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-Inducing Factor Prevent Experimental Autoimmune Encephalomyelitis1
*
Department of Immunology and
Rappaport Family Institute for Research in the Medical Sciences and Bruce Rappaport Faculty of Medicine, Haifa, Israel
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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-inducing factor (IGIF) from the brain of rats with developing
experimental autoimmune encephalomyelitis (EAE), a T cell-mediated
autoimmune disease of the central nervous system that serves as a model
for multiple sclerosis. IGIF was highly transcribed in the brain at the
onset and during the course of active EAE. PCR products encoding rat
IGIF were used to generate the recombinant protein that was used to
induce anti-IGIF neutralizing Abs. These Abs significantly reduced
the production of IFN- by primed T cells proliferating in response
to their target myelin basic protein epitope and by Con A-activated T
cells from naive donors. When administered to rats during the
development of either active or transferred EAE, these Abs
significantly blocked the development of disease. Splenic T cells from
protected rats were cultured with the encephalitogenic myelin basic
protein epitope and evaluated for production of IL-4 and IFN-. These
cells, which proliferated, exhibited a profound increase in IL-4
production that was accompanied by a significant decrease in IFN-
and TNF-
production. Thus, we suggest that perturbation of the
Th1/Th2 balance toward Th2 cells is the mechanism underlying EAE
blockade by anti-IGIF immunotherapy. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and TNF- and, to a
much lesser extent, IL-4 and IL-10; Th2 cells that produce IL-4, IL-10,
and IL-13 and, to a much lesser extent, IFN- and TNF- (1, 2, 3, 4, 5, 6, 7, 8, 9, 10);
and the newly defined Th3 cells that produce significant amounts of
TGF-ß and have been associated with oral tolerance (11). Th1 cells,
which are selected in response to various autoantigens, transfer T
cell-mediated autoimmune diseases; IL-4-secreting Th2 cells, which are
selected in response to these same Ags, either inhibit the inflammatory
process or exert no profound effect (5, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24). High levels of
IFN- and low levels of IL-4 positively select for Th1 cells, whereas
low levels of IFN- together with high levels of IL-4 mediate Th2
selection (1, 2, 3, 4, 5, 6).
IFN--inducing factor
(IGIF)3 (IL-18) is a recently
described cytokine (25) that shares structural features with the IL-1
family of proteins (26). The activation of IGIF is mediated by
IL-1ß-converting enzyme (27, 28). Like IL-12, IGIF is a potent
inducer of the production of IFN- by Th1 and NK cells and acts on
Th1 cells together with IL-12 in a synergistic manner (25, 29, 30, 31, 32).
Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated autoimmune disease of the central nervous system that serves as a model for human multiple sclerosis (33, 34). Ag-specific T cells are thought to play a pivotal role in the manifestation of both diseases (35, 36, 37).
The role of Th1 cells in the manifestation of EAE has been widely
studied. Th1 but not Th2 cells transfer the disease to normal naive
recipients (18). Shifting the Th1/Th2 balance toward Th2 cells by the
in vivo administration of IL-4 (12), by Abs to B7-1 (14), by soluble
peptide therapy (38), or by the administration of neutralizing Abs to
IL-12 (15) markedly suppressed EAE. It has been shown recently that
IGIF is a more potent inducer of IFN--producing Th1 cells than is
IL-12 and consequently plays an important role in Th1 responses (25).
However, the possible role of anti-IGIF immunotherapy in the
regulation of T cell-mediated autoimmunity has never been evaluated.
The current study demonstrates, for the first time, that neutralizing Abs to IGIF ameliorate EAE by shifting the Th1/Th2 balance toward Ag-specific Th2 cells.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Female Lewis rats (6 wk old) were purchased from Harlan (Jerusalem, Israel) and maintained under specific pathogen-free conditions in our animal facility.
Peptide Ags
Myelin basic protein (MBP) p68–86 (Y G S L P Q K S Q R S Q D E N P V) was synthesized on a Milligen 9050 peptide synthesizer (Burlington, MA) by standard 9-fluorenylmethoxycarbonyl chemistry. Peptides were purified by HPLC. Structure was confirmed by amino acid analysis and mass spectroscopy. Only peptides that were >95% pure were used in our study.
Immunizations and induction of active disease
Rats were immunized s.c. in the hind footpads with 0.1 ml of MBP epitope 68–86 (p68–86) dissolved in PBS (1.5 mg/ml) and emulsified with an equal volume of CFA (IFA supplemented with 4 mg/ml heat-killed Mycobacterium tuberculosis H37Ra in oil (Difco, Detroit, MI)). Rats were then monitored daily for clinical signs by an observer who was blind to the treatment protocol. EAE was scored as follows: 0, clinically normal; 1, flaccid tail; 2, hind limb paralysis; and 3, front and hind limb paralysis.
Induction of transferred EAE
EAE was induced by immunizing Lewis rats i.p. with 108 activated spleen cells from EAE donors. Cells were obtained as follows: At 9 days after the induction of active EAE, splenic cells were cultured (12 x 106/ml) at 37°C in humidified air containing 7.5% CO2 for 2 days in stimulation medium that included DMEM (Life Technologies, Gaithersburg, MD) supplemented with 2-ME (5 x 10-5 M), L-glutamine (2 mM), sodium pyruvate (1 mM), penicillin (100 µ/ml), streptomycin (100 µg/ml), 1% syngeneic serum, and 20–30 µg/ml of the immunizing epitope. Next, cells were separated on a Ficoll gradient (Sigma, St. Louis, MO), resuspended in PBS, and injected into naive recipients.
Ag-specific T cell proliferation assays
Lewis rats were immunized with MBP p68–86/CFA as described above. After 9–10 days, spleen cells were suspended in stimulation medium and cultured in U-shaped 96-well microculture plates (2 x 105 cells/well) for 72 h at 37°C in humidified air containing 7.5% CO2. Each well was pulsed with 2 µCi of [3H]thymidine (specific activity of 10 Ci/mmol) for the final 6 h. The cultures were then harvested on fiberglass filters, and the proliferative response was expressed as cpm ± SD or as stimulation index (SI) (mean cpm of test cultures divided by mean cpm of control cultures).
RT-PCR analysis
RT-PCR analysis, verified by Southern blotting, was used on
brain samples according to the protocol we described elsewhere with
some modifications (39). Rats were euthanized by CO2
narcosis. Brain samples containing mainly the midbrain and brain stem
were obtained after perfusion of the rat with 160–180 ml of ice-cold
PBS injected into the left ventricle following an incision in the right
atrium. Each sample was homogenized. Total RNA was extracted using the
Tri-Zol procedure (Life Technologies) according to the manufacturer’s
protocol. mRNA was then isolated using an mRNA isolation kit (No.
1741985, Boheringer Mannheim, Mannheim, Germany) and was reverse
transcribed into first-strand cDNA as we have described in detail
elsewhere (39). First-strand cDNA was then subjected to 35 cycles of
PCR amplification using specific oligonucleotide primers to rat IGIF
and IFN- that we designed based on the published sequence of each
cytokine (National Center for Biotechnology Information
accession numbers for rat IGIF U77777 and rat IFN- M29315) as
follows: rat IGIF sense, 5'-ATGGCTGCCATGTCAGAAGAAG-3'; rat IGIF
antisense, 5'-CTAACTTTGATGTAAGTTAGTAAGA-3'; rat IFN- sense,
5'-TACTGCCAAGGCACACTCATTGAA-3'; rat IFN- antisense,
5'-CGCTTCCTTAGGCTAGATTCTGG-3'; rat ß-actin sense,
5'-CATCGTGGGCCGCTCTAGGCA-3' (39); and rat ß-actin antisense,
5'-CCGGCCAGCCAAGTCCAGACG-3' (39).
Experimental conditions were calibrated so that RT-PCR amplifications
fell on the linear part of the titration curve. The cycle profile was
denaturation at 95°C for 60 s, annealing at 55°C for 60
s, and elongation at 72°C for 60 s. Amplified products were
subjected to electrophoresis, transferred to a nylon membranes
(MagnaGraph nylon transfer membrane, Micron Separations, Westborough,
MA), fixed with UV light (120 mjoules), and hybridized with
106 cpm/ml of -32P-labeled DNA
fragments encoding the full-length PCR product of IGIF and ß-actin
(random priming: Amersham, Arlington Heights, IL). PCR products were
used as probes only after each PCR product was cloned; the sequence of
each PCR product was verified as described below. Southern blot
images were objectively assessed using a FujiFilm Thermal System
FTI-500 (FujiFilm, Tokyo, Japan).
Cloning and sequencing of PCR products
Each of the amplified PCR products described above was cloned into a pUC57/T vector (T-cloning kit No. K1212, MBI Fermentas, Vilnius, Lithuania) and transformed to Escherichia coli according to the manufacturer’s protocol. Each clone was then sequenced (Sequenase version 2, United States Biochemical, Cleveland, Ohio) according to the manufacturer’s protocol.
Production and purification of recombinant proteins
After sequence verification, PCR products were recloned into a pQE expression vector (pQE-30, pQE-31, or pQE-32 according the correct open reading frame) and expressed in E. coli (Qiagen, Hilden, Germany) and purified by an Ni-nitrilotriacetic acid super flow affinity purification of 6xHis proteins (Qiagen). Each recombinant protein sequence has been verified (N terminus) by our sequencing services unit.
Production and purification of rabbit anti-rat IGIF IgG
Rabbit anti-rat IGIF Abs were generated as described previously (40); the IgG fraction was purified using a HiTrap protein G kit (No. 17-040-01, Pharmacia, Piscataway, NJ). Ab titer was determined by direct ELISA: ELISA plates (Nunc, Roskilde, Denmark) were coated with rat rIGIF (50 ng/well). Rabbit anti-rat IGIF (IgG fraction) was added in serial dilutions from 28 to 230. Goat anti-rabbit IgG alkaline phosphatase (AP)-conjugated Abs (Sigma) were used as a labeled Ab. p-nitrophenyl phosphate (Sigma) was used as a soluble AP substrate. The results of triplicate cultures were calculated as log2 Ab titer ± SE. Our purified anti-rat IGIF IgG titer was 18 ± 0.4.
Cytokine determination
Spleen cells from EAE donors were stimulated in vitro
(107 cells/ml) in 24-well plates (Nunc) with 100 µM of
p68–86. Spleen cells from naive donors were cultured (107
cells/ml, 24-well plates) with 2 µg/ml Con A (Sigma). After 72 h
of stimulation, supernatants were assayed using semiELISA kits that
included Ab pairs and recombinant rat cytokines as follows: for
IFN-, rabbit anti-rat IFN- polyclonal Ab (CY-048,
Innogenetics, Zwijnaarde, Belgium) was used as a capture Ab,
biotinylated mouse anti-rat mAb (CY-106 clone BD-1, Innogenetics)
was used as a detection Ab, and AP-streptavidin (catalog No. 43-4322,
Zymed, San Francisco, CA) with rat rIFN- was used as a
standard (catalog No. 3281SA, Life Technologies); for TNF-, a
commercial semiELISA kit was used for the detection of rat TNF-
(catalog No. 80-3807-00, Genzyme, Cambridge, MA); for IL-4, mouse
anti-rat IL-4 mAb (24050D OX-81, PharMingen, San Diego, CA) was
used as a capture Ab and rabbit anti-rat IL-4 biotin-conjugated
polyclonal Ab (2411-2D, PharMingen) as second Ab. Rat rIL-4 that
was purchased from R&D Systems (Minneapolis, MN) (504-RL) was
used as a standard.
Statistical analysis
The significance of differences was examined using the Student
t test. The Mann-Whitney sum of ranks test was used to
evaluate the significance of differences in the mean of the maximal
clinical score (see Fig. 3
). A p value of <0.05 was
considered significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Midbrain-brain stem samples were obtained from rats with
developing transferred EAE (Fig. 1A) before adoptive transfer
of disease (day 0), before the onset of disease (day 3), at the day of
onset (day 5), at the peak of disease (day 7), following recovery (day
10), and at 10 days postrecovery (day 20). From each timepoint, samples
from six different brains were subjected to RT-PCR analysis using
specific oligonucleotide primers that we constructed to IGIF and
IFN-. Each amplification was calibrated to ß-actin and verified by
Southern blotting analysis, which enabled semiquantitative
analysis of the dynamics of mRNA transcription of IGIF and IFN- at
the site of inflammation. Fig. 1, C and E show
representative results from each timepoint of the experiment. A
substantial increase in the transcription of both IGIF and IFN- mRNA
in EAE brains was observed at the peak of disease (day 7). The
augmented transcription of IFN- mRNA reverted to background during
recovery. Unexpectedly, a notable transcription of IGIF mRNA could be
observed even at 10 days postrecovery (Fig. 1C).
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mRNA in EAE brains was observed
at the peak of disease (day 13). The augmented transcription of
IFN-, but not of IGIF mRNA, regressed to background during recovery
(Fig. 1
, D and F). A substantial increase in the
level of IGIF transcription at the site of inflammation in the central
nervous system during the course of disease may suggest involvement in
disease regulation. To evaluate this point, the role of IGIF in
the regulation of EAE was investigated.
Rat rIGIF and its neutralizing Abs affect IFN- production by
activated T cells from naive donors more significantly than by
Ag-specific-primed T cells
PCR products encoding rat IGIF were used to generate the
recombinant protein that was used to produce anti-IGIF neutralizing
Abs. These Abs significantly reduced the production of IFN- in the
primed T cells proliferating in response to their specific MBP epitope
(Fig. 2C, 3.2 ± 0.25 vs
1.8 ± 0.11 ng/ml with backgrounds of 0.2 ± 0.1 and
0.25 ± 0.1; p < 0.01) and entirely blocked
IFN- production in Con A-activated T cells from naive donors (Fig. 2A, 5.1 ± 0.4 vs 0.42 ± 0.1 ng/ml with
backgrounds of 0.4 ± 0.1 and 0.36; p < 0.001).
Control IgG from normal rabbit serum did not exert a notable effect on
IFN- production by either Con A-activated naive spleen cells or MBP
p68–86-primed spleen cells (data not shown). Rat rIGIF elicited
IFN- production in Con A-activated splenic T cells from naive donors
(Fig. 2B, 15.8 ± 0.8 ng/ml vs 5.1 ± 0.3 with
backgrounds of 0.3 ± 0.1 and 0.4 ± 0.15; p
< 0.001) and significantly although again less profoundly influenced
the response of primed spleen T cells to their target MBP Ag (Fig. 2D, 4.97 ± 0.15 ng/ml vs 3.2 ± 0.25 with
backgrounds of 0.3 ± 0.15 and 0.25 ± 0.1; p
< 0.001). Thus, both the inhibitory effect of IGIF-neutralizing Abs
and the augmentation by IGIF of IFN- production are more profound on
activated T cells from a naive donor than on primed T cells responding
to their target epitope. It has recently been suggested that IGIF
primarily affects IFN- production by Th1 rather than Th2 cells (29).
It is possible that immunization with p68–86/CFA induces a substantial
selection of Ag-specific Th2 cells, albeit not enough to inhibit the
subsequent development of a Th1-mediated autoimmune disease.
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Neutralizing Abs to rat rIGIF block the development of both active and transferred EAE
The role of anti-IGIF Abs in the regulation of T cell-mediated
autoimmune diseases has never been explored. We have evaluated the
competence of the anti-IGIF neutralizing Abs with regard to the
inhibition of active (Fig. 3A) and transferred (Fig. 3B) EAE. Lewis rats were immunized with p68–86/CFA to
develop active EAE. Just before the onset of disease (days 8 and 10)
and at the onset of disease (day 11), these rats were injected with
either rabbit anti-rat IGIF (IgG fraction), IgG fraction purified
from nonimmunized rabbits (control IgG), or PBS and monitored for
clinical signs of EAE. Control PBS-treated rats and rats treated with
control IgG all (six of six rats in each group) developed severe EAE
(mean maximal clinical scores were 3.3 ± 0.43 and 2.66 ±
0.26, respectively). In contrast, rats treated with anti-IGIF Abs
developed a markedly reduced disease (Fig. 3A, incidence was
in five of six rats, mean maximal clinical score of 1.2 ± 0.2;
p < 0.01). We have further evaluated the competence of
anti-IGIF Abs in the inhibition of transferred EAE (Fig. 3B). At 3, 5, and 7 days after adoptive transfer of disease,
rats were injected as described above and monitored for clinical signs
of EAE. Although control PBS-treated rats and rats treated with control
IgG all (six of six rats in each group) developed EAE (mean maximal
clinical score was 1 ± 0 in each group), the rats that were
administered anti-IGIF Abs were highly protected (Fig. 3B, incidence was in one of six rats, mean maximal clinical
score was 0.2 ± 0.1; p < 0.01). Thus,
immunotherapy with anti-IGIF may serve as a powerful tool to block
the development of actively induced or transferred EAE.
Alteration of IFN- and IL-4 production in EAE rats injected with
anti-IGIF neutralizing Abs suggests that perturbation of the
Th2/Th1 balance contributes to disease blockade
The possible involvement of a Th2/Th1 switch in EAE inhibition by
anti-IGIF immunotherapy has been evaluated (Fig. 4). Lewis rats were immunized with
p68–86/CFA to develop active EAE. After 5 and 7 days, these rats were
injected with either PBS, control rabbit IgG, or rabbit anti-rat
IGIF (IgG fraction). At 2 days after the last treatment, splenic T
cells were cultured with MBP p68–86 in stimulation medium that was
(Fig. 4, C and D) or was not (Fig. 4, A and B) supplemented with rat rIL-4. In spleen
cells cultured from MBP 68–86-primed donors, IFN- was produced only
when the priming Ag was added to the culture (Fig. 4A,
0.3 ± 0.1 ng/ml without the addition of MBP 68–86 vs 13.5
± 0.7 in cells proliferating to p68–86). The addition of rIL-4 led to
a significant decrease in IFN- that was still dependent upon
antigenic stimulation (Fig. 4, A and C, 0.19
± 0.08 ng/ml without the addition of MBP 68–86 vs 2.37 ± 0.8 in
cells proliferating to p68–86; a 12-fold increase). Spleen T cells
from anti-IGIF-treated rats produced markedly reduced levels of
IFN- in response to antigenic stimulation in cultures that were or
were not supplemented with IL-4 (Fig. 4A, 4.7 ± 0.4
ng/ml in spleen cells from anti-IGIF-treated rats vs 9.7 ±
0.8 in spleen cells from rats treated with normal rabbit IgG and
13.5 ± 0.7 in PBS-treated rats with backgrounds of 0.4, 0.8, and
0.7; p < 0.001 when comparing anti-IGIF treatment
with each control group). However, IL-4 production markedly increased
in splenic T cells from anti-IGIF-treated rats regardless of in
vitro stimulation (Fig. 4B, 62.3 ± 4.2 pg/ml in spleen
cells from anti-IGIF-treated rats vs 15.3 ± 0.4 in spleen
cells from rats treated with normal rabbit IgG and 15.6 ± 0.6 in
PBS-treated rats; p < 0.001 when comparing
anti-IGIF treatment with each control group) unless cultures were
supplemented with IL-4 (Fig. 4D, 1860 ± 120 pg/ml in
spleen cells from anti-IGIF-treated rats vs 570 ± 30 in
spleen cells from rats treated with normal rabbit IgG and 450 ±
35 in PBS-treated rats with backgrounds of 85, 42, and 34;
p < 0.0001 when comparing anti-IGIF treatment with
each control group). The addition of IL-4 to cultured spleen T cells
(Fig. 4, C–D) did not exhibit a notable effect on the
Ag-specific proliferative response of these cells (data not
shown).
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production was then evaluated in spleen cells from the above
groups. The above spleen cells from anti-IGIF-treated rats produced
markedly reduced levels of TNF- in response to antigenic stimulation
(Fig. 5, 850 ± 45 pg/ml in spleen
cells from anti-IGIF-treated rats vs 1975 ± 80 in spleen
cells from rats treated with normal rabbit IgG and 2100 ± 110 in
PBS-treated rats with backgrounds of 230, 210, and 270, reciprocally;
p < 0.001 when comparing anti-IGIF treatment with
each control group). Thus, perturbation of the Th1/Th2 balance in
anti-IGIF-treated rats is associated with a marked reduction in
TNF- production.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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positively select for
TNF--secreting Th1 cells (6). A previous study showed that the
administration of anti-IL-12 neutralizing Abs blocks EAE while
inducing a marked reduction of both IFN- and TNF- production
(15). IFN- and TNF- in combination subsequently exhibit a
synergistic effect on the enhancement of expression of adhesion
molecules on endothelial cells (44) and on the elicitation of the
inflammatory process, which can be reversed by either antiadhesion
molecule immunotherapy (45, 46) or by blocking TNF- (44, 47, 48, 49, 50). We have demonstrated previously that the EAE
resistance acquired by soluble Ag therapy can be reversed by
anti-IL-4 neutralizing Abs (38). This finding further demonstrated
the pivotal role of the Th1/Th2 balance in the regulation of T
cell-mediated autoimmunity (38).
IGIF is a recently described cytokine (25) that shares structural
features with the IL-1 family of proteins (26). The activation of IGIF
is mediated by IL-1ß-converting enzyme (27, 28). Like IL-12, IGIF is
a potent inducer of IFN- from Th1 and NK cells and acts on Th1 cells
together with IL-12 in a synergistic manner (25, 29, 30, 31, 32). IGIF actually
has more potent IFN--inducing capabilities than IL-12 and apparently
uses a distinct signal transduction pathway for its elicitation (25, 31, 32, 51). Little is known about the role of IGIF in T cell-mediated
autoimmune disease. A recent study used RT-PCR to demonstrate that the
active stage of autoimmune diabetes in nonobese diabetic mice
is associated with the expression of IGIF (52). Our present study
demonstrates for the first time an elevated expression of IGIF (Fig. 1)
at the time when the secondary influx of autoimmune cells is apparent
at the site of inflammation in the EAE brain (38, 39, 46, 53);
our study also used neutralizing Abs (Fig. 2), which we generated
against the IGIF cloned from this site of inflammation, to block the
disease (Fig. 3) by altering the in vivo Th1/Th2 balance in favor of
Th2 selection. This alteration included a marked reduction in the
production of IFN- and, most importantly, TNF-, a proinflammatory
cytokine that plays a critical role in T cell-mediated autoimmunity
(44, 47, 48, 49, 50).
An interesting observation of the current study is that both the
inhibitory effect of IGIF-neutralizing Abs and the augmentation by IGIF
of IFN- production are more profound on activated T cells from a
naive donor than on primed T cells responding to their target epitope
(Fig. 2). The idea of adding rat rIL-4 to cultured T cells (Fig. 4)
came from a recent study of Lederer et al., who used the same strategy
to demonstrate that IL-4 acts on proliferating Th2 cells in an
autocrinic manner (4). It is not clear which observation contributes
more to the understanding of the in vivo changes that occur following
the administration of IGIF-neutralizing Abs: the elevated levels of
IL-4 produced by spleen T cells, even though the encephalitogenic
determinant was not added to the culture medium (Fig. 4B),
or the acceleration in IL-4 production in cultured spleen cells in
response to the encephalitogenic determinant in the presence of IL-4.
Nevertheless, both observations taken together suggest a perturbation
of the Th1/Th2 balance toward IL-4-secreting T cells in EAE rats that
were administered IGIF-neutralizing Abs.
The direct role of IFN- in EAE is enigmatic. Grewal et al. used CD40
ligand-deficient mice that carry a transgenic TCR specific for
MBP to demonstrate that EAE induction is IFN--dependent (54).
Alternatively, not only were mice lacking IFN- susceptible to the
induction of active EAE (55) but Abs to IFN- were also found to be
capable of enhancing this disease (56, 57). A recent study demonstrated
that IL-12 is directly involved in the generation of autoreactive Th1
cells that induce EAE, both in the presence and absence of IFN-
(58). However, it could well be that alteration the Th1/Th2 balance
toward IL-4-secreting Th2 cells confers EAE resistance not because it
leads to a reduced production of IFN- (Fig. 4), but rather because
it results in a reduced production of TNF- (Fig. 5) accompanied by a
marked increase in IL-4 production (Fig. 4).
It has been suggested recently that IGIF primarily affects IFN-
production by Th1 but not Th2 cells (29). It is possible that
immunization with p68–86/CFA induces a substantial selection of
Ag-specific Th2 cells, albeit not enough to inhibit the subsequent
development of a Th1-mediated autoimmune disease. Hence, as shown in
our study, the in vivo administration of anti-IGIF neutralizing Abs
notably shifts the Th1/Th2 balance in Ag-specific proliferating T cells
toward a Th2 response (Fig. 4), thus providing the immune system with a
mechanism by which it restrains its harmful activity.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Nathan Karin, Rappaport Family Institute for Research in the Medical Sciences and Bruce Rappaport Faculty of Medicine, Technion, P.O.B. 9697, Haifa 31096, Israel. E-mail address:
3 Abbreviations used in this paper: IGIF, IFN--inducing factor; EAE, experimental autoimmune encephalomyelitis; MBP, myelin basic protein; SI, stimulation index; AP, alkaline phosphatase.
Received for publication April 24, 1998. Accepted for publication July 1, 1998.
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S. Schif-Zuck, J. Westermann, N. Netzer, Y. Zohar, M. Meiron, G. Wildbaum, and N. Karin Targeted Overexpression of IL-18 Binding Protein at the Central Nervous System Overrides Flexibility in Functional Polarization of Antigen-Specific Th2 Cells J. Immunol., April 1, 2005; 174(7): 4307 - 4315. [Abstract] [Full Text] [PDF]
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R. Goldberg, Y. Zohar, G. Wildbaum, Y. Geron, G. Maor, and N. Karin Suppression of Ongoing Experimental Autoimmune Encephalomyelitis by Neutralizing the Function of the p28 Subunit of IL-27 J. Immunol., November 15, 2004; 173(10): 6465 - 6471. [Abstract] [Full Text] [PDF]
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 P. Bossu, D. Neumann, E. Del Giudice, A. Ciaramella, I. Gloaguen, G. Fantuzzi, C. A. Dinarello, E. Di Carlo, P. Musiani, P. L. Meroni, et al. IL-18 cDNA vaccination protects mice from spontaneous lupus-like autoimmune disease PNAS, November 25, 2003; 100(24): 14181 - 14186. [Abstract] [Full Text] [PDF]
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G.-X. Zhang, S. Yu, B. Gran, J. Li, I. Siglienti, X. Chen, D. Calida, E. Ventura, M. Kamoun, and A. Rostami Role of IL-12 Receptor {beta}1 in Regulation of T Cell Response by APC in Experimental Autoimmune Encephalomyelitis J. Immunol., November 1, 2003; 171(9): 4485 - 4492. [Abstract] [Full Text] [PDF]
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C. Wu, P. Sakorafas, R. Miller, D. McCarthy, S. Scesney, R. Dixon, and T. Ghayur IL-18 Receptor {beta}-Induced Changes in the Presentation of IL-18 Binding Sites Affect Ligand Binding and Signal Transduction J. Immunol., June 1, 2003; 170(11): 5571 - 5577. [Abstract] [Full Text] [PDF]
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A. Ito, A. Matejuk, C. Hopke, H. Drought, J. Dwyer, A. Zamora, S. Subramanian, A. A. Vandenbark, and H. Offner Transfer of Severe Experimental Autoimmune Encephalomyelitis by IL-12- and IL-18-Potentiated T Cells Is Estrogen Sensitive J. Immunol., May 1, 2003; 170(9): 4802 - 4809. [Abstract] [Full Text] [PDF]
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G.-X. Zhang, B. Gran, S. Yu, J. Li, I. Siglienti, X. Chen, M. Kamoun, and A. Rostami Induction of Experimental Autoimmune Encephalomyelitis in IL-12 Receptor-{beta}2-Deficient Mice: IL-12 Responsiveness Is Not Required in the Pathogenesis of Inflammatory Demyelination in the Central Nervous System J. Immunol., February 15, 2003; 170(4): 2153 - 2160. [Abstract] [Full Text] [PDF]
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 J. A. Gracie, S. E. Robertson, and I. B. McInnes Interleukin-18 J. Leukoc. Biol., February 1, 2003; 73(2): 213 - 224. [Abstract] [Full Text] [PDF]
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D. Sewell, Z. Qing, E. Reinke, D. Elliot, J. Weinstock, M. Sandor, and Z. Fabry Immunomodulation of experimental autoimmune encephalomyelitis by helminth ova immunization Int. Immunol., January 1, 2003; 15(1): 59 - 69. [Abstract] [Full Text] [PDF]
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M. Abraham, S. Shapiro, N. Lahat, and A. Miller The role of IL-18 and IL-12 in the modulation of matrix metalloproteinases and their tissue inhibitors in monocytic cells Int. Immunol., December 1, 2002; 14(12): 1449 - 1457. [Abstract] [Full Text] [PDF]
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 M. Hedtjarn, A.-L. Leverin, K. Eriksson, K. Blomgren, C. Mallard, and H. Hagberg Interleukin-18 Involvement in Hypoxic-Ischemic Brain Injury J. Neurosci., July 15, 2002; 22(14): 5910 - 5919. [Abstract] [Full Text] [PDF]
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G. Wildbaum, N. Netzer, and N. Karin Plasmid DNA Encoding IFN-{gamma}-Inducible Protein 10 Redirects Antigen-Specific T Cell Polarization and Suppresses Experimental Autoimmune Encephalomyelitis J. Immunol., June 1, 2002; 168(11): 5885 - 5892. [Abstract] [Full Text] [PDF]
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B. Wang, C. Feliciani, B. G. Howell, I. Freed, Q. Cai, H. Watanabe, and D. N. Sauder Contribution of Langerhans Cell-Derived IL-18 to Contact Hypersensitivity J. Immunol., April 1, 2002; 168(7): 3303 - 3308. [Abstract] [Full Text] [PDF]
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J. A. Jordan, R.-F. Guo, E. C. Yun, V. Sarma, R. L. Warner, L. D. Crouch, G. Senaldi, T. R. Ulich, and P. A. Ward Role of IL-18 in Acute Lung Inflammation J. Immunol., December 15, 2001; 167(12): 7060 - 7068. [Abstract] [Full Text] [PDF]
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 B. Anlar, F. Soylemezoglu, S. Aysun, G. Kose, D. Belen, and K. Yalaz Tissue Inflammatory Response in Subacute Sclerosing Panencephalitis (SSPE) J Child Neurol, December 1, 2001; 16(12): 895 - 900. [Abstract] [PDF]
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 S.-H. IM, D. BARCHAN, P. K. MAITI, L. RAVEH, M. C. SOUROUJON, and S. FUCHS Suppression of experimental myasthenia gravis, a B cell-mediated autoimmune disease, by blockade of IL-18 FASEB J, October 1, 2001; 15(12): 2140 - 2148. [Abstract] [Full Text] [PDF]
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T. Ogura, H. Ueda, K. Hosohara, R. Tsuji, Y. Nagata, S.-i. Kashiwamura, and H. Okamura Interleukin-18 stimulates hematopoietic cytokine and growth factor formation and augments circulating granulocytes in mice Blood, October 1, 2001; 98(7): 2101 - 2107. [Abstract] [Full Text] [PDF]
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 M. Neighbors, X. Xu, F. J. Barrat, S. R. Ruuls, T. Churakova, R. Debets, J. F. Bazan, R. A. Kastelein, J. S. Abrams, and A. O'Garra A Critical Role for Interleukin 18 in Primary and Memory Effector Responses to Listeria monocytogenes That Extends Beyond Its Effects on Interferon {gamma} Production J. Exp. Med., August 6, 2001; 194(3): 343 - 354. [Abstract] [Full Text] [PDF]
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 F. Nicoletti, R. Di Marco, K. Mangano, F. Patti, E. Reggio, A. Nicoletti, K. Bendtzen, and A. Reggio Increased serum levels of interleukin-18 in patients with multiple sclerosis Neurology, July 24, 2001; 57(2): 342 - 344. [Abstract] [Full Text] [PDF]
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D. Neumann, E. Del Giudice, A. Ciaramella, D. Boraschi, and P. Bossu Lymphocytes from Autoimmune MRL lpr/lpr Mice Are Hyperresponsive to IL-18 and Overexpress the IL-18 Receptor Accessory Chain J. Immunol., March 15, 2001; 166(6): 3757 - 3762. [Abstract] [Full Text] [PDF]
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R. Debets, J. C. Timans, T. Churakowa, S. Zurawski, R. de Waal Malefyt, K. W. Moore, J. S. Abrams, A. O'Garra, J. F. Bazan, and R. A. Kastelein IL-18 Receptors, Their Role in Ligand Binding and Function: Anti-IL-1RAcPL Antibody, a Potent Antagonist of IL-18 J. Immunol., November 1, 2000; 165(9): 4950 - 4956. [Abstract] [Full Text] [PDF]
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F.-D. Shi, K. Takeda, S. Akira, N. Sarvetnick, and H.-G. Ljunggren IL-18 Directs Autoreactive T Cells and Promotes Autodestruction in the Central Nervous System Via Induction of IFN-{gamma} by NK Cells J. Immunol., September 15, 2000; 165(6): 3099 - 3104. [Abstract] [Full Text] [PDF]
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P. I. Karachunski, N. S. Ostlie, C. Monfardini, and B. M. Conti-Fine Absence of IFN-{gamma} or IL-12 Has Different Effects on Experimental Myasthenia Gravis in C57BL/6 Mice J. Immunol., May 15, 2000; 164(10): 5236 - 5244. [Abstract] [Full Text] [PDF]
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S. Nakamura, T. Otani, Y. Ijiri, R. Motoda, M. Kurimoto, and K. Orita IFN-{gamma}-Dependent and -Independent Mechanisms in Adverse Effects Caused by Concomitant Administration of IL-18 and IL-12 J. Immunol., March 15, 2000; 164(6): 3330 - 3336. [Abstract] [Full Text] [PDF]
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 I. A. Schrijver, M.-J. Melief, M. van Meurs, A. R. Companjen, and J. D. Laman Pararosaniline Fixation for Detection of Co-stimulatory Molecules, Cytokines, and Specific Antibody J. Histochem. Cytochem., January 1, 2000; 48(1): 95 - 104. [Abstract] [Full Text]
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B. Siegmund, H.-A. Lehr, G. Fantuzzi, and C. A. Dinarello IL-1beta -converting enzyme (caspase-1) in intestinal inflammation PNAS, November 6, 2001; 98(23): 13249 - 13254. [Abstract] [Full Text] [PDF]
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