which supports T-cell hyporesponsiveness induced by the lack of costimulation after M. tuberculosis infection. In our M. tuberculosis in vivo experiments, there is a possibility that the lack of costimulation may induce T-cell anergy on day 28 of infection. Further analysis of B-7 expression on macrophages and CD28 expression on T cells is needed. However, it is suggested that anergy induced by the lack of costimulation may not be the cause of T-cell suppression, because addition of IL-2 failed to restore the proliferation of T cells from mice on day 28 postinfection (Fig. 1). Previously, it was also reported that the addition of IL-2 did not restore the suppression of anti-sheep erythrocyte plaque-forming cell response of spleen cells after infection with Salmonella typhimurium in mice (2).
Macrophages are also known to produce cytokines that suppress T-cell
responses. It was reported, in patients with active tuberculosis, that
blood monocytes release molecules suppressive for T-cell response, such
as soluble IL-2 receptor (44) and TGF- (42). The monocyte-derived TGF- was shown to suppress proliferation and
IFN- production by peripheral blood mononuclear cells in response to
PPD (20). It was also reported that IL-10, another potent
cytokine suppressive for Th1 response (14), was produced by
macrophages infected with mycobacteria (15). However,
neutralization of TGF- and IL-10 did not restore the T-cell
hyporesponsiveness induced by the macrophages from mice chronically
infected with M. tuberculosis. Prostaglandins are another
candidate for molecules which suppress T-cell response. At present,
there is no direct data on the role of prostaglandins in T-cell
suppression in our system, because we did not use indometacin to block
the synthesis of prostaglandins in experiments such as those shown in
Fig. 1, 4, and 5. However, we do not consider that prostaglandins are important in the suppression of T-cell response, because culture supernatant obtained from the culture with the activated macrophages from mice on day 90 of infection did not suppress the proliferative response of spleen cells from mice on day 7 of infection. These observations do not support T-cell suppression caused by cytokines produced by macrophages.
Although our results clearly demonstrate low T-cell proliferative
response against PPD in the spleens of chronically M. tuberculosis-infected mice and T-cell suppression by macrophages,
it is debatable whether the immunosuppression contributes to
persistence of the infection or exacerbation of the disease. The number
of bacteria in the spleens of M. tuberculosis-infected mice
showed no increase on day 28 of infection, although the immune response
of the spleen was significantly suppressed. Evidence for the
bactericidal effect of NO in mycobacterial infection in mice has been
reported by Chan et al. (6). Although NO is necessary for
host defense, the significance of NO-mediated T-cell suppression in
host defense is unknown. It has been reported that protective cells
were induced and maintained even if NO-mediated suppression was
demonstrated in vaccination with some Salmonella strains
(11). In the present study, it is noteworthy that the
IFN- production of spleen T cells was clearly demonstrated in
response to PPD when the proliferative response to PPD was low.
Therefore, protective immunity against mycobacteria could be maintained
through IFN- production, even when the proliferative response of T
cells to mycobacterial Ag is decreased. These IFN--producing T cells
are considered to be protective and independent of proliferation, and
to have the capacity to avoid apoptosis. To confirm this hypothesis, we
need further analysis of T-cell function in the chronic phase of
M. tuberculosis infection by such tests as ELISPOT assay of
IFN- production.
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ACKNOWLEDGMENT |
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This work was supported in part by Japan-U.S. Cooperative Medical Science.
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FOOTNOTES |
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* Corresponding author. Present address: Department of General Medicine, Kyushu University Hospital, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812, Japan. Phone: 81 92 642 5909. Fax: 81 92 642 5916. E-mail: snabe{at}genmedpr.med.kyushu-u.ac.jp.
Present address: Department of Virology, School of Medicine, Kyushu
University, Fukuoka 812, Japan.
Editor: S. H. E. Kaufmann
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REFERENCES |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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