| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Third Department of Medicine and
Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan;
Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan; and
§
Departments of Microbiology-Immunology and Pathology, Northwestern University Medical School, Chicago, IL 60611
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
and IFN-
in the spleen cells was decreased, and that of
Th2-derived cytokines such as IL-4 and IL-10 was increased. The delayed
type hypersensitivity and T cell proliferative response specific for
TMEV were decreased by this treatment. These data suggest that IL-12 is
critically involved in the pathogenesis of TMEV-IDD and that Abs to
IL-12 could be a novel therapeutic approach in the clinical treatment
of demyelinating diseases such as human multiple
sclerosis. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Intracerebral infection of susceptible mouse strains with Theiler’s
murine encephalomyelitis virus (TMEV) induces a chronic progressive
demyelinating disease characterized histologically by perivascular
inflammatory cell infiltrates and primary demyelination of the CNS and
clinically by progressive hind limb paralysis (2, 3, 4). TMEV-induced
demyelinating disease is considered an infectious mouse model for MS
because the disease displays similar histopathologic, genetic, and
clinical similarities to human MS (3, 5, 6, 7). Persistent CNS virus
infection in susceptible mouse strains triggers clonal expansion and
differentiation of TMEV-specific, MHC class II-restricted effector DTH
(Th1) cells that are poorly controlled by normal immunoregulatory
mechanisms. Proinflammatory cytokines produced by virus-specific Th1
cells lead to the recruitment, accumulation, and activation of
additional monocytes and macrophages within the CNS that cause
demyelination through a terminal nonspecific bystander response (8, 9).
IFN- plays a particularly key role in the TMEV-IDD inflammatory
response, and a perturbation in the level of this cytokine results in
acceleration of demyelinating disease (10).
IL-12 is a recently discovered cytokine produced predominantly by
macrophages and specific Ag-presenting cells such as dendritic cells
and Langerhans cells (11). This cytokine is required for effective Th1
cell generation (12). One of the central mechanisms by which IL-12
induces differentiation of Th1 cells is its ability to prime T cells
during clonal expansion for high IFN- production (13). IL-12 also
influences the efficacy of Ag presentation by T cells by providing a
costimulatory signal for T cell activation (14, 15). These findings
suggest that IL-12 plays an important role in the induction of clonal
expansion and differentiation of TMEV-specific, MHC Class II-restricted
effector DTH (Th1) cells. In the present study, we examined whether
IL-12 is involved in the pathogenesis of virally induced demyelination
and whether systemic administration of Abs to IL-12 could be an
effective treatment of virus-induced demyelinating disease.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Female pathogen-free SJL/J mice from The Jackson Laboratory (Bar Harbor, Maine) and female BALB/c mice from Japan SLC (Shizuoka, Japan) were housed and cared for in an approved facility, in accordance with the National Institutes of Health guidelines.
Virus
The BeAn 8386 strain of TMEV was propagated in baby hamster kidney-21 cells grown in DMEM supplemented with 7.5% donor calf serum and purified by isopycnic centrifugation on Cs2SO4 gradients as previously described (16).
Monoclonal Abs
The mAbs used in this study were C17.8 and M18/2, rat IgG2a mAbs. C17.8 is a neutralizing rat anti-mouse IL-12 Ab (17, 18). Because M18/2 does not block cell-mediated target cell lysis in vitro (nonneutralizing mAb) (19), we used M18/2 as control mAb. Hybridoma cells that produce these mAbs were cultured in RPMI 1640 supplemented with 10% FBS and 0.1% gentamicin. These cells were injected into nude mice; mAbs harvested as ascites were purified with the use of a protein G affinity column.
Injection of mice with TMEV
For i.c. inoculation of virus, 1.3 x 106 plaque-forming units (PFU) of virus in 30 µl were administered into the right cerebral hemisphere of mice anesthetized with methoxyflurane. This inoculum consistently induces neurologic signs in susceptible mouse strains (20).
Treatment with mAbs
Six- to eight-wk-old female SJL/J mice were separated into
groups (A-G). Group A mice (control) were treated with PBS. In each
experiment, TMEV was injected into SJL/J mice i.c. at day 0. mAbs
(C17.8, M18/2 (non specific rat IgG2a mAb)) were injected i.p. into
mice on days -2, 0, and 4 after i.c. infection with TMEV at induction
phase or 20, 22, and 26 after i.c. infection with TMEV at the effector
phase at a dose of 500 µg at a volume of 100 µl/mouse per
injection. Details of the experimental design are given in Table I
. We performed this mAb treatment
experiment three times. In one experiment, 7 groups of mice were under
investigation (n = 20 for each group). Before
experiments, five mice were blindly selected from each group for
histologic study, and another 5 mice were also blindly selected from
each group for immunologic studies, such as TMEV-specific DTH,
TMEV-specific T cell proliferation assay, and enumeration of
cytokine-producing cells assay. Other mice were clinically observed
until 80 days post i.c. infection (n = 10 for each
group).
|
TMEV-infected mice were examined daily for clinical symptoms of demyelination. Mice were allowed to walk on a polyethylene (Dynalab) walking board and were observed for exhibition of symptoms, including a waddling gait, extensor spasms, paralysis, loss of righting reflex, incontinence, and hunched posture. Neurologic signs were recorded using the following grading system: normal = 0, slight waddling gait = 1, waddling gait = 2, spastic hind limb paralysis = 3, severe hind limb paralysis accompanied by incontinence = 4 (21). These clinical scores have been shown to be indicative of demyelination (2). A clinical score was recorded daily for each mouse in each experiment using this grading system. The mean clinical score for each group of mice on each day was calculated by dividing the sum of all clinical scores of the mice in a given group by the number of mice in that group.
Histology
In each experiment, mice were blindly selected from each group (n = 5 from each group) beforehand for histologic examination and sacrificed on day 40 post i.c. infection. Since we had repeated this experiment three times, 15 mice from each group were histologically examined. Mice were perfused under anesthesia by the intraventricular route with 4% paraformaldehyde in PBS, pH 7.4. Spinal cords were removed, fixed in 4% paraformaldehyde and 2.5% glutaraldehyde in PBS, and embedded in epoxy resin. These epoxy-embedded (1 µm thick) sections were stained with toluidine blue. These sections from 12 segments/mouse were read under light microscopy, and grading was done in a blinded fashion by two independent investigators who were unaware of the treatment each animal had received. The score for inflammation was determined according to the following criteria: 0, none; 1, a few inflammatory cells; 2, numerous scattered cells with an occasional perivascular cuff; 3, many perivascular cuffs; 4 and 5, increasing perivascular infiltration and subarachnoid inflammation. The extent of demyelination was determined according to the following scoring system: 0, no demyelination; 1, a few scattered naked axons; 2, small groups of naked axons; 3, large groups of demyelinated axons with confluent plaques of demyelination (22).
Ag-specific DTH
A 24-h ear swelling assay was used to quantitate delayed type hypersensitivity (DTH) (23). Before experiments, five mice were also blindly selected from each group for immunologic studies. At 36 days post i.c. infection, prechallenge ear thickness of these mice was determined using a Mitutoyo digimatic micrometer (Mitutoyo, Tokyo, Japan). Subsequently, 5 µg of purified TMEV in 10 µl of saline was injected into the dorsal surface of the ear using a Hamilton syringe fitted with a 30-gauge needle. Twenty four hours later, ear thickness was again measured, and the increase in thickness was expressed in units of 10-4 inches. Ear swelling reactions were due to mononuclear cell infiltration and showed typical DTH kinetics (i.e., minimal swelling at 4 h, maximal swelling at 24 to 48 h).
T cell proliferation assay
After DTH measurement, the same mice were sacrificed. Spleen cells were harvested from three animals in each group and pooled. Cells (5 x 105) were cultured in 96-well flat-bottom microculture plates in RPMI 1640 containing 0.5% syngeneic mouse serum, 5 x 10-5 M 2-ME, and antibiotics. Triplicate cultures were stimulated with three different concentrations of UV-inactivated TMEV (0.5, 5, and 10 µg) and were incubated for 72 h in a humidified atmosphere of 5% CO2 and 95% air. Cultures were then pulsed with 1.0 µCi of [3H]dThd and harvested 24 h later. Measurement of [3H] dThd incorporation was determined using a scintillation counter and expressed as cpm. Background proliferation was less than 1/7 of TMEV-specific proliferation.
Anti-TMEV Ab and anti-TMEV subclass titration
TMEV-specific Ab titers were determined using ELISA as described earlier (24), utilizing sera from individual mice. Briefly, 0.3 µg of purified virus was used to coat microtiter plates. A BSA solution (0.3 µg) was also used to coat the plates, to serve as a negative control. Unless otherwise stated, twofold serial diluents of sera starting from a 1:100 (20 x 100) dilution were reacted with the Ags on the microtiter plates and then with goat anti-mouse secondary Ab conjugated with alkaline phosphatase (KPL, Gaithersburg, MD). For anti-TMEV subclass Ab titration, sera were reacted with the Ag on the microplates and then with biotinylated rat monoclonal anti-mouse IgG subclass Ab (Zymed, San Francisco, CA). After the plates were washed, streptavidin-alkaline phosphatase was added to each well, and the plates were incubated. After the plates were again washed, substrate (p-nitrophenyl phosphate) for the enzyme was added, and the enzyme reaction was colorimetrically measured by an ELISA reader at 410 nm. The Ab titers of ELISA represent log2 x 100.
Cytokine assay by ELISA
The concentration of circulating cytokines such as TNF-,
IFN-, IL-4, or IL-10 were measured using commercially available
ELISA kits (Genzyme, Cambridge, MA).
Enumeration of cytokine-producing cells
On day 40 post i.c. infection, spleen cells from the same mice
used for Ag-specific DTH and T cell proliferation assay
(n = 5 from each group) were harvested from animals in
each group. The levels of TNF-, IFN-, IL-4, or IL-10-producing
spleen cells were examined using enzyme-linked immunospot assay
(ELISPOT). The original reverse ELISPOT assay (25) was
modified by using nitrocellulose membrane (Bio-Rad, Hercules, CA).
Wells were filled (50 µl/well) with monoclonal hamster
anti-murine TNF-, IFN-, IL-4, or IL-10 mAbs (Genzyme) at a
concentration of 10 µg/ml in 0.5% BSA in PBS overnight at 4°C.
Unabsorbed Abs were removed, and wells were washed with PBS. The plates
were then blocked with 1% BLOTTO (nonfat dry milk) for 2 h at
37°C. The outer surface of the nitrocellulose membrane was carefully
dried. Spleen cells (1 x 105/well) in the culture
medium (RPMI 1640 supplemented with 10% FBS and 0.1% gentamicin) were
dispensed among individual wells (100 µl/well). Plates were then
incubated for 48 h at 37°C in a humidified, 5% CO2
atmosphere and were washed three times with Tris-buffered saline with
Tween 20 (TBST). Fifty microliters of a 1:250 dilution of polyclonal
rabbit anti-murine TNF-, IFN-, IL-4, or IL-10 Abs (Genzyme)
was added to each well followed by incubation for 2 h at 37°C.
Plates were washed with TBST again and were treated with 50 µl
of 500 µg/ml alkaline phosphatase-conjugated goat
anti-rabbit IgG (KPL) for 2 h at 37°C. After another washing
with TBST, cytokines secreted by single cells were visualized by adding
a mixture of nitro-blue tetrazolium and 5-bromo-4-chloro-3-indole
phosphate (Life Technologies, Inc., Grand Island, NY). The color
reaction of the enzyme was halted after 30 min by washing with water,
and spots were enumerated under x40 magnification.
Statistical analysis
Clinical scores were analyzed using the Mann-Whitney U test; other results were statistically evaluated using the Student t test (StatView program, Abacus Concepts, Berkeley CA). A p value of <0.05 was considered statistically significant.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The results of experiments are summarized in Fig. 1 and Table II. Control animals (group A, nontreated;
Groups C, E, and G, treated with control Ab M18/2) showed the typical
disease course of TMEV-IDD. On day 40, about 70% of mice showed
clinical signs such as waddling gait, extensor spasm, and hind leg
paralysis; mean clinical scores were 2.7 in group A, 2.4 in group C,
2.5 in group E, and 2.8 in group G, respectively. On day 80, all mice
from groups A, C, E, and G developed TMEV-IDD; the mean clinical scores
were 3.5 in group A, 3.6 in group C, 3.7 in group E, and 3.8 in group
G, respectively. In contrast, on day 40, no mice treated with
anti-IL-12 mAb in the effector phase (group D) had clinical signs
(the mean clinical score was 0), while 33% of mice treated with
anti-IL-12 mAb in the induction phase (group B) had clinical signs,
with a mean clinical score of 0.9. Fifteen percent of mice treated with
anti-IL-12 mAb in both induction and effector phases (group F) had
clinical signs; the mean clinical score was 0.6. These results
demonstrate that clinical symptoms of demyelinating disease are
significantly suppressed (footnotes a, d, and e,
p < 0.01; footnote b, p < 0.005) in
animals treated with anti-IL-12 mAb (groups B, D, and F), as
compared with those in control groups (groups A, C, E, and G). On day
80 post i.c. infection, as shown by the mean clinical score, severities
of neurologic signs were significantly lower (footnote c,
p < 0.01) in mice treated with neutralizing anti IL-12
mAb in the effector phase (group D) than in control groups (groups A,
C, E, and G). However, we could not detect any significant differences
in other treatment groups (groups B and F).
|
|
A). The inflammation scores
and demyelination scores were significantly lower in mice from groups
B, D, F (footnotes d and e, p < 0.01) than those from
control groups (Fig. 2B; Table II
).
|
Virus-specific DTH, as measured by the ear-swelling assay, is
known to correlate strongly with susceptibility to TMEV (23); DTH has
been shown to be mediated by the Th1 lymphocyte subset (26). To compare
the clinical signs and the level of TMEV-specific DTH, we assessed the
level of DTH 36 days after viral infection in mice selected for
immunologic examination (n = 5 from each group). The
levels of DTH at 36 days postinfection in mice of control groups
(groups A, C, E, and G) were increased at almost the same levels
previously reported (27). Conversely, they were significantly lower
(p < 0.01) in mice treated with neutralizing
anti IL-12 Abs (groups B, D, and F). These results show that
administration of these mAbs inhibits the level of TMEV-specific DTH
(Fig. 3A). We also examined
the level of TMEV-specific DTH at 78 days post i.c. infection. Mice
were chosen blindly from each group (n = 5 from each
group). In this stage, the DTH level was decreased only in mice treated
with anti-IL-12 during effector phase (group D).
|
T cell proliferative responses have been used frequently to assess
the level of response of CD4+ T helper cells to the virus
(28). After DTH measurement, the same mice were sacrificed on day 40
post i.c. infection. Spleen cells were taken out and used for
examination for TMEV-specific T cell proliferation and ELISPOT assay.
When the TMEV-specific T cell proliferative responses of mice treated
with neutralizing anti-IL-12 Abs were compared with those of
control groups, we observed a distinct difference (Fig. 3B).
These results may indicate that administration of anti-IL-12 mAb
inhibits the ability of TMEV-specific T cell proliferation of
TMEV-specific mouse spleen cells or inhibits the generation of
TMEV-specific T cells at the precursor level.
We also examined the level of TMEV-specific T cell proliferation at 80 days post i.c. infection. The same mice that were used for TMEV-specific DTH at 78 days post i.c. infection were examined (n = 5 for each group). In this stage, TMEV-specific T cell proliferative response was decreased only in mice treated with anti-IL-12 during effector phase (group D).
TMEV-specific Ab responses
We examined the Ab responses in the experimental groups of mice to
determine a possible effect of anti-IL-12 Abs on the production of
TMEV-specific Abs. Sera were taken from all mice of each group
(n = 10 from each group) on day 56 and day 80 post i.c.
infection. On day 56, there was no significant difference in the
TMEV-specific Ab levels among the six groups. We also explored whether
Ab isotypes were affected. We detected IgG1 and IgG2b Abs in all
groups, with no significant difference among the 7 groups. We also
detected IgG2a Ab in control groups (groups A, C, E, and G), but not in
the anti-IL-12 mAb-neutralizing Ab treated groups (groups B, D, and
F) (Fig. 4). These results indicate that,
despite an unaltered level of total Abs, there is a lack of IgG2a
component in animals treated with anti-IL-12-neutralizing Abs.
|
Assessments of cytokine production
After DTH measurement, the same mice (n = 5 from
each group) were sacrificed on day 40 post i.c. infection. Spleen cells
were taken out and used for ELISPOT assay. We could not detect any
TNF-, IFN-, IL-4, or IL-10 in the sera from animals of any group.
The levels of cytokine-producing cells in the spleens of animals were
also assessed using the ELISPOT method. TNF- production by spleen
cells from mice treated with neutralizing Abs to IL-12 was
significantly suppressed (p < 0.01) compared
with control groups (Fig. 5A).
IFN- production by spleen cells from mice treated with
anti-IL-12-neutralizing Abs was also significantly suppressed
(p < 0.01) compared with control groups (Fig. 5B). Production of IL-4 and IL-10 by spleen cells from mice
treated with anti-IL-12-neutralizing Abs was significantly increased
(p < 0.01) compared with control groups (Fig. 5, C and D). Since we could not detect any
cytokine spots in uninfected mice and uninfected mice treated with
anti-IL-12 mAb, these results suggest that production of Th1
cell-derived, inflammatory cytokines was down-regulated and Th2
cell-derived cytokine production was somewhat up-regulated in mice
treated with anti-IL-12-neutralizing Abs.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
production (12, 36). Th1 cells
produce IFN- and TNF-ß, which activate macrophages and are
involved in DTH reactions. IL-12 is critically involved in the
development of naive Th cells into Th1 cells (37). As a result of this
ability to drive T cell responses to the Th1 phenotype, IL-12 has been
shown to have exacerbating effects on several autoimmune disease
models. Administration of IL-12 induced a) earlier onset of
insulin-dependent diabetes mellitus in female NOD mice (38); b) more
severe and prolonged disease in adoptively transferred experimental
autoimmune encephalomyelitis (EAE), another animal model of human MS
(39); c) destructive collagen-induced arthritis (40); and d)
accelerated glomerulonephritis in young MRL/lpr mice (41).
The suppressive effects of this mAb on several immune mediated diseases
have also been reported. Monoclonal Abs against IL-12 have been shown
to abrogate experimental chronic inflammatory colitis in mice, which is
an animal model of Crohn’s disease, mediated by Th1 T cells (42).
Furthermore, Abs to IL-12 have been shown to prevent EAE. (39). EAE is
an autoimmune disease of the CNS that can be induced in susceptible
strains of mice by immunization with CNS myelin Ags, or, alternatively,
the disease can be passively transferred to naive susceptible
mice via Ag-specific CD4+Th1 cells (43). In EAE, treatment
of mice with Ab to murine IL-12 after cell transfer completely prevents
paralysis (39). In our present study, treatment with
anti-IL-12-neutralizing Abs significantly suppressed the occurrence
and severity of demyelinating disease, but not completely.
Virus-specific DTH and T cell proliferative responses were also
decreased by this treatment. Our ELISPOT study additionally showed that
in anti-IL-12 mAb-treated groups, spleen cell production of TNF-
and IFN- was decreased and production of IL-4 and IL-10 was
increased, compared with nontreated groups and nonspecific IgG-treated
groups. Our data suggest that treatment with anti-IL-12 mAbs
shifted Th1-Th2 balance to Th2 dominance. Decreased production of
Th1-derived cytokines such as TNF- and IFN- and increased
production of Th2-derived cytokines such as IL-4 and IL-10 might
suppress CD4+ virus-specific DTH cells, which correlated
well with the disease severity of TMEV-IDD, and suppressed
demyelinating disease. In Ab study, though titers of virus-specific Abs
were increased in mice from all groups, IgG2a isotypes were decreased
in anti-IL-12 mAb-treated groups. Since IgG2a Abs are produced by
the stimulation of Th1 cells, decreased levels of virus-specific IgG2a
Abs suggest the suppression of Th1 cells. Taken together, induction of
encephalitogenic CD4+ Th1 cells might be suppressed by the
treatment of anti-IL-12 mAbs. The suppressive effect on
demyelinating disease was prominent in groups of mice treated with anti
IL-12 mAbs during the effector phase. During this period, induction of
encephalitogenic CD4+ Th1 cells is prominent; inhibition of
Th1 cells by anti IL-12 treatment may be effective for the suppression
of demyelinating disease. In contrast, in anti-IL-12 mAb treatment
during the induction or induction and effector phases, though on day 40
post i.c. infection, the suppressive effect was prominent; on day 80,
there was no significant difference compared with control treated
groups. IL-12 plays an important role in immune response to viral
infections, as seen in lymphocytic choriomeningitis virus infection of
C57BL/6 mice, an acute viral infection model of the CNS, in which
administration of low dose IL-12 is protective and high dose IL-12 is
detrimental (44). IL-12 injection at the time of vaccination stimulates
an antiviral type 1 cytokine response and increases immunity against a
neurotrophic herpes virus infection of mice (44). IL-12 treatment
decreases viral titer of vesicular stomatitis virus (VSV)-infected mice
and inhibits productive vesicular stomatitis virus infection of the CNS
(46). These findings suggest that, in the induction phase,
anti-IL-12 treatment may reduce the antiviral activity of Th1
cells. Whether anti-IL-12 mAb treatment had any effect on virus
replication or persistence in the CNS would be a primary factor in
assessing the usefulness of this anti-IL-12 therapy. Possibly, the
anti-IL-12 therapy in the induction phase may cause dramatic
elevations in CNS virus that would augment a direct mechanism of
infectious pathogenesis. Then, increased levels of virus in CNS may
lead the progression of demyelinating disease. Change of immunologic
parameters such as TMEV-specific DTH, T cell proliferation, and Ab
subtype from day 40 to day 80 may support this hypothesis.
Alternatively, the anti-IL-12 therapy only in the effector phase
may have little effect on CNS virus levels/persistence, and the
inhibition of encephalitogenic Th1 cells may lead the suppression of
TMEV-IDD. Finally, since methods of clinical therapy of MS are still
incomplete, it is important to examine the possibility of
anti-cytokine therapy, including mAb to IL-12.
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Chang-Sung Koh, Third Department of Medicine (Neurology), Shinshu University School of Medicine, 3–1-1 Asahi, Matsumoto 390-8621, Japan. E-mail address:
3 Abbreviations used in this paper: MS, multiple sclerosis; TMEV-IDD, Theiler’s murine encephalomyelitis virus-induced demyelinating disease; EAE, experimental autoimmune encephalomyelitis; DTH, delayed-type hypersensitivity; CNS, central nervous system; i.c., intracerebral; ELISPOT, enzyme-linked immunospot.
Received for publication April 8, 1998. Accepted for publication July 14, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
results in acceleration of Theiler’s virus-induced demyelinating disease. Neuroimmunology 55:143.[Medline]
(IFN-) production during differentiation of human T helper (Th) cells and transient IFN-g production in established Th2 cell clones. J. Exp. Med. 179:1273. production by mouse T helper clones that are unresponsive to B7 costimulation. J. Exp. Med. 180:223. production and lethality in LPS-induced shock in mice. Eur. J. Immunol. 25:672.[Medline]
- and ß-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and MAC-1. J. Exp. Med. 158:586.
gt11 system: different patterns of antibody reactivity among different mouse strains. Neuroimmunology 27:173.[Medline]
-receptor-deficient mice. J. Immunol. 155:2525.[Abstract]
This article has been cited by other articles:
|
|
 |
|
  P. Kapil, R. Atkinson, C. Ramakrishna, D. J. Cua, C. C. Bergmann, and S. A. Stohlman Interleukin-12 (IL-12), but Not IL-23, Deficiency Ameliorates Viral Encephalitis without Affecting Viral Control J. Virol., June 15, 2009; 83(12): 5978 - 5986. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Li, L. Fu, D. M. Gonzales, and E. Lavi Coronavirus Neurovirulence Correlates with the Ability of the Virus To Induce Proinflammatory Cytokine Signals from Astrocytes and Microglia J. Virol., April 1, 2004; 78(7): 3398 - 3406. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. L. Oleszak, J. R. Chang, H. Friedman, C. D. Katsetos, and C. D. Platsoucas Theiler's Virus Infection: a Model for Multiple Sclerosis Clin. Microbiol. Rev., January 1, 2004; 17(1): 174 - 207. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Weeber, R. Vos, H. Klein, L. T W de Jong-van den Berg, A. R Aronson, and G. Molema Generating Hypotheses by Discovering Implicit Associations in the Literature: A Case Report of a Search for New Potential Therapeutic Uses for Thalidomide JAMIA, May 1, 2003; 10(3): 252 - 259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Weeber, R. Vos, H. Klein, L. T.W. de Jong-van den Berg, A. R. Aronson, and G. Molema Generating Hypotheses by Discovering Implicit Associations in the Literature: A Case Report of a Search for New Potential Therapeutic Uses for Thalidomide J. Am. Med. Inform. Assoc., May 1, 2003; 10(3): 252 - 259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Palma, R. L. Yauch, H.-K. Kang, H.-G. Lee, and B. S. Kim Preferential Induction of IL-10 in APC Correlates with a Switch from Th1 to Th2 Response Following Infection with a Low Pathogenic Variant of Theiler's Virus J. Immunol., April 15, 2002; 168(8): 4221 - 4230. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
ear swelling in units of 10-4
inches ± SEM. Average background ear-swelling responses (from
mock-infected mice) were subtracted from the individual specific
ear-swelling responses. The average background ear-swelling was
3.2 ± 1.2. The responses of groups B, D, and F were significantly
lower (*, p < 0.01) than those of other groups.
B, The level of TMEV-specific T cell proliferative
response was significantly lower in mice treated with mAbs to IL-12
than control groups treated with either PBS (none) or a nonspecific IgG
mAb (*, p < 0.01). On day 40 post viral
infection, the same mice from DTH study were sacrificed and examined
for TMEV-specific T cell proliferative response. Spleen cells (5
x 105) were cultured with 10 µg of intact UV-inactivated
TMEV for 72 h and then pulsed with 1 µCi of
[3H]TdR for the final 24 h. Data represent specific
proliferation minus nonspecific proliferation of spleen cells. Results
are expressed as mean cpm ± SEM from triplicate cultures.
Treatments were as follows: group A, none (PBS) treated; group B,
anti-IL-12 mAb (induction); group C, nonspecific IgG mAb
(induction); group D, anti-IL-12 mAb (effector); group E,
nonspecific IgG mAb (effector); group F, anti-IL-12 mAb (induction
+ effector); and group G, nonspecific IgG mAb (induction +
effector).
