| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
World Health Organization Immunology Research and Training Center, Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
,
even when IL-4 has been produced for 48 h, resulted in maintenance
of IL-12R ß2-chain mRNA expression and IL-12 responsiveness. The data
presented here support the hypothesis that the genetically determined
susceptibility of BALB/c mice to infection with L. major
is primarily based on an up-regulation of IL-4 production, which
secondarily induces extinction of IL-12
signaling. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The initial demonstration of the dominant role of cytokines during the
early stage of antigenic stimulation in directing the functional
differentiation of CD4+ T cell precursors from TCR 
ß
transgenic mice in vitro (2, 3, 4) has been largely confirmed in vivo
using the murine model of infection with L. major. Thus,
during the initial period of infection, IL-12 and IL-4 have been
demonstrated to play critical roles in mediating the differentiation of
Th1 and Th2 subsets as well as the ability to contain or the failure to
contain lesion development, respectively (5, 6, 7, 8).
The aberrant Th2 response developing in BALB/c mice after infection with L. major, resulting in progressive disease, critically depends on the production of IL-4 early after infection. This contention is supported by previous studies from this laboratory, which have shown that BALB/c mice, in contrast to mice from resistant strains, exhibit a burst of IL-4 transcripts in CD4+ T cells in draining lymph nodes within 16 h of infection (9). This burst of IL-4 occurred during the period in which neutralizing IL-4 redirected protective Th1 cell development in BALB/c mice (8).
Comparing the intrinsic tendencies of CD4+ T cells from TCR
ß transgenic mice from different genetic backgrounds to mature
into either Th1 or Th2 effector cells in vitro, naive CD4+
T cells from BALB/c mice were shown to preferentially differentiate
toward the Th2 phenotype after priming under otherwise neutral
conditions, i.e., in the absence of exogenous cytokines (10). This
biased Th2 cell development has been recently proposed to proceed from
a rapid loss of responsiveness to IL-12 by BALB/c CD4+ T
cells during priming in vitro (11). The IL-12R comprises two
components, the IL-12R ß1 and the IL-12R ß2 subunits (12), which
are expressed only on T cells having engaged their TCR (13). Recent
data have clearly shown that extinction of IL-12 signaling during
priming in vitro of naive CD4+ T cells from TCR ß
transgenic BALB/c mice results from a selective loss of expression the
IL-12R ß2-chain (13), which is necessary for IL-12 signaling through
the JAK/STAT pathway (14). The presence of IL-4 during priming in vitro
strongly inhibited the expression of the IL-12R ß2-chain by activated
T cells, an effect that was over-riden by IFN- (13).
In this context we have recently shown that from 48 h after
infection with L. major, administration of exogenous IL-12
to BALB/c mice becomes ineffective in preventing the development of a
Th2 response (15). At this time, CD4+ T cells from infected
BALB/c mice no longer responded to IL-12 in terms of IFN- production
in vitro. Based on these findings, we initiated studies aimed at
analyzing the molecular basis of this state of unresponsiveness to
IL-12 rapidly induced in vivo following infection of BALB/c mice with
L. major and its regulation by other cytokines.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Female BALB/c and C57BL/6 mice were purchased from IFFA Credo
(St. Germain sur l’Abresle, France). BALB/c mice homozygous for
disrupted IFN-R -chain gene (IFN-R-/-) (16) were
derived from IFN-R-/- mice on the Sv129 genetic
background (17) by seven serial backcrosses to BALB/c mice. All mice
were used at 6–8 wk of age.
Parasites, infection, and treatment of mice
L. major LV 39 (MRHO/Sv/59/P strain) were maintained in vivo and grown in vitro as previously described (18). Mice were infected s.c. in the hind footpads with 3 x 106 stationary phase L. major promastigotes in a final volume of 50 µl. At various times following infection, mice were sacrificed, and their popliteal lymph nodes were removed for further analysis.
Recombinant murine IL-12 was provided by Hoffmann-La Roche (Nutley,
NJ). Recombinant IFN- was provided by Genentech (South San
Francisco, CA). BALB/c mice were treated i.p. with one injection of 1
µg of IL-12 or 105 U of IFN- at various times with
regard to infection. Anti-murine IL-4 and anti-murine IFN- mAb
were produced from the 11B11 rat hybridoma (19) or the rat XMG 1.2
hybridoma (20), respectively, and purified as previously described
(15). Mice were treated with 1 mg of the corresponding Ab at various
time points with regard to infection.
RNA extraction, and competitive and qualitative PCR
Total RNA was isolated from popliteal lymph node cells as previously described (21). First-strand cDNA synthesis was performed on total RNA using a first-strand cDNA synthesis kit (Pharmacia, Uppsala, Sweden). The semiquantitative PCR developed by Reiner et al. (22) was performed using the competitor construct PQRS containing sequences for multiple cytokines. Primer sequences (hypoxanthine guanine phosphoribosyl transferase (HPRT)4 and IL-4) and PCR conditions were used as described by the authors. The first-strand cDNA was used directly as a template in the presence of fivefold serial dilutions of the competitor plasmid. After separation of the PCR products by electrophoresis in agarose gel containing ethidium bromide, the ratio of the relative concentration of the IL-4 gene to the relative concentration of HPRT was calculated (22). Results are expressed as the fold increase in mRNA expression in mice infected with L. major compared with that in noninfected mice.
Expression of IL-12R ß1- and ß2-chain mRNA was monitored using a qualitative PCR assay. HPRT levels for all samples of a given experiment were assessed by the PCR method described by Reiner et al. (22). All samples were normalized with respect to their HPRT content. Using normalized samples and control plasmids containing the respective cDNA (12, 23), IL-12R ß1- and ß2-specific PCR reactions were performed using the following primers: ß1 3', 5'-ATT CTT GGG GTT CTT GGA GGC-3'; ß1 5', 5'-GCC AAG ATT AAG TTC TTG GTG-3'; ß2 3', 5'-GCG TCG GTA CTG AAT TTC GCA-3'; and ß2 5', 5'-GGG AGT ACA TAG TGG AAT GGA-3'. The PCR conditions used for the amplification of the IL-12R ß1- and ß2-chain cDNA were similar to those used for HPRT cDNA, except for the annealing temperature, which was 58°C. PCR products were visualized on 1% agarose gels stained with ethidium bromide and scored for expression or lack of expression. All reactions were repeated two or three times to ensure reproducibility.
Lymphocyte cultures and detection of IFN- in supernatants
CD4+ T cells were purified from the draining
popliteal lymph nodes by magnetic cell sorting (Miltenyi Biotech,
Bergish-Gladbach, Germany) according to the manufacturer’s procedure.
Cells (5 x 105) were stimulated with UV-irradiated
L. major promastigotes (1 x 106) in the
presence of 5 x 106 irradiated (3000 rad) spleen
cells from normal mice in the presence or the absence of IL-12 (10
ng/ml) in a final volume of 1 ml. Cells were cultured in DMEM
supplemented with 5% heat-inactivated FCS, L-glutamine
(216 mg/ml), 5 x 10-5 M 2-ME, and 10 mM HEPES in an
atmosphere of 7% CO2. Culture supernatants were collected
after 72 h of stimulation and were stored at -20°C until use.
IFN- was measured in supernatants by ELISA as previously described
(24). Mouse rIFN- (supernatant of L1210 cells transfected with the
murine IFN- gene, gift from Y. Wanabe, Kyoto University, Kyoto,
Japan) was used as standard. The limit of the detection of the assay
was 10 U/ml.
Statistics
Data were analyzed statistically by Student’s t test.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In a previous study we observed that from 48 h after
infection with L. major, treatment of BALB/c mice with
exogenous IL-12 was no longer capable of inhibiting the development of
a CD4+ Th2 response, reflected by increased IL-4 mRNA
expression from day 5 of infection, or enhancing IFN- mRNA
expression. This loss of responsiveness to IL-12 in vivo was
corroborated by results showing that 3 days after infection,
CD4+ T cells in draining lymph nodes of BALB/c mice were no
longer responsive to IL-12 in terms of IFN- production following
specific activation in vitro (15). This state of unresponsiveness was
prevented by neutralization of the IL-4 produced during the first days
of infection.
Experiments were designed to assess whether the unresponsiveness to
IL-12 rapidly induced in CD4+ T cells of BALB/c mice
following infection with L. major proceeded from a loss of
transcription of the IL-12R ß2-chain gene. The results in Fig. 1
confirm that 5 days after infection
with L. major, CD4+ T cells from draining lymph
nodes of BALB/c mice specifically stimulated with L. major
in vitro do not respond to IL-12 in terms of enhanced IFN-
production. RT-PCR analysis revealed that the IL-12R ß2 subunit mRNA
was completely absent in CD4+ lymph node T cells of BALB/c
mice 5 days after infection with L. major (Fig. 2). In contrast, similar analysis in
resistant C57BL/6 mice showed that CD4+ T cells were
responsive to IL-12 and expressed IL-12R ß2 mRNA at high levels. The
CD4+ T cells from BALB/c mice treated with anti-IL-4
mAbs at the initiation of infection also expressed high levels of the
IL-12R ß2 mRNA (Fig. 2) and produced increased amounts of IFN-
upon specific activation in vitro in the presence of IL-12 (Fig. 1).
|
|
). On days 2 and 5, although
IL-12R ß1 mRNA was maintained in CD4+ T cells from both
C57BL/6 and BALB/c mice, only C57BL/6 cells expressed IL-12R ß2 mRNA,
which was no longer detectable in BALB/c cells. Neither IL-12R ß1 nor
IL-12R ß2 mRNA was detectable in lymph node cells from noninfected
mice (Fig. 3). Similar results were obtained 7 days after infection
(data not shown).
|
Effect of treatment of BALB/c mice with exogenous IL-12 or IFN-
on maintenance of responsiveness to IL-12 and transcription of the
IL-12R ß2 subunit
It has been shown that the presence of IL-12 or IFN- during
priming in vitro of TCR ß transgenic BALB/c CD4+ T
cells resulted in maintenance of IL-12R ß2 subunit mRNA expression
and prevented the inhibition of IL-12 signaling, even when IL-4 was
also present in cultures (13). We therefore tested whether treatment of
BALB/c mice with exogenous IL-12 or IFN- at various times after
inoculation of parasites resulted in maintenance of both responsiveness
to IL-12 and IL-12R ß2 mRNA expression.
The results in Figs. 4 and
5 show that 5 days after infection,
CD4+ T cells from the draining lymph nodes of BALB/c mice
given IFN- at the initiation of infection or 16 or 48 h later
still responded to IL-12 in terms of enhanced production of IFN- in
vitro and expressed IL-12R ß2 subunit mRNA. Administration of 1 µg
of IL-12 to BALB/c mice at the time of infection or 16 h later
also resulted in maintenance of IL-12 responsiveness (Fig. 4) and
IL-12R ß2-chain mRNA expression (Fig. 5). In contrast, when given 2
days after initiation of infection, IL-12 was no longer able to either
reverse the IL-12 unresponsiveness of CD4+ T cells (Fig. 5)
or rescue the IL-12R ß2 subunit mRNA expression (Fig. 4). These
results show that IFN-, in contrast to IL-12, is effective in
maintaining IL-12 signaling in CD4+ T cells in vivo even if
administered when IL-4 has been produced during 48 h.
|
|
dependent
The effect of IL-12 on the differentiation of TCR ß
transgenic CD4+ T cells toward the Th1 phenotype in vitro
has been shown to require IFN- (25). Furthermore, we have recently
shown that exogenous IL-12 given to BALB/c mice during the first day of
infection significantly enhanced IFN- mRNA expression in draining
lymph node cells (15). Thus, it was relevant to assess whether the
capacity of treatment with IL-12 during the first day of infection to
maintain IL-12 responsiveness and IL-12R ß2 subunit mRNA expression
in draining lymph node CD4+ T cells was the consequence of
the IFN- produced in vivo.
The results in Fig. 6 confirm that
treatment of BALB/c mice with exogenous IL-12 during the first day of
infection results in maintenance of responsiveness to IL-12 of
specifically stimulated CD4+ T cells obtained 5 days after
infection. However, when the IFN- produced in vivo in response to
treatment with IL-12 was neutralized by administration of a specific
mAb, the CD4+ T cells became unresponsive to IL-12 in vitro
in terms of enhanced IFN- production (Fig. 6). The strict
correlation between the IL-12 responsiveness and the expression of
IL-12R ß2-chain mRNA is exemplified further by results showing that
the maintenance of IL-12R ß2-chain mRNA expression by
CD4+ T cells from BALB/c mice as a result of administration
of exogenous IL-12 during the first day of infection is abrogated
following neutralization of the IFN- produced in vivo (Fig. 7). To confirm the IFN- dependence of
the effect of treatment of BALB/c mice with IL-12 on the maintenance of
responsiveness to IL-12 by CD4+ T cells, this effect of
IL-12 was investigated in IFN-R -/- BALB/c mice.
|
|
clearly show that 5 days after infection,
CD4+ T cells from BALB/c IFN-R-/- mice,
treated or not with exogenous IL-12 at the initiation of infection, did
not produce increased amounts of IFN- when specifically stimulated
in vitro in the presence of IL-12. These mice developed a polarized Th2
response (data not shown). Inhibition of IL-12R ß2-chain mRNA expression pertains to the development of a polarized Th2 response in BALB/c mice infected with L. major
Loss of IL-12 responsiveness could represent an early step in the engagement of CD4+ T cells to the Th2 pathway of differentiation. Therefore, experiments were designed to assess, in BALB/c mice, whether the differentiation of disparate Th subsets following infection with L. major correlates with the maintenance or inhibition of expression of IL-12R ß2-chain mRNA in CD4+ T cells, leading to the maintenance or loss of IL-12 signaling, respectively.
Following the initial burst of IL-4 mRNA expression within 16 h in
CD4+ T cells in draining lymph nodes of BALB/c mice in
response to L. major, we have documented a second wave of
IL-4 transcripts in CD4+ T cells from day 5 of infection
onward, reflecting the development of a polarized Th2 response (15). We
have shown above that treatment of BALB/c mice with either exogenous
IFN- or IL-12 during the first day of infection resulted in the
maintenance of IL-12 responsiveness and IL-12R ß2-chain mRNA
expression in draining lymph node CD4+ T cells 5 days after
infection. Thus, we further studied the effects of these treatments on
the IL-4 mRNA expression normally seen in control infected BALB/c mice
on day 5. The results depicted in Fig. 8
show that administration of either IFN- or IL-12 to BALB/c mice at
the time of parasite inoculation or 16 h later resulted in a clear
inhibition of the IL-4 mRNA expression normally seen in
CD4+ T cells from control infected mice.
|
Abs (Fig. 5). Although
inhibition of IL-4 mRNA expression was still observed when IFN- was
given 48 h after infection, mice given IL-12 48 h after
infection exhibited increased IL-4 mRNA levels similar to those seen in
control infected mice (Fig. 8). Identical results were observed when
levels of IL-4 mRNA were determined 10 days after infection (data not
shown), indicating that these treatments did not simply delay IL-4 mRNA
expression. Together, these data show that 1) the loss of IL-12R
ß2-chain mRNA expression and IL-12 signaling in CD4+ T
cells from BALB/c mice infected with L. major correlates
with Th2 cell development; and b) the maintenance of IL-12R ß2-chain
mRNA expression and IL-12 signaling in vivo following administration of
exogenous cytokines is clearly related to an inhibition of Th2 cell
development. |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The expression of the IL-12R ß1-chain was maintained on
CD4+ cells from mice from both strains. The results
obtained demonstrate that maintenance or loss of IL-12R ß2-chain mRNA
expression by CD4+ T cells in vivo strictly correlates with
their responsiveness or lack of responsiveness to IL-12 in vitro,
respectively. Thus, expression of the IL-12R ß1-chain by itself does
not appear crucial for IL-12 signaling (13). Neither chain of the
IL-12R was expressed on CD4+ T cells from noninfected
C57BL/6 or BALB/c mice, indicating that in vivo TCR engagement is also
necessary for the expression of IL-12R. Together, these results
demonstrate the importance of the IL-12R ß2 subunit for IL-12
signaling in vivo. A similar conclusion was reached from the results of
experiments performed in vitro, using human and murine cell lines and
TCR ß transgenic CD4+ T cells (13, 26).
Down-regulation of IL-12R ß2-chain expression on CD4+ T cells from BALB/c mice was prevented by neutralization of the IL-4 produced as a result of the early IL-4 transcriptional burst previously shown to occur in draining lymph nodes within 16 h after the s.c. injection of L. major (9). Neutralization of this IL-4 at the initiation of infection has been demonstrated to be capable of redirecting protective Th1 cell development in BALB/c mice (8, 15). Furthermore, previous results from our laboratory have shown that the production of IL-4 is necessary only during a short period of time (<48 h after infection) for irreversible Th2 cell development to occur in BALB/c mice (15). Interestingly, we show here that the IL-4-mediated inhibition of the IL-12R ß2 chain expression in CD4+ T cells from BALB/c mice also occurs 48 h after infection. In contrast, CD4+ T cells from resistant C57BL/6 mice, in which Th1 cell maturation does occur, maintain IL-12R ß2-chain expression and IL-12 responsiveness at least up to the time when the functional differentiation of CD4+ T cells starts.
Together, these results strongly suggest that in vivo the maintenance or loss of IL-12R ß2-chain expression and the resulting responsiveness or lack of responsiveness to IL-12 represent early and important steps in the stable commitment of naive CD4+ T cells to the Th1 or Th2 pathway of differentiation, respectively. This conclusion is supported by our recent results, which have shown that the inhibition of Th2 cell development and the striking redirection of the resulting effector cells to the Th1 phenotype observed in BALB/c mice following treatment with anti-IL-4 mAb at the onset of infection with L. major require IL-12 (15). By losing the expression of the IL-12R ß2 subunit, CD4+ T cells induced by IL-4 to differentiate to the Th2 pathway rapidly become refractory to the Th1-differentiating signals of IL-12.
The observations reported here reinforce the critical importance of the
IL-4 produced in BALB/c mice during the first day of infection for the
subsequent differentiation of parasite-specific CD4+ T cell
precursors toward the Th2 phenotype. In this context we have recently
observed that the early burst of IL-4 mRNA expression seen in draining
lymph nodes of BALB/c mice within 16 h of infection occurs within
CD4+ T cells that express the Vß4 V8 TCR chains and
recognize the LACK Ag of L. major (27). This rapidly
produced IL-4 has an essential role in instructing subsequent Th2 cell
differentiation and consequently susceptibility to L. major,
since BALB/c mice lacking CD4+ T cells expressing the Vß4
TCR chain do not exhibit an early IL-4 response to L. major,
subsequently develop a Th1 response, and are fully resistant to
infection (27). Interestingly, CD4+ T cells from these
Vß4 CD4+ T cell-deficient BALB/c mice maintained their
expression of the IL-12R ß2-chain at least up to 8 days after
infection with L. major (H. Himmelrich et al., in
preparation).
These results strongly suggest that in the absence of IL-4,
differentiating CD4+ T cells maintain expression of the
IL-12R ß2 subunit and default to the Th1 pathway. Thus, it is
possible that the presently observed maintenance of the IL-12R
ß2-chain expression on CD4+ T cells from C57BL/6 mice
following infection with L. major results from the fact that
these mice do not produce sufficient amounts of IL-4 in response to
infection. It is noteworthy that we never observed a significant
increase in IL-4 mRNA expression in draining lymph nodes of C57BL/6
mice following infection with L. major (9). Similarly, the
preferential differentiation of TCR ß transgenic CD4+
T cells toward the Th2 phenotype when specifically primed in vitro in
the presence of IL-4 has been extensively demonstrated (4, 27). This
effect has been recently correlated with a selective loss of IL-12R
ß2-chain expression and IL-12 signaling (13). The genetic tendency of
naive CD4+ T cells from BALB/c mice to differentiate to the
Th2 pathway in vitro (10) has been proposed to be the consequence of a
loss of IL-12 signaling rather than an overproduction of IL-4 (11).
Nonetheless, neutralization of IL-4 with anti-IL-4 mAb during
priming in vitro of TCR ß transgenic CD4+ BALB/c T
cells resulted in maturation of a Th1 response (4), confirming the
critical role of the small amount of IL-4 produced during priming on
Th2 cell development. These data combined with our results showing that
the IL-4 rapidly produced in BALB/c mice in response to L.
major readily down-regulates the expression of the IL-12R ß2
subunit on CD4+ T cells, rather, suggest that the
genetically determined tendency to develop a Th2 response, resulting in
susceptibility to infection with L. major, is primarily
based on an up-regulation of IL-4 production that secondarily induces
extinction of IL-12 signaling.
The role of IL-4 in Th2 cell development and susceptibility of BALB/c mice to infection with L. major has been questioned by results that have shown that IL-4 knockout BALB/c mice neither heal their cutaneous lesions nor switch their CD4+ T cell response toward the Th1 phenotype (28). Even though other studies performed using BALB/c mice with homologous disruption of the IL-4 gene have led to conflicting results (29), it is possible that the commitment of CD4+ T cells, developing in the IL-4 free environment of IL-4 knockout mice, to the Th2 pathway is driven by a factor(s) different from the IL-4 normally directing the Th2 maturation of CD4+ T cells in wild-type BALB/c mice. This hypothesis deserves experimental testing.
Treatment of BALB/c mice with either IFN- or IL-12 at the initiation
of infection resulted in the maintenance of IL-12R ß2-chain
expression induced on CD4+ T cells following TCR engagement
and responsiveness to IL-12. This effect cannot simply be attributed to
the ability of these cytokines to down-regulate the early burst of IL-4
transcripts occurring in these mice following infection with L.
major (9), since exogenous IFN- was still effective in
maintaining IL-12R ß2-chain expression and IL-12 signaling even if
given when IL-4 had been produced for 48 h. The effect of
exogenous IL-12 was clearly the result of the IFN- produced in vivo
following this treatment, since it was abrogated by neutralization of
IFN-. Thus, the lack of effect of IL-12 when administered 48 h
after infection, i.e., at a time when CD4+ T cells from
infected BALB/c mice have lost expression of the IL-12R ß2 subunit,
probably results from its inability to signal and induce IFN-
production. These results imply that IL-12 by itself does not
up-regulate the expression of the ß2-chain of its receptor. They also
confirm previous data obtained in vitro (13), which have shown that
IFN- is capable of superseding the inhibition of IL-12R ß2-chain
expression caused by IL-4.
As already suggested, this effect of IFN- could explain the
requirement for this cytokine, observed in some experimental systems,
in the IL-12-generated development of Th1 cell in vitro (25). Indeed,
under conditions when IL-4 is produced during priming in vitro, the
presence of IFN-, by its ability to maintain IL-12R ß2 subunit
mRNA expression, will permit the IL-12 signaling necessary for Th1 cell
maturation (13). In this context, it has been shown that 3 days after
infection with L. major, lymph nodes from BALB/c mice
treated with IFN- produce, upon specific stimulation in vitro,
substantially less IL-4 and more IFN- than lymph nodes from
similarly infected mice not administered IFN- (30). Furthermore,
treatment with IFN- of BALB/c mice infected with L.
major, although not significantly altering the course of disease,
results in significant reduction of the serum IgE levels, which are
normally elevated in these mice following infection as a result of
sustained IL-4 production (8). When IL-4 is not produced during priming
in vitro, IL-12R ß2-chain expression is maintained on activated
CD4+ T cells, allowing IL-12 to induce Th1 cell development
independently of IFN- (13). This contention is supported by results
obtained in vivo showing that administration of exogenous IL-12 at the
onset of infection to resistant C57BL/6 mice with disruption of the
IFN- gene markedly suppressed the IL-4 mRNA expression otherwise
seen in these mice 5 days after infection (31). Furthermore, we have
recently shown that IFN-R-/- mice on a resistant
129/Sv/Ev background, although susceptible to infection as a result of
the inability of their macrophages to receive the IFN--mediated
activating signals, develop a polarized Th1 response, in contrast to
BALB/c or BALB/c IFN-R-/- mice (21). In contrast to
BALB/c IFN-R-/- mice, Sv129 IFN-R-/-
mice do not produce an early burst of IL-4 mRNA expression in their
draining lymph modes in response to L. major and maintain
IL-12R ß2-chain mRNA expression on their CD4+ T cells at
least up to 6 days after infection (P.L. and J.A.L., unpublished
results). Thus, it is likely that in the absence of IL-4, activated
CD4+ T cells do not require IFN- signaling for the
maintenance of IL-12R ß2-chain expression and IL-12 signaling in
vivo.
It has been recently shown that treatment of BALB/c mice with a soluble
protein before priming with this protein Ag in CFA diverted their
CD4+ T cell response toward the Th2 phenotype (32).
Interestingly, administration of IL-12 together with the soluble
protein Ag redirected the response to the Th1 pathway, an effect that
was attributed to the expression of IL-12R ß2-chain mRNA expression
on responding CD4+ cells (32). Even though the IFN-
dependence of this effect of IL-12 was not studied, the injection of
IFN- together with the soluble protein Ag failed to redirect the
response to the Th1 pathway (32). Results from other studies have
clearly shown that the IL-12-induced Th1 cell development from naive
precursor CD4+ T cells required IFN-, which by itself
was not sufficient to induce Th1 cell maturation (25, 33). Thus, it is
possible that treatment of BALB/c mice with soluble Ag with or without
IFN- would not lead to the production of IL-12 in amounts sufficient
for inducing Th1 cell maturation.
In summary, this study indicates that following infection of BALB/c mice with L. major, the IL-4-mediated down-regulation of IL-12R ß2-chain expression represents an important step in the development of a Th2 cell response in vivo. In-depth understanding of the factors capable of overriding this suppression of IL-12 signaling might be instrumental for the design of strategies to prevent the induction of Th2 responses detrimental to the host following infection with intracellular micro-organisms.
|
Acknowledgments |
|---|
R-/-
mice, and Drs Maurice Gately and Ueli Gubler, Department of
Inflammation/Autoimmune Diseases, Hoffmann-La Roche, Inc. (Nutley, NJ),
for providing murine rIL-12 and the IL-12R ß1- and ß2-chain cDNA,
respectively. |
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Jacques A. Louis, World Health Organization Immunology Research and Training Center, Institute of Biochemistry, University of Lausanne, Chemin des Boveresses 155, CH-1066 Epalinges, Switzerland. E-mail address:
3 On leave from the Institut Pasteur (Paris, France).
4 Abbreviation used in this paper: HPRT, hypoxanthine guanine phosphoribosyl transferase.
Received for publication May 7, 1998. Accepted for publication July 21, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
ß T-cell-receptor transgenic system. Proc. Natl. Acad. Sci. USA 89:6065. independent mechanism. J. Exp. Med. 171:115./ß interferon or the interferon receptor. J. Virol. 72:2638. receptor. Science 259:1742. receptor are susceptible to infection with Leishmania major but mount a polarized T helper cell 1-type CD4+ T cell response. J. Exp. Med. 181:961. during infection of mice with Plasmodium chabaudi chabaudi. Immunobiology 179:353.[Medline]
ß TCR transgenic mice: IL-12 substitution for macrophages to stimulate IFN- production is IFN- dependent. Int. Immunol. 5:1119.8 CD4+ T cells in BALB/c mice infected with Leishmania major instructsTh2 cell development and susceptibility to infection. Immunity 6:541.[Medline]
modulates the early development of Th1 and Th2 responses in a murine model of cutaneous leishmaniasis. J. Immunol. 147:3149.[Abstract]
-independent effects of interleukin 12 administered during acute or established infection due to Leishmania major. Proc. Natl. Acad. Sci. USA 91:12932. and IFN- in IL-12-induced T helper cell-1 development. J. Immunol. 156:1442.[Abstract]
This article has been cited by other articles:
|
|
 |
|
  S Kugathasan, L J Saubermann, L Smith, D Kou, J Itoh, D G Binion, A D Levine, R S Blumberg, and C Fiocchi Mucosal T-cell immunoregulation varies in early and late inflammatory bowel disease Gut, December 1, 2007; 56(12): 1696 - 1705. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Xu, D. Liu, I. Okwor, Y. Wang, H. Korner, S. K. P. Kung, Y.-X. Fu, and J. E. Uzonna LIGHT Is Critical for IL-12 Production by Dendritic Cells, Optimal CD4+ Th1 Cell Response, and Resistance to Leishmania major J. Immunol., November 15, 2007; 179(10): 6901 - 6909. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. G. Rhodes, J. Sawyer, A. O. Whelan, G. S. Dean, M. Coad, K. J. Ewer, A. S. Waldvogel, A. Zakher, D. J. Clifford, R. G. Hewinson, et al. Is Interleukin-4{delta}3 Splice Variant Expression in Bovine Tuberculosis a Marker of Protective Immunity? Infect. Immun., June 1, 2007; 75(6): 3006 - 3013. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Nigg, S. Zahn, D. Ruckerl, C. Holscher, T. Yoshimoto, J. M. Ehrchen, F. Wolbing, M. C. Udey, and E. von Stebut Dendritic Cell-Derived IL-12p40 Homodimer Contributes to Susceptibility in Cutaneous Leishmaniasis in BALB/c Mice J. Immunol., June 1, 2007; 178(11): 7251 - 7258. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. U. Buxbaum and P. Scott Interleukin 10- and Fc{gamma} Receptor-Deficient Mice Resolve Leishmania mexicana Lesions Infect. Immun., April 1, 2005; 73(4): 2101 - 2108. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. VANLOUBBEECK and D. E. JONES PROTECTION OF C3HEB/FEJ MICE AGAINST LEISHMANIA AMAZONENSIS CHALLENGE AFTER PREVIOUS LEISHMANIA MAJOR INFECTION Am J Trop Med Hyg, October 1, 2004; 71(4): 407 - 411. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Cameron, A. McGachy, M. Anderson, A. Paul, G. H. Coombs, J. C. Mottram, J. Alexander, and R. Plevin Inhibition of Lipopolysaccharide-Induced Macrophage IL-12 Production by Leishmania mexicana Amastigotes: The Role of Cysteine Peptidases and the NF-{kappa}B Signaling Pathway J. Immunol., September 1, 2004; 173(5): 3297 - 3304. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. F. Vanloubbeeck, A. E. Ramer, F. Jie, and D. E. Jones CD4+ Th1 Cells Induced by Dendritic Cell-Based Immunotherapy in Mice Chronically Infected with Leishmania amazonensis Do Not Promote Healing Infect. Immun., August 1, 2004; 72(8): 4455 - 4463. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. E. C. Babay, H. Louzir, C. Kebaier, S. Boubaker, K. Dellagi, and P.-A. Cazenave Inbred Strains Derived from Feral Mice Reveal New Pathogenic Mechanisms of Experimental Leishmaniasis Due to Leishmania major Infect. Immun., August 1, 2004; 72(8): 4603 - 4611. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Mattner, A. Wandersee-Steinhauser, A. Pahl, M. Rollinghoff, G. R. Majeau, P. S. Hochman, and C. Bogdan Protection against Progressive Leishmaniasis by IFN-{beta} J. Immunol., June 15, 2004; 172(12): 7574 - 7582. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Ji, J. Sun, and L. Soong Impaired Expression of Inflammatory Cytokines and Chemokines at Early Stages of Infection with Leishmania amazonensis Infect. Immun., August 1, 2003; 71(8): 4278 - 4288. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Yokota, M. Burne-Taney, L. Racusen, and H. Rabb Contrasting roles for STAT4 and STAT6 signal transduction pathways in murine renal ischemia-reperfusion injury Am J Physiol Renal Physiol, August 1, 2003; 285(2): F319 - F325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. von Stebut, J. M. Ehrchen, Y. Belkaid, S. L. Kostka, K. Molle, J. Knop, C. Sunderkotter, and M. C. Udey Interleukin 1{alpha} Promotes Th1 Differentiation and Inhibits Disease Progression in Leishmania major-susceptible BALB/c Mice J. Exp. Med., July 21, 2003; 198(2): 191 - 199. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. F. Coelho, C. A. P. Tavares, F. A. A. Carvalho, K. F. Chaves, K. N. Teixeira, R. C. Rodrigues, H. Charest, G. Matlashewski, R. T. Gazzinelli, and A. P. Fernandes Immune Responses Induced by the Leishmania (Leishmania) donovani A2 Antigen, but Not by the LACK Antigen, Are Protective against Experimental Leishmania (Leishmania) amazonensis Infection Infect. Immun., July 1, 2003; 71(7): 3988 - 3994. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Muraille, C. De Trez, M. Brait, P. De Baetselier, O. Leo, and Y. Carlier Genetically Resistant Mice Lacking MyD88-Adapter Protein Display a High Susceptibility to Leishmania major Infection Associated with a Polarized Th2 Response J. Immunol., April 15, 2003; 170(8): 4237 - 4241. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. M. Padigel and J. P. Farrell CD40-CD40 Ligand Costimulation Is Not Required for Initiation and Maintenance of a Th1-Type Response to Leishmania major Infection Infect. Immun., March 1, 2003; 71(3): 1389 - 1395. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Li, U. M. Padigel, P. Scott, and J. P. Farrell Combined Treatment with Interleukin-12 and Indomethacin Promotes Increased Resistance in BALB/c Mice with Established Leishmania major Infections Infect. Immun., October 1, 2002; 70(10): 5715 - 5720. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gorelik, S. Constant, and R. A. Flavell Mechanism of Transforming Growth Factor {beta}-induced Inhibition of T Helper Type 1 Differentiation J. Exp. Med., June 3, 2002; 195(11): 1499 - 1505. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Kuroda, T. Kito, and U. Yamashita Reduced Expression of STAT4 and IFN-{gamma} in Macrophages from BALB/c Mice J. Immunol., June 1, 2002; 168(11): 5477 - 5482. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. M. Padigel, P. J. Perrin, and J. P. Farrell The Development of a Th1-Type Response and Resistance to Leishmania major Infection in the Absence of CD40-CD40L Costimulation J. Immunol., November 15, 2001; 167(10): 5874 - 5879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Helmby, K. Takeda, S. Akira, and R. K. Grencis Interleukin (Il)-18 Promotes the Development of Chronic Gastrointestinal Helminth Infection by Downregulating IL-13 J. Exp. Med., August 6, 2001; 194(3): 355 - 364. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Finotto, G. T. De Sanctis, H. A. Lehr, U. Herz, M. Buerke, M. Schipp, B. Bartsch, R. Atreya, E. Schmitt, P. R. Galle, et al. Treatment of Allergic Airway Inflammation and Hyperresponsiveness by Antisense-Induced Local Blockade of Gata-3 Expression J. Exp. Med., June 4, 2001; 193(11): 1247 - 1260. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Nishikomori, S. Gurunathan, K. Nishikomori, and W. Strober BALB/c Mice Bearing a Transgenic IL-12 Receptor {{beta}}2 Gene Exhibit a Nonhealing Phenotype to Leishmania major Infection Despite Intact IL-12 Signaling J. Immunol., June 1, 2001; 166(11): 6776 - 6783. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kumar and R. L. Tarleton Antigen-Specific Th1 But Not Th2 Cells Provide Protection from Lethal Trypanosoma cruzi Infection in Mice J. Immunol., April 1, 2001; 166(7): 4596 - 4603. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. I. Araujo, S. K. Bliss, Y. Suzuki, A. Alcaraz, E. Y. Denkers, and E. J. Pearce Interleukin-12 Promotes Pathologic Liver Changes and Death in Mice Coinfected with Schistosoma mansoni and Toxoplasma gondii Infect. Immun., March 1, 2001; 69(3): 1454 - 1462. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Schilling and N. Glaichenhaus T Cells That React to the Immunodominant Leishmania major LACK Antigen Prevent Early Dissemination of the Parasite in Susceptible BALB/c Mice Infect. Immun., February 1, 2001; 69(2): 1212 - 1214. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhu, H. Huang, L. Guo, T. Stonehouse, C. J. Watson, J. Hu-Li, and W. E. Paul Transient Inhibition of Interleukin 4 Signaling by T Cell Receptor Ligation J. Exp. Med., October 16, 2000; 192(8): 1125 - 1134. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Elhofy, I. Marriott, and K. L. Bost Salmonella Infection Does Not Increase Expression and Activity of the High Affinity IL-12 Receptor J. Immunol., September 15, 2000; 165(6): 3324 - 3332. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Vos, C. M. Snapper, and J. J. Mond Th1 versus Th2 cytokine profile determines the modulation of in vitro T cell-independent type 2 responses by IL-4 Int. Immunol., September 1, 2000; 12(9): 1337 - 1345. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Tacchini-Cottier, C. Zweifel, Y. Belkaid, C. Mukankundiye, M. Vasei, P. Launois, G. Milon, and J. A. Louis An Immunomodulatory Function for Neutrophils During the Induction of a CD4+ Th2 Response in BALB/c Mice Infected with Leishmania major J. Immunol., September 1, 2000; 165(5): 2628 - 2636. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Sato, S. K. Ahuja, M. Quinones, V. Kostecki, R. L. Reddick, P. C. Melby, W. A. Kuziel, and S. S. Ahuja Cc Chemokine Receptor (Ccr)2 Is Required for Langerhans Cell Migration and Localization of T Helper Cell Type 1 (Th1)-Inducing Dendritic Cells: Absence of Ccr2 Shifts the Leishmania major-Resistant Phenotype to a Susceptible State Dominated by Th2 Cytokines, B Cell Outgrowth, and Sustained Neutrophilic Inflammation J. Exp. Med., July 17, 2000; 192(2): 205 - 218. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. E. Jones, L. U. Buxbaum, and P. Scott IL-4-Independent Inhibition of IL-12 Responsiveness During Leishmania amazonensis Infection J. Immunol., July 1, 2000; 165(1): 364 - 372. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Himmelrich, P. Launois, I. Maillard, T. Biedermann, F. Tacchini-Cottier, R. M. Locksley, M. Rocken, and J. A. Louis In BALB/c Mice, IL-4 Production During the Initial Phase of Infection with Leishmania major Is Necessary and Sufficient to Instruct Th2 Cell Development Resulting in Progressive Disease J. Immunol., May 1, 2000; 164(9): 4819 - 4825. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Mohrs, C. Holscher, and F. Brombacher Interleukin-4 Receptor Alpha-Deficient BALB/c Mice Show an Unimpaired T Helper 2 Polarization in Response to Leishmania major Infection Infect. Immun., April 1, 2000; 68(4): 1773 - 1780. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Yin, D.-H. Zhang, T. Welte, G. Bahtiyar, S. Jung, L. Liu, X.-Y. Fu, A. Ray, and J. Craft Dominance of IL-12 Over IL-4 in {gamma}{delta} T Cell Differentiation Leads to Default Production of IFN-{gamma}: Failure to Down-Regulate IL-12 Receptor {beta}2-Chain Expression J. Immunol., March 15, 2000; 164(6): 3056 - 3064. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Nishikomori, R. O. Ehrhardt, and W. Strober T Helper Type 2 Cell Differentiation Occurs in the Presence of Interleukin 12 Receptor {beta}2 Chain Expression and Signaling J. Exp. Med., March 6, 2000; 191(5): 847 - 858. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Kuroda, T. Sugiura, K. Zeki, Y. Yoshida, and U. Yamashita Sensitivity Difference to the Suppressive Effect of Prostaglandin E2 Among Mouse Strains: A Possible Mechanism to Polarize Th2 Type Response in BALB/c Mice J. Immunol., March 1, 2000; 164(5): 2386 - 2395. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. P. Heinzel and R. A. Maier Jr. Interleukin-4-Independent Acceleration of Cutaneous Leishmaniasis in Susceptible BALB/c Mice following Treatment with Anti-CTLA4 Antibody Infect. Immun., December 1, 1999; 67(12): 6454 - 6460. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. K. Thibodeaux, S. E. Hunter, K. E. Waldburger, J. L. Bliss, W. L. Trepicchio, J. P. Sypek, K. Dunussi-Joannopoulos, S. J. Goldman, and J. P. Leonard Autocrine Regulation of IL-12 Receptor Expression Is Independent of Secondary IFN-{gamma} Secretion and not Restricted to T and NK Cells J. Immunol., November 15, 1999; 163(10): 5257 - 5264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. R. Schopf, J. L. Bliss, L. M. Lavigne, C. L. Chung, S. F. Wolf, and J. P. Sypek Interleukin-12 Is Capable of Generating an Antigen-Specific Th1-Type Response in the Presence of an Ongoing Infection-Driven Th2-Type Response Infect. Immun., May 1, 1999; 67(5): 2166 - 2171. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.G. Begley and N.A. Nicola Resolving Conflicting Signals: Cross Inhibition of Cytokine Signaling Pathways Blood, March 1, 1999; 93(5): 1443 - 1447. [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
