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Infection and Immunity, April 1999, p. 1929-1934, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
CD40 Ligation Prevents Trypanosoma cruzi
Infection through Interleukin-12 Upregulation
Damien
Chaussabel,1
Frédérique
Jacobs,1
Jan
de Jonge,2
Marijke
de
Veerman,2
Yves
Carlier,3
Kris
Thielemans,2
Michel
Goldman,1 and
Bernard
Vray1,*
Laboratoire d'Immunologie
Expérimentale1 and Laboratoire de
Parasitologie,3 Faculté de Médecine,
Université Libre de Bruxelles, and Laboratorium
Fysiologie, Faculteit Geneeskunde, Vrije Universiteit
Brussel,2 Brussels, Belgium
Received 14 September 1998/Returned for modification 19 October
1998/Accepted 21 January 1999
 |
ABSTRACT |
Because of the critical role of the CD40-CD40 ligand (CD40L)
pathway in the induction and effector phases of immune responses, we
investigated the effects of CD40 ligation on the control of Trypanosoma cruzi infection. First, we observed that
supernatants of murine spleen cells stimulated by CD40L-transfected 3T3
fibroblasts (3T3-CD40L transfectants) prevent the infection of mouse
peritoneal macrophages (MPM) by T. cruzi. This phenomenon
depends on de novo production of nitric oxide (NO) as it is prevented
by the addition of N-nitro-L-arginine methyl
ester, a NO synthase inhibitor. NO production requires interleukin
(IL)-12-mediated gamma interferon (IFN-
) and tumor necrosis factor
alpha (TNF-
) synthesis as demonstrated by inhibition experiments
using neutralizing anti-IL-12, anti-IFN-, and anti-TNF-
monoclonal antibodies (MAb). We found that an activating anti-CD40 MAb
also directly stimulates IFN--activated MPM to produce NO and
thereby to control T. cruzi infection. To determine the in
vivo relevance of these in vitro findings, mice were injected with
3T3-CD40L transfectants or 3T3 control fibroblasts at the time of
T. cruzi inoculation. We observed that in vivo CD40
ligation dramatically reduced both parasitemia and the mortality rate
of T. cruzi-infected mice. A reduced parasitemia was still
observed when the injection of 3T3-CD40L transfectants was delayed 8 days postinfection. It was abolished by injection of anti-IL-12 MAb. Taken together, these data establish that CD40 ligation facilitates the
control of T. cruzi infection through a cascade involving IL-12, IFN-, and NO.
| |
INTRODUCTION |
CD40 is a cell surface receptor
expressed by various cells (B lymphocytes, dendritic cells,
hematopoietic progenitors, endothelial cells, and epithelial cells)
including monocytes and macrophages (56). Interaction of
CD40 with its CD40 ligand (CD40L) (4, 22) triggers a
pleiotropic pathway involved in both humoral and cellular immunity. By
exerting potent biological activities on CD4+ T cells and
antigen-presenting cells such as dendritic cells and macrophages
(49), this pathway plays a major role in anti-infective host
defense (21). Indeed, CD40-CD40L interactions result in the
secretion of multiple cytokines such as interleukin (IL)-1, IL-6, IL-8,
IL-10, IL-12, gamma interferon (IFN-), and tumor necrosis factor
alpha (TNF-) by immunocompetent cells. In particular, IL-12 has
emerged as a potent immunoregulatory cytokine involved in the control
of intracellular infections (44, 54, 55).
Trypanosoma cruzi is a hemoflagellate protozoan parasite
with intracellular multiplication. It infects humans as well as
domestic and wild mammals and is the etiological agent of Chagas'
disease (51). Experimental infection of BALB/c mice mimics
the human disease and allows the study of host defense mechanisms. It
displays an acute phase characterized by high parasitemia, followed by a chronic phase during which parasites become undetectable in peripheral blood while persisting in tissues. Various cytokines are
implicated in the control of T. cruzi infection in mice
including IL-12, IFN-, and TNF- (1-3, 8, 25, 42, 45, 47,
53). IFN- and TNF- have been shown to induce nitric oxide
(NO) synthesis (13, 33), which in turn plays a crucial role
in the control of T. cruzi infection in mice both in vitro
and in vivo (1, 19, 24, 34, 35, 37-40, 57). The present
work was undertaken to analyze the effect of CD40 ligation on cell
infection in vitro and to investigate whether a CD40L stimulation was
able to protect mice against T. cruzi infection.
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MATERIALS AND METHODS |
SC and mouse peritoneal macrophages.
Spleens were harvested
from male BALB/c mice (6 to 8 weeks old) purchased from Bantin & Kingman Universal (Hull, United Kingdom) and maintained in our animal
facilities on standard laboratory chow. Suspensions of erythrocyte-free
spleen cells (SC) were obtained by spleen dilaceration and treatment
for 30 s with distilled sterile water. SC (107
cells/ml) were then suspended in RPMI 1640 medium (GIBCO, Grand Island,
N.Y.). They were plated in a 24-well cell culture plate (Nunc,
Roskilde, Denmark) and incubated in a 5% CO2 and
water-saturated atmosphere.
Mouse peritoneal macrophages (MPM) were harvested from male BALB/c mice
by washing their peritoneal cavities with chilled Hank's balanced salt
solution without Ca2+ and Mg2+ (pH 7.4; GIBCO)
(38). They were allowed to adhere (2 × 105
cells/well) on round sterile coverslips (Thermanox, 13-mm diameter; Miles Scientific, Naperville, Ill.) in 24-well microplates for 2 h
at 37°C in a 5% CO2 atmosphere. Nonadherent cells were
removed by washing.
SC and MPM were cultured in RPMI 1640 medium supplemented with
N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic
acid (HEPES) (25 mM), glutamine (25 mM), fetal calf serum (10%),
penicillin (100 IU/ml), and streptomycin (100 µg/ml; GIBCO).
Infection of MPM and mice with T. cruzi.
For in vitro
experiments, T. cruzi trypomastigotes (Tehuantepec strain)
were obtained from infected fibroblasts as previously described
(14). Trypomastigotes (106 parasites/well) were
added to MPM in a 5:1 parasite-to-cell ratio. After 24 h, cultures
were washed to remove free parasites, and MPM were further incubated
for 24 h. Then, cells were fixed with methanol and stained with
Giemsa stain. The percentage of infected MPM and the mean number of
amastigotes per infected MPM were recorded after microscopical
examination of at least 200 cells per well. A parasitic index was
calculated by multiplying the percentage of infected MPM times the mean
number of amastigotes per infected MPM (58).
For in vivo experiments, BALB/c mice were inoculated intraperitoneally
with 100 blood-form trypomastigotes in 0.2 ml of Alsever's
solution.
Parasitemia was monitored by counting trypomastigotes
in blood samples
collected by tail incision every 2 days and every
day around the peak
of parasitemia. Survival rates were determined
daily.
CD40L-transfected fibroblasts.
3T3 fibroblasts transfected
with the gene encoding CD40L (3T3-CD40L transfectants) were obtained as
follows. The 5' region of the mCD40L, including the Kozack sequence,
the leader sequence, and the first 136 codons, was amplified by reverse
transcriptase-PCR of mRNA derived from activated EL4 T cells. The
primers used for amplification were based on the published sequence of
the mCD40L cDNA (4). The 460-bp downstream region of mCD40L
was obtained by BamHI and HindIII digestion
of the pH
APr-1-neo-mCD40L-mCD8 plasmid (kindly provided by P. Lane, Basel Institute for Immunology, Basel, Switzerland)
(31). The complete mCD40L cDNA was then cloned into the
pCI-neo vector (Promega, Leiden, The Netherlands). Flow cytometry
analysis of pCI-neo-mCD40L-transfected COS-7 cells, by using a
biotinylated anti-mCD40L antibody (Pharmingen, San Diego, Calif.),
showed that mCD40L was correctly assembled and expressed on the cell
surface. Plasmid pCI-neo-mCD40L was used for the lipofection (LipoTaxi;
Stratagene, Westburg, The Netherlands) of NIH 3T3 cells (American Type
Culture Collection, Rockville, Md.). mCD40L-expressing cells were
selected on the basis of growth in the presence of G418 sulfate (2 mg/ml, final concentration; Alexis Corporation, San Diego, Calif.) and
flow cytometry. For a negative control in the different assay systems
used, a 3T3 cell line transfected with empty pCI vector DNA (3T3
control fibroblasts) was selected.
3T3-CD40L transfectants and 3T3 control fibroblasts were grown in
Dulbeco's modified Eagle medium (GIBCO) supplemented with
fetal calf
serum (5%), penicillin (100 IU/ml), and streptomycin
(100 µg/ml),
and for transfected cells, with G418 sulfate (2 mg/ml).
They were
harvested after trypsin-EDTA treatment (GIBCO) and irradiated
at 30 Gy
(Mark I-68A irradiator; J.L. Shepherd and Associates,
San Fernando,
Calif.) to prevent further cell replication. They
were used to induce
in vitro CD40 triggering on SC. 3T3-CD40L
transfectants (5 × 10
4) were seeded together with 5 × 10
6 SC
per well (24-well culture plate) in 500 µl of culture
medium.
For in vivo experiments, mice were intravenously injected with
10
6 3T3-CD40L transfectants in 150 µl of
phosphate-buffered saline
(PBS). 3T3 control fibroblasts and PBS were
used as controls.
To assess the kinetics of IL-12 secretion, blood
samples (100
µl/mouse) were taken from uninfected mice by
retroorbital puncture
on days 0, 1, 4, 7, and 10 postinjection.
Individual blood samples
were centrifuged (10 min, 800 ×
g), and plasma samples were stored
at
70°C until use.
For in vivo neutralization of IL-12, anti-IL-12
monoclonal antibody
(MAb) (see below, 1.3 mg) was injected intraperitoneally
on the day of
infection (day 0) and then on days 2 and 4 postinfection
(p.i.).
MAbs and reagents.
Neutralizing anti-IFN- MAb (R4-6A2;
immunoglobulin G1 [IgG1]) and anti-TNF- MAb (MP6-XT3; IgG1) were
purchased from Pharmingen. The anti-IL-12 MAb (C17.8; IgG2a) used in
the in vitro experiments was purchased from Genzyme (Cambridge, Mass.).
Corresponding control isotype-matched antibodies R3-34 (IgG1) and
R35-95 (IgG2a) were purchased from Pharmingen. The hybridoma FGK45,
producing an IgG2a
specific for murine CD40, was kindly provided by
A. Rolink (Basel Institute for Immunology, Basel, Switzerland)
(43). The hybridoma cells were cultured in standard
conditions in RPMI 1640 containing 1% bovine serum. The rat MAb was
purified by affinity chromatography with a mouse anti-rat kappa MAb
immobilized on Sepharose beads and 3.5 M MgCl2 as elution
buffer. Eluted antibodies were extensively dialyzed against PBS and
filter sterilized. A nonrelated rat MAb was purified similarly and used
as a negative control. Another anti-CD40 MAb (3/23; IgG2a) was from
Pharmingen. The in vitro working concentration for all MAbs was 10 µg/ml except for anti-CD40 MAb (20 µg/ml). For in vivo experiments,
ascitic MAb anti-IL-12 (C17.8; IgG2a) was used (kindly provided by V. Flamand, Université Libre de Bruxelles, Brussels, Belgium). The
ascitic IgG2a antibodies used in vivo as a control were a kind gift
from H. Bazin (Université Catholique de Louvain, Brussels,
Belgium). FGK45 anti-CD40 MAb was also tested in vivo for its ability
to protect mice against T. cruzi infection. For this
purpose, we used high doses of FGK45 compared with the one used by
others (46): eight mice were injected intravenously with 200 µg of FGK45 MAb at the time of T. cruzi inoculation, and
four of them received again 100 µg of MAb at days 1 and 4 p.i.
Two control groups of five mice were injected with isotype-matched
control MAb or PBS.
N-nitro-
L-arginine methyl ester (NAME, 5 mM;
Sigma Chemical Co., St. Louis, Mo.) was used as the competitive
inhibitor of
NO
synthase.
Recombinant murine IFN-
(rIFN-
; 10 U/ml) was kindly supplied by
A. Billiau and H. Herremans (Rega Institute, Leuven,
Belgium).
The concentration of endotoxin in all the reagents and media was below
80 pg/ml according to the colorimetric limulus amoebocyte
lysate assay
(detection limit, 1 pg/ml) (Coatest endotoxin; Chromogenix,
Mölndal,
Sweden).
Cytokine determinations and nitrite assay.
Enzyme-linked
immunosorbent assay (ELISA) kits were purchased from Genzyme for
determination of IL-12 (p40 and p70), IFN-, and TNF-. The lower
limits of detection of these assays were, respectively, 15, 30, 20 and
35 pg/ml. NO production by MPM was assayed by measuring nitrite, its
stable degradation product, by the Griess reaction (20).
Supernatants (50 µl) from cultured MPM were harvested after 24 h
and mixed with 50 µl of Griess solution (1% sulfanilamide, 0.1%
naphthylethylene diamine dihydrochloride, 2%
H3PO4). The absorbance was measured at 540 nm
in a microplate ELISA reader (Spectracount Microplate Photometer;
Packard, Meriden, Conn.). Sodium nitrite (NaNO2) diluted in
culture medium was used as a standard. The detection limit of the assay
was 2.5 µM.
| |
RESULTS |
3T3-CD40L transfectants induce T. cruzi infection
clearance through a NO-mediated IL-12-dependent pathway.
The
involvement of CD40 ligation in T. cruzi infection was first
tested in vitro by using a two-step procedure. As T. cruzi readily infects 3T3 fibroblasts (data not shown), SC were first cocultured with either 3T3-CD40L transfectants, 3T3 control
fibroblasts, or medium. Supernatants of these cocultures were then
added to MPM cultured at the time of T. cruzi addition. The
parasitic index was calculated on MPM, 48 h later. Supernatants
from CD40L-activated SC clearly improved control of T. cruzi
infection by MPM, whereas supernatants of SC alone or cocultured with
3T3 control fibroblasts did not exert a significant effect (Fig.
1A).
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FIG. 1.
Effect of supernatants from CD40L-activated SC cultures
on T. cruzi infection of MPM. Supernatants harvested from SC
(open bars), from SC incubated with 3T3 control fibroblasts (hatched
bars), or from CD40L-activated SC (black bars) were added to MPM at the
time of T. cruzi infection, in the presence or absence of
NAME, and the parasitic index (A) and nitrite levels (B) were measured.
Data are means ± standard deviations from three independent
experiments performed in duplicate. ***, P < 0.001 compared to 3T3 control fibroblasts (Student's t
test).
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|
In light of the well-known protective role of NO in
T. cruzi
infection, NO levels were determined in culture supernatants
of MPM.
Supernatants from CD40L-activated SC induced a strong
up-regulation of
NO production by MPM. By contrast, the NO level
was low when MPM were
incubated with supernatants from SC-3T3
control fibroblast cultures or
SC alone (Fig.
1B). The addition
of NAME to MPM at the time of SC
supernatant addition abolished
the NO overproduction as well as their
effect on the parasitic
index (Fig.
1).
To identify the causative agents responsible for the induction of NO
production by MPM, IFN-
and TNF-
levels were measured
in
supernatants from CD40L-activated SC. As expected, IFN-
and
TNF-
synthesis was induced by 3T3-CD40L transfectants (data not
shown).
Furthermore, the addition of anti-IFN-
and anti-TNF-
MAbs to
CD40L-activated SC inhibited NO production by MPM, demonstrating
that
NO production was dependent upon the presence of these two
cytokines
(Table
1).
IL-12 is a major component of a complex biochemical pathway inducing
IFN-
synthesis and leading to NO production by macrophages.
Accordingly, IL-12 p40 and IL-12 p70 (the bioactive heterodimer)
levels
were assayed in the supernatants from CD40L-activated SC
collected
after 48 h (Table
2). In contrast to
supernatants from
SC cultured in the presence of 3T3 control
fibroblasts or medium
alone, we found a significant increase of both
IL-12 p40 and p70
when SC had been cocultured with 3T3-CD40L
transfectants. The
addition of neutralizing anti-IL-12 MAb to the
culture of CD40L-activated
SC inhibited the production of IFN-
by SC
(Fig.
2A). The supernatants
also failed
to induce NO production by MPM (Fig.
2B) and lost
their clearing effect
against
T. cruzi infection (Fig.
2C).
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FIG. 2.
Effect of neutralizing anti-IL-12 MAb added to
CD40L-activated SC on their IFN- and NO production and parasitic
index of T. cruzi-infected MPM. SC were activated with
3T3-CD40L transfectants in the presence of anti-IL-12 MAb,
isotype-matched control, or medium for 48 h. IFN- levels were
measured in culture supernatants after 48 h (A). Then, culture
supernatants were transferred to T. cruzi-infected MPM.
Nitrite levels (B) and parasitic index (C) were measured after 24 and
48 h, respectively. Data are means ± standard deviations
from three independent experiments performed in duplicate. ***,
P < 0.001 compared to data obtained with culture
supernatants harvested from SC incubated with isotype-matched control
(Student's t test).
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|
Activating anti-CD40 MAb directly enhances parasite control by
IFN--activated macrophages.
We also triggered the CD40-CD40L
pathway by using agonistic anti-CD40 MAb (FGK45) to directly activate
MPM in vitro. MPM were infected with T. cruzi
trypomastigotes and treated with anti-CD40 MAb together with a
suboptimal concentration of 10 U of rIFN- ml. FGK45 anti-CD40 MAb
clearly augments NO production by IFN--activated MPM (Table
3). Similar results were found with
another anti-CD40 MAb (3/23, data not shown). This NO up-regulation
correlated with an improved control of T. cruzi infection,
which was blocked in the presence of NAME but still active in the
presence of neutralizing anti-IL-12 MAb (Table 3).
Injection of 3T3-CD40L transfectants protects mice against T. cruzi infection.
To evaluate in vivo the protective effect
of CD40 stimulation on T. cruzi infection, mice were
injected with 3T3-CD40L transfectants, 3T3 control fibroblasts, or PBS
at the same time of T. cruzi inoculation. The injection of
3T3-CD40L transfectants reduced the peak of parasitemia (Fig.
3A) and considerably increased the
survival rate of T. cruzi-infected mice (Fig. 3B). Most of
the mice (10 of 15; n = 3) survived the acute phase of
infection and entered the chronic phase. In contrast, only one of the
infected mice survived after 3 weeks when the groups were injected
either with 3T3 control fibroblasts (1 of 14; n = 3) or
PBS (1 of 14; n = 3). These data indicate that a single
injection of 3T3-CD40L transfectants protects most of the mice against
fatal infection. However, injection of 400 µg of activating anti-CD40
MAb did not protect mice against T. cruzi infection (data
not shown).
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FIG. 3.
Effect of injection of 3T3-CD40L transfectants in
T. cruzi-infected mice. Three groups of mice were inoculated
with T. cruzi and injected with either 3T3-CD40L
transfectants, 3T3 control fibroblasts, or PBS. Parasitemia (A) and
cumulative survival (B) are reported. Dotted lines represent the
parasitemia when less than 50% of mice were still alive. Data were
pooled from three independent experiments (n = 14 or 15 mice per group). For parasitemia, the difference between the
experimental and control groups was significant (P < 0.01, Mann-Whitney U test) for the period from day 15 to day
24 p.i. **, P < 0.002 ( 2
analysis).
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|
We also tested the ability of 3T3-CD40L transfectants to modify the
course of an established infection. For this purpose,
mice were
inoculated with
T. cruzi and injected with 3T3-CD40L
transfectants or 3T3 control fibroblasts 8 days later. Treatment
by
3T3-CD40L transfectants reduced the peak of parasitemia to
4.0 × 10
6 parasite/ml versus 9.2 × 10
6
parasites/ml in mice injected with control transfectants (
P <
0.05 between day 20 and 29 p.i., Mann-Whitney U test). In
parallel,
the lethality rate was slightly reduced (on day 41, 14% of
control
mice survived versus 40% of the treatment group); however,
this
difference did not reach significance (
2 analysis).
The protective effect of CD40 ligation in T. cruzi-infected mice is related to up-regulation of IL-12.
To
assess the role of IL-12 in the protective effect of CD40 ligation in
vivo, we first measured IL-12 p40 levels in the serum of
3T3-CD40L-injected mice. These experiments confirmed that CD40-CD40L interactions induce IL-12 synthesis in vivo (Fig.
4). We then evaluated the effect of IL-12
neutralization on the outcome of infection. For this, neutralizing
anti-IL-12 MAb (or its isotype-matched control) was coinjected with
3T3-CD40L transfectants in T. cruzi-infected mice. Mortality
follow-up showed that protection obtained with injection of 3T3-CD40L
transfectants was prevented by IL-12 neutralization but not by
injection of the isotype-matched control (Fig.
5). These data establish that
CD40L-mediated protection in vivo depends on IL-12 release.
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FIG. 4.
Kinetics of IL-12 synthesis in serum of 3T3-CD40L
transfectant-injected mice. IL-12 p40 levels were measured in blood
samples obtained from mice injected with 3T3-CD40L transfectants
(n = 10) (black bars) or 3T3 control fibroblasts
(n = 8) (open bars). Data are means ± standard
errors of the means. *, P < 0.05; ***, P < 0.01 compared to day 0 (Student's t test).
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FIG. 5.
Effect of injection of neutralizing anti-IL-12 MAb on
T. cruzi-infected and 3T3-CD40L transfectant-treated mice.
Mice (n = 10) were inoculated with T. cruzi
trypomastigotes and injected with 3T3-CD40L transfectants or 3T3
control fibroblasts. Neutralizing anti-IL-12 MAb or isotype-matched
control was injected intraperitoneally on the day of infection (day 0)
and then on days 2 and 4 p.i. Cumulative survival was reported.
*, P < 0.05 (2 analysis).
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| |
DISCUSSION |
In the present study, we demonstrate that CD40 ligation leads to
the control of T. cruzi infection through the induction of NO. Several studies have shown that CD40-CD40L interactions could result in NO production (32, 48, 50, 52). Our results indicate that this is achieved by inducing IL-12 production as well as
by direct stimulation of IFN--activated macrophages. In vitro, high
levels of IL-12, IFN-, and TNF- are produced by CD40L-activated
SC. Supernatants from these SC cultures stimulate MPM, which become
able to control T. cruzi infection through NO production.
This NO production is inhibited when neutralizing anti-IL-12,
anti-IFN-, or anti-TNF- MAbs are added to the SC cultures with
3T3-CD40L transfectants. These data confirm that CD40-CD40L
interactions among SC promote the synthesis of IL-12 (9, 27,
31), which in turn, induces IFN- secretion (10, 41, 44,
55). IFN- acts in synergy with TNF- to stimulate the
production of NO (17), resulting in parasite clearing.
Finally, CD40 ligation also directly stimulates IFN--activated MPM
to produce NO and thereby to control T. cruzi infection.
This cascade is probably operative in vivo as a single injection of
3T3-CD40L transfectants to mice at the time of T. cruzi
inoculation exerts a clear-cut protective effect which is mediated by
IL-12. We show also that a single injection of 3T3-CD40L transfectants
even 8 days p.i. is still able to reduce parasitemia. This is in line with previous studies showing that treatment of T. cruzi-infected mice with anti-IL-12 MAb has an exacerbating effect
on both parasitemia and mortality (3) while an exogenous
supply of IL-12 protects mice (25).
Our results are also consistent with a recent study showing an
IL-12-dependent protection in mice infected with Leishmania (a closely-related parasitic protozoa) and injected with
anti-CD40-stimulating MAb (15). Furthermore, exacerbated
Leishmania infection is observed when CD40-CD40L
interactions are disrupted (6, 23, 26, 48). Likewise, a
CD40-CD40L-mediated protective effect is also observed with other
pathogens such as Cryptosporidium parvum (12) and
Pneumocystis carinii (59). In contrast, this is
not the case with other pathogens such as Borrelia
burgdorferi (16), Listeria monocytogenes
(21), Mycobacterium tuberculosis (7), and Histoplasma capsulatum (60). This discrepancy
could be explained by differential abilities of infectious agents to
induce IL-12 production by the host (7, 60). CD40-CD40L
interaction would be determinant only when IL-12 production induced by
pathogens is insufficient, as has been shown in the course of
Leishmania infection (5).
3T3-CD40L transfectants are found to be more efficient than activating
anti-CD40 MAb in improving parasite control, and this data is in
agreement with previous observations obtained with B cells
(30). This is most likely due to a more efficient
cross-linking of CD40 molecules by CD40L expressed at high density at
the fibroblast membrane compared with the agonistic anti-CD40 IgG MAb.
CD40L stimulating properties have already been used in the treatment of
tumors, tumor regression being linked to restoration of major
histocompatibility complex class I expression by tumor cells (18,
28, 36), IL-12 overproduction, or potentiation of host
antigen-presenting cell functions. Moreover, CD40 engagement restores,
at least in vitro, production of IL-12 by cells from human
immunodeficiency virus (HIV)-infected patients and it stimulates macrophages to produce HIV-1-suppressive chemokines (11,
29). According to our results, activation of the immune system
through CD40 ligation could also be considered a potent strategy for
immunotherapy of parasitic diseases.
| |
ACKNOWLEDGMENTS |
We thank H. Herremans and A. Billiau (Rega Institute, Leuven,
Belgium) for providing rIFN-; A. Scheich, who edited the English text; V. Vercruysse for valuable technical assistance; and I. Mazza for
help in preparing the manuscript.
This work was supported by grants from Action de Recherche
Concertée de la Communauté Française de Belgique, the
Fonds Emile Defay, the Sportvereniging tegen Kanker (M.d.V.), and the Fund for Scientific Research-Flanders.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Laboratoire
d'Immunologie Expérimentale (CP 615), Faculté de
Médecine, Université Libre de Bruxelles, route de Lennik,
B-1070 Brussels, Belgium. Phone: 32-2-555.62.60. Fax: 32-2-555.63.60. E-mail: bvray{at}med.ulb.ac.be.
Editor:
S. H. E. Kaufmann
| |
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Infection and Immunity, April 1999, p. 1929-1934, Vol. 67, No. 4
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Copyright © 1999, American Society for Microbiology. All rights reserved.
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Abstract
Introduction
