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Infection and Immunity, December 2000, p. 6954-6961, Vol. 68, No. 12
Laboratory of Cellular Physiology and
Immunology, The Rockefeller University, New York, New York
10021,1 and Department of
Immunology, University of Cape Town, Cape Town, South
Africa2
Received 4 May 2000/Returned for modification 26 June 2000/Accepted 6 September 2000
In experimental mycobacterial infection, tumor necrosis factor
alpha (TNF- The importance of tumor necrosis
factor alpha (TNF- In addition to its essential protective effects in the generation of
immunity against pathogens, TNF- To directly demonstrate the detrimental effects of excess TNF- Mice.
Eight- to 10-week-old C57BL/6 mice (controls),
homozygous TNF- Recombinant BCG.
Cells of recombinant M. bovis
BCG strain Montreal secreting murine TNF- Intravenous infection of mice.
To monitor the course of
infection of mice, two different inocula were used: a high dose of
about 107 organisms or the lower dose of about
105 organisms. Recombinant bacilli in a volume of 200 µl
were injected into the tail vein of mice. The initial infecting load
was assayed by plating liver, spleen, and lung homogenates 6 h
after infection. Thereafter, groups of infected animals were sacrificed
at different time points as indicated. Because mice infected with
higher inocula were likely to die quickly, experiments with high
inocula were terminated earlier (40 days). Mice were first anesthetized
with a solution containing 44 mg of ketamine per kg of body weight (Aveco Co., Inc., Fort Dodge, Iowa) and 5 mg of xylazine per kg of body
weight (Rompum; Mobay Corp., Shawnee, Kans.). Blood was collected by
cardiac puncture, and serum was prepared and kept frozen at
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Immunopathologic Effects of Tumor Necrosis Factor
Alpha in Murine Mycobacterial Infection Are Dose Dependent
ABSTRACT
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
) is required for control of bacillary growth and the
protective granulomatous response, but may cause immunopathology. To
directly examine the positive and detrimental effects of this cytokine,
a murine model was used in which different amounts of TNF- were
delivered to the site of infection. Mice with a disruption in the
TNF- gene (TNF-KO) or wild-type mice were infected with low or high
doses of recombinant Mycobacterium bovis BCG that secreted
murine TNF- (BCG-TNF). Infection of TNF-KO mice with BCG containing
the vector (BCG-vector) at a low dose led to increased bacillary load
in all organs and an extensive granulomatous response in the lungs and
spleen. The mice succumbed to the infection by ~40 days. However,
when TNF-KO mice were infected with low doses of BCG-TNF, bacillary
growth was controlled, granulomas were small and well differentiated,
the spleen was not enlarged, and the mice survived. Infection with high
inocula of BCG-TNF resulted in bacterial clearance, but was accompanied
by severe inflammation in the lungs and spleen and earlier death
compared to the results from the mice infected with high inocula of
BCG-vector. Wild-type mice controlled infection with either recombinant
strain, but showed decreased survival following high-dose BCG-TNF
infection. The effects of TNF- required signaling through an intact
receptor, since the differential effects were not observed when TNF-
receptor-deficient mice were infected. The results suggest that the
relative amount of TNF- at the site of infection determines whether
the cytokine is protective or destructive.
INTRODUCTION
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
) in the host defense against mycobacterial
infection has been appreciated for some time. In experimental animal
models, TNF- production has been shown to be necessary for the
formation and maintenance of the granulomas which seal off foci of
infection and thus limit dissemination of the bacteria. When mice
infected with Mycobacterium tuberculosis received daily
injections of recombinant murine TNF-, a significant reduction in
the number of viable bacteria in the lungs and spleens was observed
(8). Similarly, treatment of Mycobacterium bovis
bacillus Calmette-Guérin (BCG)-infected mice with TNF and
TNF-mimetic peptide (consisting of amino acids 70 to 80 of TNF)
resulted in increased organization of the granulomas and a decrease in
the bacterial load (25). Conversely, when TNF- was
neutralized by treatment with anti-TNF- monoclonal antibody,
granuloma formation in BCG-infected mice was abrogated, and the bacilli
multiplied in an uncontrolled manner, leading to decreased survival of
the animals (13). The protective role of TNF- was further
demonstrated in studies with mice in which the genes encoding TNF-
or the TNF- receptor 1 (TNF-R1) were disrupted. In TNF-R1
gene-disrupted (TNFR-KO) mice, infection with M. tuberculosis was not contained and the animals died soon after
infection (9). In TNF- gene-disrupted (TNF-KO) mice, M. tuberculosis infection led to dysregulated granuloma
formation, resulting in large accumulations of cells and mycobacteria
in the lungs, as well as extensive necrosis and neutrophil infiltration (1).
has been shown in many systems to
induce immunopathology in vivo. Tissue necrosis and cachexia or wasting
have been associated with elevated TNF- levels (3, 28).
In patients with tuberculosis, increases in this cytokine have been
implicated in clinical worsening (2). In a previous study in
mice, we showed that a reduction in TNF- levels in the infected lung
was associated with a decrease in granuloma size and less necrosis
(22). Although these results are suggestive of a pathogenic
role for this cytokine, it has been difficult to directly demonstrate
the deleterious effects of TNF- in murine tuberculosis.
in
the murine response to mycobacterial infection, we used a strain of
recombinant BCG that secretes murine TNF- to infect TNF-KO mice.
Some mice were given an unusually high inoculum of bacilli to increase
the amount of TNF- at the site of infection. Following intravenous
infection, we evaluated the growth of the recombinant BCG in the lungs,
livers, and spleens; the granulomatous response in the lungs and liver;
cytokine mRNA production in the lungs; and survival of the infected animals.
MATERIALS AND METHODS
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
p55 receptor gene-disrupted (TNFR-KO) mice on a
C57BL/6 genetic background, and homozygous TNF- gene-disrupted
(TNF-KO) mice on the same genetic background were used (19,
26). The mice were obtained from the University of Cape Town
breeding stock and were kept under specific-pathogen-free conditions at
the University of Cape Town animal facility until infection.
(BCG-TNF) or containing
the vector only (BCG-vector) were a kind gift from Richard Young of the
Whitehead Institute, Cambridge, Mass. Murine cDNA for TNF- was
cloned into the plasmids pRBD3 and pRBD4, as described previously
(24). The expression vectors contained a kanamycin
resistance gene. The BCG-TNF and BCG-vector cells were grown to mid-log
phase in Middlebrook 7H9 medium (Difco Laboratories, Detroit, Mich.)
containing kanamycin (18 µg/ml) (Sigma, St. Louis, Mo.) with minimal
agitation for 7 days and kept frozen in aliquots until use
(23).
80°C
until assay. Lungs, liver, and spleen were collected aseptically
immediately after cardiac puncture, weighed, and then used for
evaluation of bacillary load, cytokine mRNA expression levels, and
histology. Serum and organs from uninfected mice were used for
determination of baseline cytokine and cytokine mRNA levels.
Histology. Lungs and livers of mice were fixed in 10% buffered formalin, paraffin embedded, and processed for histology. Sections were stained with hematoxylin and eosin and Ziehl-Neelsen stain for histologic evaluation and photography.
Immunohistology. Formalin-fixed, paraffin-embedded sections were deparaffinized and rehydrated through graded alcohols. Antigen retrieval was accomplished by boiling the slides in 10 mM citrate buffer (pH 6.0) for 20 min. The staining of the sections was performed in an automated immunostainer (Ventana, Tucson, Ariz.) with a polyclonal rabbit anti-mouse antibody specific for inducible nitric oxide synthase (iNOS) (1:300) (Calbiochem, La Jolla, Calif.) (14).
PCR for cytokine mRNA.
Total cellular RNA was obtained from
lungs of mice at 14, 28, and 45 days following intravenous infection
with BCG-vector or BCG-TNF. Tissues were homogenized in 3 ml of RNAzol
B (Cinna/Biotcex Lab. Inc., Houston, Tex.) and RNA was extracted
according to the manufacturer's instructions. The reverse
transcription-PCR was carried out as previously described
(15). Briefly, 1 µg of RNA was reverse transcribed with a
Moloney murine leukemia virus reverse transcriptase and amplified with
Taq polymerase according to procedures given in the GeneAmp
RNA PCR kit (Perkin-Elmer, Branchburg, N.J.). Primers for TNF-,
interleukin 10 (IL-10), gamma interferon (IFN-
), IL-12, and
-actin were used as described previously (22).
Densitometry of the amplified bands was carried out with a
PhosphorImager (Molecular Dynamics, Sunnyvale, Calif.). Results were
normalized to the density of -actin.
CFU assay. Bacterial loads in the lungs, livers, and spleens of infected mice were evaluated by using 10-fold serial dilutions of organ homogenates. Organ homogenates were plated onto Middlebrook 7H10 agar plates (Difco) as well as 7H10 agar supplemented with 18 µg of kanamycin per ml (Sigma). The two sets of plates were incubated at 37°C for 3 weeks. Organisms were enumerated as CFU as described previously (22).
Determination of TNF- levels in plasma of infected mice.
At the time of sacrifice, mice were bled by cardiac puncture into
EDTA-containing tubes. Plasma was stored at 80°C until assay by
enzyme-linked immunosorbent assay (ELISA) for TNF- (Endogen, Inc.,
Boston, Mass.).
Determination of TNF- levels in the bacterial culture
supernatants.
The ability of the recombinant bacilli to secrete
murine TNF- was verified by measuring by ELISA (Endogen) the
concentration of the cytokine in the bacterial culture supernatants.
Only mycobacterial suspensions that produced the cytokine were used for
infection. To ensure that the recombinant BCG-TNF cells were still
secreting the cytokine during the infection, mycobacteria recovered
from the lungs of the infected mice (giving rise to colonies in the CFU
assay) were grown for 1 week in 7H9 medium supplemented with 18 µg of
kanamycin per ml, and the supernatants were tested for TNF-
concentrations by ELISA.
Statistical analysis. Data were analyzed with an independent t test when indicated. P values of <0.05 were considered statistically significant.
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RESULTS |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Effect of TNF- on growth of recombinant BCG in TNF-KO mice.
The effect of local production of TNF- on mycobacterial growth in
the organs of mice infected intravenously was evaluated. We compared
the bacillary load in the lungs, spleens, and livers of mice following
infection with about 105 (low dose) or about
107 (high dose) BCG-TNF cells to the bacillary load in mice
infected with the control BCG-vector. When the TNF-KO mice were
infected with BCG-vector, the infection was not controlled, and the
bacillary load increased in all organs at both doses of infection (Fig. 1). The increase in CFU count from
baseline to the 28-day time point of the study was significant at both
doses for all organs tested (P < 0.05). On the other
hand, when TNF-KO mice were infected with low-dose or high-dose
BCG-TNF, bacillary growth was controlled, and CFU decreased slightly
from baseline to the final time point, similar to the response seen in
wild-type mice (Fig. 1). By 28 days postinfection, there was a
significant difference in CFU between the BCG-TNF and BCG-vector in the
TNF-KO mice (P < 0.04).
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was detected in the plasma of any of the mice infected at
the low dose. However, this cytokine was detected at days 28 and 40 in
the plasma (10 to 13 pg of TNF- per ml) of TNF-KO mice infected with
the high dose of BCG-TNF.
Thus, in the TNF-KO mice, TNF- produced at the site of infection
appeared to reconstitute the host response, resulting in control of
bacterial growth even when the initial infecting inoculum was very high.
Effect of TNF- on lung histopathology following infection of
TNF-KO mice with recombinant BCG.
The levels of cellular
accumulation and organization (granulomatous response) in the lungs of
TNF-KO mice following infection with high or low doses of BCG-TNF or
BCG-vector were compared. At 28 days postinfection with about
105 organisms of either recombinant, the lungs of the
TNF-KO mice appeared relatively unaffected, with small scattered
cellular aggregates in the parenchyma. These aggregates consisted of
lymphocytes and macrophages, some of which stained for iNOS expression
(Fig. 2A and D). The iNOS-staining
macrophages appeared more focused in the BCG-TNF-infected lungs (Fig.
2A) than in BCG-vector infected lungs (Fig. 2D), which showed a
scattered pattern. By 45 days, the two infections differed markedly. In
mice infected with low-dose BCG-TNF, the granulomas remained small and
clearly distinct from the majority of the lung, which appeared normal
(Fig. 2B). These granulomas consisted of lymphocytes and macrophages
(Fig. 2C), some of which were still iNOS positive (not shown). In
contrast, in the TNF-KO mice infected with low-dose BCG-vector, the
granulomas had enlarged extensively and occupied most of the lung (Fig.
2E). These contained large undifferentiated macrophages, many
lymphocytes, and a few scattered polymorphonuclear leukocytes (Fig.
2F). iNOS staining was still evident (not shown). Infection of
wild-type mice with this low dose of BCG-TNF or BCG-vector resulted in
little or no granulomatous response (not shown).
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on the cellular
inflammatory response in the lungs, TNF-KO mice were infected with high
doses (107) of either BCG-TNF or BCG-vector. High doses of
infection induced aggressive and rapid responses. Already at 28 days
postinfection, extensive cellular recruitment into the lungs was noted
in response to either infection (Fig. 3A and
D). In mice infected with BCG-TNF, the
lung space surrounding the cellular aggregates was almost filled with
fluid and infiltrated with predominantly mononuclear cells and
scattered polymorphonuclear leukocytes (Fig. 3A to C). In the
BCG-vector infected mice, some lung tissue remained uninvolved with
intact air spaces (Fig. 3D to F). Staining of the lung sections for
acid-fast bacilli revealed fewer mycobacteria in mice infected with
BCG-TNF (Fig. 3C) than in mice infected with BCG-vector (Fig. 3F),
confirming the CFU data (Fig. 1). When wild-type mice were similarly
infected with high doses of BCG-TNF or BCG-vector, multiple small,
well-organized granulomas were seen in the lungs at 28 days, with few
if any acid-fast bacilli (Fig. 3G to I). In these animals, much more of
the lung airspace remained intact.
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in
the TNF-KO mice infected with BCG-vector led to uncontrolled cellular
recruitment into the infected lungs. On the other hand, the TNF-
secreted by the low dose of recombinant mycobacteria appeared to
regulate the granulomatous response in the TNF-KO mice, resulting in
smaller and better-differentiated granulomas by 45 days of infection.
However, high levels of TNF- (high enough to be detected in the
plasma) led to an overwhelming inflammatory response that compromised
lung function in the host, despite the successful control of the growth
of the infecting mycobacteria (Fig. 1).
Effect of TNF- on spleen weight in TNF-KO mice infected with
recombinant BCG.
Spleen weight, which may be used as an indicator
of the systemic immune response to infection, was monitored in the
gene-disrupted mice infected with either recombinant. At each time
point, the spleens recovered from mice were weighed. When TNF-KO mice
were infected with about 105 BCG-vector cells, there was a
dramatic increase in mean spleen weight from 0.065 mg to 0.395 mg
(P < 0.01) (Fig. 4). In
contrast, when TNF-KO mice were infected with 105 BCG-TNF
cells, there was less increase in the mean spleen weight, from 0.062 mg
to 0.215 mg (P < 0.03). From 28 days postinfection, a
statistically significant difference in spleen weight between infection
with BCG-TNF and infection with BCG-vector was noted (P < 0.01). At this time, the spleens from the TNF-KO mice infected with BCG-TNF were not much larger than those from wild-type mice infected with BCG-vector or BCG-TNF, which did not change in weight (Fig. 4).
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produced by the recombinant BCG
reduced the degree of spleen enlargement seen in the absence of TNF-
in the TNF- gene-disrupted mice. However, the presence of a larger
amount of TNF- appeared to exacerbate the inflammatory process,
leading to an increase in spleen weight despite the control of
bacillary growth (Fig. 1).
Effect of TNF- on survival of mice infected with recombinant
BCG.
The infection of TNF-KO mice with 5 × 106
BCG-vector cells resulted in early death of the mice (Fig.
5). By day 56, all animals had succumbed
to the infection. Following infection with 5 × 106
BCG-TNF cells, however, the TNF-KO mice survived (P < 0.001 for BCG-TNF versus BCG-vector). The wild-type mice infected
at this dose with BCG-vector or BCG-TNF had no deaths over the period of the experiment. Following infection of TNF-KO mice with high-dose (2 × 108) BCG-vector, 100% mortality had been
observed already by day 35 (P < 0.001 for TNF-KO
versus wild-type mice infected with BCG-vector) (Fig. 5). Infection
with high-dose (2 × 108) BCG-TNF resulted in similar
mortality (P = 0.4 for BCG-TNF versus BCG-vector in
TNF-KO mice), with about 70% of the TNF-KO mice succumbing by day 39 (P = 0.006 for TNF-KO mice infected with BCG-TNF versus
wild-type mice infected with BCG-vector) (Fig. 5). When wild-type mice
were infected with this dose of BCG-vector, 100% survived.
Interestingly, wild-type mice infected with this dose of BCG-TNF showed
some early mortality. By day 32, 33% of the mice had succumbed
(P = 0.06 for wild-type mice infected with BCG-TNF
versus those infected with BCG-vector). Therefore, it appears that the
TNF- from the recombinant mycobacteria restored the ability of the
TNF-KO mice to control the infection and survive. However, excess
levels of TNF- due to the presence of unusually large numbers of
recombinant BCG-TNF cells compromised the survival of the mice, whether
or not the growth of the bacilli was controlled (Fig. 1).
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Survival of TNF-R1-deficient mice infected with recombinant
BCG.
To determine whether the differences in survival of mice
required signaling by TNF-R1, TNFR-KO mice were infected
intravenously with either BCG-vector or BCG-TNF at a low (5 × 106) or a high (1 × 108) dose. There was
no difference in survival between the TNFR-KO mice infected with
BCG-vector and those infected with BCG-TNF at either dose (P > 0.1) (Fig. 6). This result
indicated that the absence of a functional TNF- signaling pathway
due to the lack of TNF- was the cause of the reduced survival
observed in the TNF-KO mice infected with BCG-vector (Fig. 5).
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Cytokine expression in tissues of TNF-KO mice infected with
recombinant BCG.
The early (14 day) expression of IFN-, IL-12,
and IL-10 mRNA in the lungs of TNF-KO-infected mice was studied. In
response to infection of TNF-KO mice with BCG-vector, relatively low
levels of IFN- mRNA were induced compared to the amount of IFN-
mRNA induced by infection with BCG-TNF (Table
1). The levels of IL-12 and IL-10 mRNA
expressed in the lungs were also lower in response to infection with
BCG-vector than those in response to BCG-TNF. By later time points, as
differences in the bacillary load became apparent, cytokine mRNA levels
increased in the lungs of mice infected with BCG-vector relative to the
lungs of mice infected with BCG-TNF (not shown). The results taken
together suggested that TNF- is required for the efficient
generation of the Th1-type (IFN- and IL-12) cellular immune response
to mycobacterial infection in mice.
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DISCUSSION |
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Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Here we directly demonstrate with the murine model of
mycobacterial infection that TNF- is not only essential for
protection, but at high levels is also seriously detrimental. We show
that when excess levels of the cytokine are present at the site of infection, as in the TNF-KO or wild-type mice infected with a high
inoculum of BCG-TNF, the toxic sequelae of excess TNF- override its
demonstrated protective effects, and the animals die in spite of
control of bacterial growth.
The protective role of TNF- in mycobacterial infections has been
well established in experimental models (1, 8-10, 13, 25).
The cytokine can exert a number of effects. TNF- has been shown to
modify the endothelium and induce chemokine expression, thereby
facilitating extravasation of monocytes and T cells from the blood and
directing this migration of cells to the infected site (13, 20,
32). In our previous studies, we showed that the local immune
response to mycobacterial antigens induces the expression of the CXC
chemokine IP10 (12), which is recognized by the T-cell and
NK-cell chemokine receptor CXCR3 (16, 17). The expression of
IP10 and other TNF--induced molecules by macrophages, fibroblasts,
and endothelial cells served to retain the activated T cells at the
site of infection (27). Indeed, in this study, we show that
TNF- is necessary for the accumulation and organization of
monocytes, macrophages, and lymphocytes into well-differentiated granulomas (Fig. 2). In addition, TNF- activates T cells or T-cell subsets, thereby facilitating the generation of cytokines and/or cytotoxic effector molecules and ensuring their sustained expression by
the cells (11, 30). TNF- also activates and fosters
differentiation of dendritic cells for enhanced IL-12 production and
antigen presentation (11). This effect is confirmed by our
results, which show levels of IFN- and IL-12 mRNA induced in the
BCG-TNF-infected lungs in the presence of TNF- higher than those
induced by the BCG-vector infection in the absence of this cytokine.
TNF- may also directly activate macrophages to control the growth of
and/or kill the intracellular mycobacteria (7).
TNF--activated murine macrophages can kill mycobacteria via the
generation of reactive nitrogen intermediates. It has previously been
shown that iNOS induction is associated with killing of mycobacteria in
vivo and in vitro (4, 18). However, our study reported here
shows that the presence of the iNOS protein in the macrophages of the
infected lungs is not sufficient to control the growth of the bacilli.
Tissue sections of the lungs and livers of the TNF-KO mice infected
with BCG-vector stained for iNOS (Fig. 2D), yet the infection was not
controlled (Fig. 1). Similar observations were recently made by Bean et
al. (1). Thus, it appears that in the absence of TNF-,
iNOS expression alone is not sufficient for the killing of
intracellular mycobacteria. Further aspects of TNF--mediated
macrophage activation and protection against mycobacteria are currently
being explored.
In addition to these protective effects, TNF- has detrimental
effects. When TNF- is present in large amounts systemically, the
resulting inflammatory cascade causes leaky capillaries, leukocyte infiltration, neutrophil-mediated endothelial damage, and inhibition of
pulmonary surfactant (28, 31). This damaging inflammation is
analogous to that seen in the lungs of adults with respiratory distress
syndrome (6, 14). TNF--mediated inflammatory damage is
also easily demonstrated in the central nervous system. For example, in
rabbits, intrathecal infection with virulent M. bovis strain
Ravenel results in local production of relatively high levels of TNF.
Excessive vasculitis, more extensive leukocyte migration into the
cerebrospinal fluid, and more severe pathology in the brain are
observed compared to inoculation with M. bovis strains that
induced less TNF (29). This pathologic inflammation in the
central nervous system is similar to that observed in patients with
tuberculous meningitis.
Our studies suggest that the extent of inflammation and cellular
recruitment to a site of mycobacterial infection are determined by a
number of independent factors. One of these is the load of infecting
bacilli. This is clearly demonstrated by the observation that
following infection of TNF-KO or TNFR-KO mice with the high dose of
BCG-vector, when no TNF- or TNF- activity is present, the
inflammatory cellular response was augmented and the mice died sooner
than following infection with the lower dose of BCG-vector. The other
determinant of inflammation is the level of TNF-. When TNF-KO mice
were infected with a high dose of BCG-TNF, the inflammatory response
was more severe than following infection with the same dose of
BCG-vector, and the mice died. Therefore, it appears that excessive inflammation, whether due to an unusually high level of
bacteria or of TNF-, leads to occupation of the air space by cells
and fluid, compromised lung function, and death of the animals.
Cytokines secreted by recombinant mycobacteria within the host cell
vacuoles have been shown to be biologically active. In a recent study,
infection of IFN- gene-disrupted mice with recombinant BCG secreting
murine IFN- (BCG-IFN-) resulted in reconstitution of the
protective immune response against BCG (21). In the same study, infection in vitro of peritoneal macrophages obtained from IFN- gene-disrupted mice with BCG-IFN- induced specific
activation of the nuclear factor STAT-1 and IFN regulatory factor 1 as
well as iNOS. Similarly, in the present study, the antimycobacterial immune response of TNF-KO mice was restored by infection with recombinant BCG secreting murine TNF-, and iNOS was induced in the
infected tissues. In vitro, we observed that macrophages obtained from TNF-KO mice infected with BCG-TNF are activated, as
demonstrated by induction of iNOS (G. Kaplan et al., unpublished data).
These results, taken together, demonstrate that the cytokines secreted by recombinant bacteria can be biologically active. However, the mechanisms by which the cytokines exert their effects, whether intracellular or extracellular, are not clear. The cytokines can be
measured in the culture supernatants of infected macrophages, as well
as in the plasma of mice infected at very high doses. This suggests
that, at least in some cases, active cytokine is secreted into the
extracellular milieu.
The protective and detrimental effects of TNF- in an infection have
also been similarly observed in a rodent model of malaria. In rats,
Plasmodium chaboudi infection cures spontaneously and TNF- is not detectable in the blood (5). Plasmodium
vinckei infection on the other hand, leads to death of infected
rats after several weeks. In the latter animals, high parasitemia
leading to high TNF- levels results in a shock-like condition and
focal liver necrosis. Although ultimately the parasitemia in this
infection is cleared, death of the animal from TNF--mediated shock
still ensues.
In summary, the results presented here show that TNF- is a
double-edged sword. While it is an essential component of the host
protective response against mycobacterial infection, high levels of the
cytokine at the site of infection induce an excessive inflammatory
response that overwhelms the beneficial effects of the cytokine.
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ACKNOWLEDGMENTS |
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We thank Victoria Freedman for help with preparation of the manuscript, Marguerite Nulty for secretarial assistance, and Judy Adams for preparation of the figures.
These studies were supported in part by Direct Effect (New York, N.Y.) and by AI42056 (G.K.). L.G.B. was a Fogarty International Fellow (AITRP TW00231 to Columbia University, New York, N.Y.).
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FOOTNOTES |
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* Corresponding author. Mailing address: The Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-8375. Fax: (212) 327-8376. E-mail: kaplang{at}rockvax.rockefeller.edu.
Present address: Infectious Diseases Clinical Research Unit, The
Lung Institute, University of Cape Town, Cape Town, South Africa.
Editor: S. H. E. Kaufmann
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REFERENCES |
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Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
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| 1. |
Bean, A. G.,
D. R. Roach,
H. Briscoe,
M. P. France,
H. Korner,
J. D. Sedgwick, and W. J. Britton.
1999.
Structural deficiencies in granuloma formation in TNF gene-targeted mice underlie the heightened susceptibility to aerosol Mycobacterium tuberculosis infection, which is not compensated for by lymphotoxin.
J. Immunol.
162:3504-3511 |
| 2. |
Bekker, L. G.,
G. Maartens,
L. Steyn, and G. Kaplan.
1998.
Selective increase in plasma TNF- and concomitant clinical deterioration after initiating therapy in patients with severe tuberculosis.
J. Infect. Dis.
178:580-584[Medline].
|
| 3. | Beutler, B., and A. Cerami. 1988. Tumor necrosis, cachexia, shock, and inflammation: a common mediator. Annu. Rev. Biochem. 57:505-518[CrossRef][Medline]. |
| 4. |
Britton, W. J.,
N. Meadows,
D. A. Rathjen,
D. R. Roach, and H. Briscoe.
1998.
A tumor necrosis factor mimetic peptide activates a murine macrophage cell line to inhibit mycobacterial growth in a nitric oxide-dependent fashion.
Infect. Immun.
66:2122-2127 |
| 5. | Clark, I. A., W. B. Cowden, G. A. Butcher, and N. H. Hunt. 1987. Possible roles of tumor necrosis factor in the pathology of malaria. Am. J. Pathol. 129:192-199[Abstract]. |
| 6. | Contreras, M., N. Hariharan, J. R. Lewandoski, W. Ciesielski, R. Koscik, and J. J. Zimmerman. 1996. Bronchoalveolar oxyradical inflammatory elements herald bronchopulmonary dysplasia. Crit. Care Med. 24:29-37[CrossRef][Medline]. |
| 7. | Denis, M., E. O. Gregg, and E. Ghandirian. 1990. Cytokine modulation of Mycobacterium tuberculosis growth in human macrophages. Int. J. Immunopharmacol. 12:721-727[CrossRef][Medline]. |
| 8. | Denis, M. 1991. Involvement of cytokines in determining resistance and acquired immunity in murine tuberculosis. J. Leukoc. Biol. 50:495-501[Abstract]. |
| 9. | Flynn, J. L., M. M. Goldstein, J. Chan, K. J. Triebold, K. Pfeffer, C. J. Lowenstein, R. Schreiber, T. W. Mak, and B. R. Bloom. 1995. Tumor necrosis factor-alpha is required in the protective immune response against Mycobacterium tuberculosis in mice. Immunity 2:561-572[CrossRef][Medline]. |
| 10. |
Garcia, I.,
Y. Miyazaki,
G. Marchal,
W. Lesslauer, and P. Vassalli.
1997.
High sensitivity of transgenic mice expressing soluble TNFR1 fusion protein to mycobacterial infections: synergistic action of TNF and IFN- in the differentiation of protective granulomas.
Eur. J. Immunol.
27:3182-3190[Medline].
|
| 11. |
Josien, R.,
B. R. Wong,
H. L. Li,
R. M. Steinman, and Y. Choi.
1999.
TRANCE, a TNF family member, is differentially expressed on T cell subsets and induces cytokine production in dendritic cells.
J. Immunol.
162:2562-2568 |
| 12. |
Kaplan, G.,
A. D. Luster,
G. Hancock, and Z. A. Cohn.
1987.
The expression of a gamma interferon-induced protein (IP-10) in delayed immune responses in human skin.
J. Exp. Med.
166:1098-1108 |
| 13. | Kindler, V., A.-P. Sappino, G. E. Grau, P. F. Pignet, and P. Vassalli. 1989. The inducing role of tumour necrosis factor in the development of bactericidal granulomas during BCG infection. Cell 56:731-740[CrossRef][Medline]. |
| 14. |
Kristof, A. S.,
P. Goldberg,
V. Laubach, and S. N. Hussain.
1998.
Role of inducible nitric oxide synthase in endotoxin-induced acute lung injury.
Am. J. Respir. Crit. Care Med.
158:1883-1889 |
| 15. | Laochumroonvorapong, P., J. Wang, C.-C. Liu, W. Ye, A. L. Moreira, K. B. Elkon, V. H. Freedman, and G. Kaplan. 1997. Perforin, a cytotoxic molecule which mediates cell necrosis, is not required for the early control of mycobacterial infection in mice. Infect. Immun. 65:127-132[Abstract]. |
| 16. |
Loetscher, M.,
B. Gerber,
P. Loetscher,
S. A. Jones,
L. Piali,
I. Clark-Lewis,
M. Baggiolini, and B. Moser.
1996.
Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes.
J. Exp. Med.
184:963-969 |
| 17. |
Luster, A. D.,
S. M. Greenberg, and P. Leder.
1995.
The IP-10 chemokine binds to a specific cell surface heparan sulfate site shared with platelet factor 4 and inhibits endothelial cell proliferation.
J. Exp. Med.
182:219-231 |
| 18. |
MacMicking, J. D.,
R. J. North,
R. LaCourse,
J. S. Mudgett,
S. K. Shah, and C. F. Nathan.
1997.
Identification of nitric oxide synthase as a protective locus against tuberculosis.
Proc. Natl. Acad. Sci. USA
94:5243-5248 |
| 19. |
Marino, M. W.,
A. Dunn,
D. Grail,
M. Inglese,
Y. Noguchi,
E. Richards,
A. Jungbluth,
H. Wada,
M. Moore,
B. Williamson,
S. Basu, and L. J. Old.
1997.
Characterization of tumor necrosis factor-deficient mice.
Proc. Natl. Acad. Sci. USA
94:8093-8098 |
| 20. | Ming, W. J., L. Bersani, and A. Mantovani. 1987. Tumor necrosis factor is chemotactic for monocyte and polymorphonuclear leukocytes. J. Immunol. 138:1469-1474[Abstract]. |
| 21. |
Moreira, A. L.,
L. Tsenova,
P. J. Murray,
S. Freeman,
A. Bergtold,
L. Chiriboga, and G. Kaplan.
2000.
Aerosol infection of mice with recombinant BCG secreting murine IFN- reconstitutes local protective immunity.
Microb. Pathog.
29:175-185[CrossRef][Medline].
|
| 22. | Moreira, A. L., L. Tsenova-Berkova, J. Wang, P. Laochumroonvorapong, S. Freeman, V. H. Freedman, and G. Kaplan. 1997. Effect of cytokine modulation by thalidomide on the granulomatous response in murine tuberculosis. Tuber. Lung Dis. 78:47-55[CrossRef][Medline]. |
| 23. |
Murray, P. J.,
A. Aldovini, and R. A. Young.
1996.
Manipulation and potentiation of antimycobacterial immunity using recombinant bacille Calmette-Guerin strains that secrete cytokines.
Proc. Natl. Acad. Sci. USA
93:934-939 |
| 24. |
O'Donnell, M. A.,
A. Aldovini,
R. B. Duda,
H. Yang,
A. Szilvasi,
R. A. Young, and W. C. DeWolf.
1994.
Recombinant Mycobacterium bovis BCG secreting functional interleukin-2 enhances gamma interferon production by splenocytes.
Infect. Immun.
62:2508-2514 |
| 25. |
Roach, D. R.,
H. Briscoe,
K. Baumgart,
D. A. Rathjen, and W. J. Britton.
1999.
Tumor necrosis factor (TNF) and a TNF-mimetic peptide modulate the granulomatous response to Mycobacterium bovis BCG infection in vivo.
Infect. Immun.
67:5473-5476 |
| 26. | Rothe, J., W. Lesslauer, H. Lotscher, Y. Lang, P. Koebel, F. Kontgen, A. Althage, R. Zinkernagel, M. Steinmetz, and H. Bluethmann. 1993. Mice lacking the tumour necrosis factor receptor 1 are resistant to TNF-mediated toxicity but highly susceptible to infection by Listeria monocytogenes. Nature 364:798-802[CrossRef][Medline]. |
| 27. |
Taub, D. D.,
D. L. Longo, and W. J. Murphy.
1996.
Human interferon-inducible protein-120 induces mononuclear cell infiltration in mice and promotes the migration of human T lymphocytes into the peripheral tissues and human peripheral blood lymphocytes-SCID mice.
Blood
87:1423-1431 |
| 28. | Tracey, K. J., and A. Cerami. 1992. Tumor necrosis factor and regulation of metabolism in infection: role of systemic versus tissue levels. Proc. Soc. Exp. Biol. Med. 200:233-239[CrossRef][Medline]. |
| 29. |
Tsenova, L.,
A. Bergtold,
V. H. Freedman,
R. A. Young, and G. Kaplan.
1999.
Tumor necrosis factor alpha is a determinant of pathogenesis and disease progression in mycobacterial infection in the central nervous system.
Proc. Natl. Acad. Sci. USA
96:5657-5662 |
| 30. | Ueta, C., H. Kawasumi, H. Fujiwara, T. Miyagawa, H. Kida, Y. Ohmoto, S. Kishimoto, and I. Tsuyuguchi. 1996. Interleukin-12 activates human gamma delta T cells: synergistic effect of tumor necrosis factor-alpha. Eur. J. Immunol. 26:3066-3073[Medline]. |
| 31. | Vassalli, P. 1992. The pathophysiology of tumor necrosis factors. Annu. Rev. Immunol. 10:411-452[CrossRef][Medline]. |
| 32. | Wuyts, A., P. Proost, and J. VanDamme. 1998. Interleukin-8 and other CXC-chemokines, p. 271-311. In A. Thomson (ed.), The cytokine handbook. Academic Press, New York, N.Y. |
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