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Departments of
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Neurology and Molecular Microbiology,
Immunology, and
Pathology, University of Southern California, Los Angeles, CA 90033
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In this report we examined the requirement for CD4+ T cells in virus clearance from the parenchyma of the CNS following infection by the neurotropic JHM strain of mouse hepatitis virus (JHMV). Analysis of immune infiltrates within the CNS of rodents infected with JHMV support the importance of CTL in the antiviral response by demonstrating that CD8+ T cell accumulation coincides with declining virus titers (25, 26). The protective role of CTL during JHMV infection of the CNS was recently confirmed by the effects of adoptively transferred CTL specific for the JHMV nucleocapsid (N) protein into lethally infected recipients (27). N-specific CTL mediated protection via partial elimination of CNS virus replication. Whereas CTL suppressed virus replication in astrocytes and microglia (27), they appeared to have had little or no direct effect on virus replication in oligodendroglia.
Induction of CTL and subsequent clearance of many viruses are independent of CD4+ T cells (1, 2, 18, 28, 29). For example, vaccinia virus, influenza virus, and LCMV are cleared via a vigorous CTL response in CD4-depleted mice or in mice in which the CD4+ T cell response has been eliminated by gene disruption (29). By contrast, Rauscher leukemia virus infection and Japanese encephalitis virus infection of the CNS require both CTL and CD4+ T cells for protection (30, 31). CD8+ CTL-mediated clearance of JHMV from the CNS also appears to be dependent on CD4+ T cell-mediated help. This idea is based on the demonstration that adoptive transfer of a virus-specific CD4+ T cell population mediated protection and viral clearance from the CNS in an MHC class I-dependent manner (32). It has been further supported by the absence of JHMV clearance from mice depleted of CD4+ T cells (33) or mice in which class II has been eliminated by gene deletion (34) and the demonstrations that both virus-specific CD4+ and CD8+ T cell clones protect from lethal JHMV infections (35, 36, 37). These data suggest that the host’s immune response to CNS infection by JHMV includes a CD4+ T cell component necessary for CTL function. Whether this is an intrinsic property of a virus that exhibits cytotoxicity in vitro, but little or no direct cytopathology in vivo, or whether this is due to the relatively immune-privileged target organ of infection is not clear. In support of the idea that the site may play a role in the regulation of CTL function, excellent CTL activity can be detected in mononuclear cells isolated directly from the CNS during acute JHMV infection (38, 39, 40, 41). However, there appears to be little or no JHMV-specific CTL activity in peripheral organs during JHMV infection (38, 40, 41).
This report confirms that the ability of CTL to reduce JHMV replication in the CNS is dependent on CD4+ T cells. Mice depleted of CD4+ T cells show decreased CD8+ T cell cytolytic activity following infection, suggesting that CD4+ T cells contribute to but are not required for the development of CTL effector function. Adoptive transfers of BrdU-labeled CTL activated in vitro into infected CD4+ T cell-deficient recipients showed that entry into the CNS parenchyma was not inhibited, although the number of cells within the CNS that expressed CD8 was reduced. These data were reconciled by demonstrating the presence of increased numbers of apoptotic lymphocytes within the CNS of CD4-depleted mice. These findings indicate that CD4+ T cells are required for the maintenance of CD8+ T cell effector function within the CNS, but are not an absolute requirement for either CTL induction or trafficking of CD8+ T cells into the parenchyma during acute CNS viral infection.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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BALB/c By (H-2d), mice were obtained from The
Jackson Laboratory (Bar Harbor, ME) and housed in isolator cages.
Donors were immunized as previously described (27) using 1 x
106 plaque-forming units JHMV injected i.p. Recipient and
control mice were infected intracranially with 100 plaque-forming units
of JHMV in 30 µl of PBS. This inoculum is uniformly fatal within 7 to
9 days postinfection. Recipients were depleted of CD4+ T
cells by i.p. injection of 200 µg of anti-CD4 mAb GK1.5 at -2,
0, and +2 days relative to infection as previously described (33). Mice
depleted of CD8+ T cells were injected on the same schedule
with 200 µg of anti-CD8 mAb 2.43. Both regimens resulted in
98% depletion of their respective phenotypes, as analyzed by flow
cytometry (see below), 4 days after the final Ab injection.
Viruses and cell lines
The DM strain, a plaque-purified isolate derived from a suckling mouse brain pool that has plaque morphology and pathogenesis consistent with those of parental JHMV (42) was propagated and plaque assayed using the murine DBT astrocytoma cell line as previously described (27, 39). Virus titers were determined by homogenization of one-half the brain in 2.0 ml of Dulbecco’s PBS, pH 7.4, using Tenbroeck tissue homogenizers. The remainder was used for histopathologic analysis (see below). Following centrifugation at 1500 x g for 10 min at 4°C, supernatants were assayed immediately or were frozen at -70°C. Virus titers were determined by plaque assay using monolayers of DBT cells as previously described (27). The data presented are the average titers of groups of at least three mice per point.
Induction and transfer of CTL effectors
Spleen cell suspensions were prepared from mice immunized 3 to 8
wk earlier. Spleen cells (1 x 108) were cultured
for 6 days at 37°C in 40 ml of RPMI 1640 medium supplemented with
10% FBS (Gemini Bioproducts, Calabasas, CA), 2 mM glutamine, 25
µg/ml gentamicin, 1 mM sodium pyruvate, 5 x 10-5 M
ß2-ME, and nonessential amino acids (RPMI complete) plus 10% rat Con
A supernatant (RCS) containing 25 mM 
-methylmannopyranoside and 1
µM pN318–335 peptide as previously described (27, 43).
The 18-mer peptide containing the CTL epitope was used in most
experiments due to enhanced solubility compared with that of the
optimal 9-mer peptide. Viable cells were purified by centrifugation
onto Lympholyte M (Accurate Chemicals, Westbury, NY) cushions and
washed twice before transfer into naive recipients. Total cells (2
x 107) or CD8+ T cells (1 x
107) purified by positive selection using magnetic beads
(see below) were transferred i.v. either 4 to 6 h before infection
or at 2 days postinfection for BrdU-labeled cells (day 3 of infection).
Recipients were challenged with JHMV within 6 h of adoptive
transfer. To label CD8+ T cells for analysis of trafficking
into the CNS, 50 nM BrdU (Sigma Chemical Co., St. Louis, MO) was added
during in vitro expansion of memory cells for the final 48-h incubation
(15). BrdU-labeled CD8+ T cells were purified by positive
selection, as described below, before i.v. transfer. Inclusion of 50 nM
BrdU in the culture medium for 48 h did not alter cytolytic
activity compared with that in untreated cultures.
Flow cytometry and T cell purification
Cell surface expression following in vitro activation was examined using rat anti-CD4 (L3T4), rat anti-CD8 (53-5.8), rat anti-CD11a (M17/4), and rat anti-CD62L (MEL-14; PharMingen, San Diego, CA) followed by goat anti-rat FITC-F(ab')2 Ab (Caltag, South San Francisco, CA). Cells were labeled, washed, resuspended in PBS containing 0.1% formaldehyde, and analyzed on a FACStar (Becton Dickinson, Mountain View, CA). Ab plus C-mediated depletions were conducted as previously described (27), using rat anti-CD4 (mAb RL172.4) and anti-CD8 (mAb 31M).
CD8+ T cells were purified using MACS Microbeads conjugated
to anti-CD8 mAb (clone 53-6.7; Miltenyi Biotec, Auburn, CA).
Peptide-stimulated cells (1.5 x 108) were incubated
for 40 min at 4°C with 100 µl of a colloidal microbead solution.
The cell suspension was passed over a saturated steel wool column
equilibrated with ice cold-PBS containing 0.5% BSA (PBS/BSA) attached
to a MACS magnet (Miltenyi Biotec). The unabsorbed CD8-
fraction was collected by eluting the column with 5 vol of chilled
PBS/BSA at a slow flow rate regulated by a 23-gauge needle. The
absorbed CD8+ fraction was collected by removing the magnet
and eluting the cells with PBS/BSA. Typically, >20% of the cells
applied to the column were recovered in the CD8+ fraction.
Purity was 98% as determined by flow cytometry.
CD4+ T cells from cultures stimulated in vitro with peptide pN318–335 were purified using MACS microbeads conjugated with anti-CD4 mAb (GK1.5; Miltenyi Biotec) as described above. For proliferation, CD4+ cells (1 x 105) were cultured in complete RPMI 1640 medium supplemented with 7 x 105 irradiated syngeneic spleen cells and 1% syngeneic mouse serum in 96-well plates. Cells were stimulated with lysates of UV-inactivated, JHMV-infected, DBT cells (36) or with 0.2 to 20 nM peptides (pN318–335 or pN318–326) for 72 h at 37°C and pulsed with 2 µCi of [3H]dThd (ICN Radiochemicals, Irvine, CA)/well for the last 16 h of culture. Incorporation was measured by liquid scintillation spectroscopy.
Cytotoxicity assay
Spleen cell suspensions (1 x 108)
prepared from infected normal and CD4-depleted mice were cultured for 6
days at 37°C with 0.1 µM pN318–335 in 40 ml of
complete RPMI 1640 medium supplemented with 10% FBS (Gemini
Bioproducts). Cytolytic activity was measured in a 4-h 51Cr
release assay as previously described (27, 43). J774.1
(H-2d) target cells were washed and labeled with 100 µCi
of Na51CrO4 (New England Nuclear, Boston, MA).
Washed target cells were preincubated for 15 min at 37°C with 0.5 to
1.0 µM peptide and added at a concentration of 1 x
104 in a 100-µl volume. Data are expressed as the percent
specific release, defined as (experimental release - spontaneous
release)/total (detergent release - spontaneous release). Maximum
spontaneous release values were 
20% of the total release values.
Histology and cell trafficking
For histopathologic analysis mice were killed by CO2 asphyxiation. Brains were removed and bisected in the midcoronal plane. Brains and spinal cords were fixed for 3 h in Clark’s solution (75% ethanol and 25% glacial acetic acid) and embedded in paraffin. Sections were stained with either hematoxylin and eosin or luxol fast blue for routine examination. The distribution of JHMV Ag was examined using immunoperoxidase staining (Vectastain-ABC kit, Vector Laboratories, Burlingame, CA) and anti-JHMV mAb J.3.3 specific for the carboxyl terminus of the N protein (44). The distribution of BrdU+ cells was determined on paraffin-embedded sections by the immunoperoxidase method (Vectastains ABC Kit) using mouse mAb BU-33 (Sigma). To identify CD4+ and CD8+ T cells, immunoperoxidase staining was performed on acetone-fixed frozen sections using rat anti-CD4+ (L3T4, PharMingen) and rat anti-CD8+ (Ly-2, PharMingen). Anti-CD4+ and CD8+ Ab were detected with biotinylated rabbit anti-rat Ab preabsorbed with mouse serum (Vector Laboratories). Visualization was achieved using the ABC kit with 3-amino-9-ethylcarbazole as chromogen. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling was used to examine the distribution of apoptotic cells according to the supplier’s instructions (Oncor, Gaithersburg, MD).
Quantitation of lymphoid cells identified as being CD4+, CD8+, or BrdU+ was performed on immunoperoxidase-stained tissue sections counterstained with hematoxylin. Brain sections were cut in the midsagittal plane of one hemisphere and included olfactory cortex, hippocampus, and cerebellum. All positively labeled cells in the section were counted using a x40 objective, with separate computations for subarachnoid, perivascular, and parenchymal regions. Apoptotic cells were counted in a similar manner, except that the location of the cells was not differentiated since the vast majority of apoptotic cells were intraparenchymal in location.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Adoptive transfer of virus-specific CTL derived from
JHMV-immunized donors expanded in vitro with either N-derived peptide
pN318–335 or pN318–326 mediates MHC
class I-dependent protection from an otherwise lethal infection via the
reduction of JHMV replication within the CNS (27). Analysis of the
transferred population by flow cytometry showed the presence of
approximately equal numbers of activated (MEL-14low)
CD4+ and CD11ahigh CD8+ T cells. To
determine whether activated CD4+ T cells were specific for
the pN318–335 peptide, CD4+ cells were
purified after 6 days in vitro stimulation with peptide and tested for
proliferation to the peptides containing the JHMV N-protein CTL epitope
(43). No stimulation was detected using either the 18-mer
pN318–335 peptide or the 9-mer pN318–326
peptide (data not shown). Minimal JHMV-specific proliferation
(stimulation index = 
3) was detected, suggesting
non-Ag-specific IL-2-driven in vitro expansion of the CD4+
T cells. Although an N-specific proliferative response has previously
been demonstrated in BALB/c mice (45), these data indicate that neither
the optimal 9-mer nor the larger 18-mer peptide contains a
JHMV-specific, H-2d-restricted, CD4+ T cell
epitope. However, the presence of activated CD4+ T cells
suggested the possibility that this population might participate in
clearance of JHMV from the CNS (35, 36, 37). To test whether the
CD4+ T cells derived following in vitro activation played a
role in reduction, CD8+ T cells were either depleted by mAb
plus C or positively selected by magnetic bead purification and
transferred to JHMV-infected recipients. Depletion of CD8+
T cells eliminated the majority, but not all, of the antiviral activity
present in the CNS (Table I
). Cells
treated with C only retained the ability to reduce CNS virus
replication. These data are consistent with previous reports suggesting
that CD4+ T cells participate in the clearance of JHMV
(35, 36, 37). By contrast, the transfer of 1 x 107
affinity-purified CD8+ T cells was as effective as transfer
of 2 x 107 unfractionated cells containing
approximately 50% activated CD8+ T cells (Table I). These
data confirm that CD8+ T cells are a major effector
population mediating the partial reduction of virus replication within
MHC class I-positive CNS cells of JHMV-infected mice (27). Furthermore,
these data indicate that the CD4+ T cells activated in
vitro are not as effective as CTL at reducing virus replication within
the CNS; however, CD4+ T cells activated in situ may
provide protection (35, 36, 37) by an as yet unknown mechanism(s), possibly
via secretion of IFN-
or Fas/Fas ligand-mediated cytolysis.
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The protection afforded by JHMV-specific CD4+ T
cell clones requires recipients with intact immune responses (35). In
vitro activated JHMV-specific CTL were therefore transferred into
untreated and immunosuppressed recipients to determine whether
CTL-mediated virus clearance requires participation of the host’s
immune response. A single dose of irradiation was administered to
recipients on the same day as the i.v. transfer of 2.5 x
107 splenocytes derived by in vitro stimulation of memory
cells with pN peptide. In contrast to untreated recipients,
immunosuppression prevented CTL-mediated reduction of JHMV within the
CNS regardless of whether the CTL were transferred i.v. or
intracranially (Table II). In addition,
immunosuppressed recipients were not protected from fatal disease (data
not shown), similar to the prevention of CD4+ T
cell-mediated protection by immunosuppression (36). The inability of
CD8+ T cells to mediate complete viral clearance from the
CNS (27) or protect immunosuppressed recipients (Table II) suggested
the participation of additional components of the host’s immune
response that were activated during infection.
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). By
contrast, no viral clearance from the CNS was detected in CTL
recipients depleted of CD4+ T cells, nor were they
protected from lethal disease (Table III). These data suggest that
CD4+ T cells provide a critical function during JHMV
infection other than providing help during CTL induction. Depletion of
either CD4+ or CD8+ T cells inhibits JHMV
clearance from the CNS (33), which suggested that CD4+ T
cells are required during induction of JHMV-specific CTL (32, 33). To
determine whether the induction of JHMV-specific CD8+ T
cells is dependent on CD4+ T cells, untreated and
CD4-depleted mice were infected with JHMV, and the cytolytic activity
of splenocytes was determined 6 days postinfection following
pN318–335 peptide restimulation in vitro. Restimulation is
required as no cytolytic activity ex vivo is detectable in the spleen
or cervical lymph nodes during acute JHMV infection (38, 41). CTL
specific for the pN epitope were primed in the absence of
CD4+ T cells (Fig. 1).
However, 16-fold more cells were required to achieve half-maximal
killing (E:T cell ratio = 100:1) compared with cells from
untreated mice (E:T cell ratio = 6:1). The poor cytolytic
activity of CTL derived from infected CD4-depleted donors was overcome
by the addition of exogenous IL-2 in the form of RCS during in vitro
incubation (Fig. 1). These data suggest that CD4+ T cells,
although not required for CTL induction following JHMV infection, play
a role in the development of CD8+ T cell effector function,
presumably by providing help in the form of IL-2 secretion.
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To examine the effects of CD4 and CD8 T cell depletion on the
immunopathology of JHMV infection, mice were depleted of either
CD4+ or CD8+ T cells as previously
described (33). Both treatments were >98% effective as determined by
flow cytometric analysis of splenic populations and inhibited the
clearance of JHMV from the CNS (data not shown). Mice depleted of
either CD4+ or CD8+ T cells showed reduced, but
not absent, mononuclear cell infiltrations and increased numbers of
viral Ag-positive cells compared with untreated controls (Fig. 2), consistent with analysis of mice
rendered CD4 and CD8 deficient by gene ablation (34). These data
suggested the possibility that inhibition of CTL-mediated virus
clearance from the CNS in CD4-depleted recipients was not due to
inhibition of CTL trafficking into the CNS. To examine this
possibility, BrdU was incorporated into pN-specific CTL by incubation
with 50 nM BrdU for the final 48 h of a 6-day in vitro culture
(15). Positively selected CD8+ T cells were transferred to
both untreated and CD4-depleted recipients at 3 days postinfection
(15), and the recipients were killed 48 h later. This time point
was chosen because preliminary experiments showed a significant
reduction of labeled cells within the CNS >2 days after the transfer,
consistent with previous results examining the trafficking of
BrdU-labeled CTL into the livers of transgenic recipients expressing Ag
(15). No differences in BrdU+ cells were detected in the
lungs of recipients, suggesting that the efficacy of the i.v. transfers
was similar in all recipients (data not shown). No difference in either
the number or the distribution of BrdU+ in vitro activated
CTL was detected in the CNS of JHMV-infected recipients when comparing
the untreated and CD4-depleted groups (Fig. 3).
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and 5a). By contrast,
approximately 50% of CD8+ T cells are found within the
brain parenchyma (Figs. 4 and 5b). Consistent with
previous results (13), few T cells are undergoing apoptosis within the
CNS of JHMV-infected mice, and the majority of these are localized
within the parenchyma (Fig. 5c). In CD4-depleted
mice, no CD4+ T cells could be detected in the CNS by
immunohistochemistry, consistent with the 98% depletion of splenic
CD4+ T cells following GK1.5 mAb treatment. The number of
CD8+ T cells infiltrating the CNS was reduced to
approximately 50% the number recruited into the CNS of mice with a
normal complement of CD4+ T cells. Although the overall
number of cells that stain for CD8 was reduced, the percentage of
CD8+ cells within the parenchyma was approximately the same
in the CD4-depleted and untreated groups (Fig. 4). However, the number
of apoptotic cells in CD4-depleted mice increased approximately
threefold (Fig. 5). Although a few apoptotic neurons were detected in
the CD4-depleted mice, the vast majority of the cells undergoing
apoptosis had the morphologic appearance of lymphocytes, consistent
with the near absence of apoptotic cells in the CNS of infected,
immunosuppressed mice (13). Although the identity of the apoptotic
cells could not be established with certainty, morphologic
characteristics, the absence of CD4+ T cells in these mice,
and the predominant perivascular distribution suggest that these cells
largely represent the CD8+ population. These results
suggest that activated CTL traffic normally into the CNS of
CD4-depleted recipients. By contrast, during infection of CD4-deficient
mice, fewer CD8+ T cells were identified in the CNS. The
reduction in CD8+ T cells appears to be due to increased
apoptosis of this population in the absence of CD4+ T
cells.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Whether the increased apoptosis is due to the absence of a tropic factor(s) required for CTL survival or increased Ag load within the CNS of CD4-depleted mice is currently under investigation. However, reduced CTL activity in splenocytes from CD4-depleted infected mice and the recovery of cytolytic activity by IL-2 support its role in enhancing CTL survival and optimal induction of effector function (4, 23, 24). These data suggest that increased apoptotic lymphocytes within the CNS of CD4-depleted recipients may reflect depletion of cytokines or growth factors provided by CD4+ T cells. Indeed, CTL induction is independent of IL-2 (23); however, expression of cytolytic activity is supported by an autocrine loop of IL-2 secreted by activated CD8+ T cells (18, 19, 23). Our data support the idea that CD8+ T cells within the CNS parenchyma may be unable to induce or maintain lytic function due to a defect in the autocrine IL-2 secretion supplemented by the CD4+ T cell population. However, the present data cannot exclude the possibility that the effect on CTL activity is indirect and derived from other cytokines or is a factor(s) derived from endogenous CNS cells (46, 51, 55). For example, CD4+ T cells may secrete or induce the secretion of anti-inflammatory cytokines, such as IL-4, IL-7, IL-10, and IL-15, which maintain or augment CTL activity (55, 56, 57, 58, 59). Both IL-4 and IL-10 mRNA levels increase within the CNS of JHMV-infected mice at the time of viral clearance, although very little IL-4 mRNA can be detected (60). In this regard it is interesting that virus replicates to a higher titer in the CNS of mice rendered deficient in IL-10 by homologous recombination compared with that in control mice (our unpublished observations). Although this may suggest that IL-10 contributes to the CTL activity in the CNS, IL-10 is secreted from CD4- endogenous CNS cells during inflammatory responses (56, 61), and few Th2 type CD4+ T cells are present within the CNS of JHMV-infected mice, as judged by the absent (62) or low (60) levels of IL-4 mRNA detected.
JHMV infection of cells in vitro results in cytopathic effects characterized by giant cell formation. However, in vivo there are few giant cells or apoptotic cells in the absence of an immune response. This suggests that in vivo JHMV infection may be more closely related to infections by noncytopathic viruses, such as LCMV and HIV. Analysis of mice infected with high doses of LCMV suggested that CD4+ T cells may be important in preventing CTL exhaustion due to a high Ag load (2, 3, 22, 23). However, this interpretation is controversial, since other data suggest that CD4+ T cells are required to sustain CTL activity and CTL memory during the high Ag load associated with persistent LCMV infection (18, 21, 22, 63). Whether the requirement for CD4+ T cells is related to their ability to assist in the reduction of viral load during JHMV infection via secretion of antiviral cytokines or the provision of B cell help is not clear. However, CD4+ T cells may also contribute directly to the prevention of CTL exhaustion by the provision of cytokines, such as IL-2. In contrast to JHMV infection of the CNS, the reduction in LCMV-specific CTL activity does not occur until after acute infection, and neither CTL-mediated viral clearance or protection from acute infection is compromised via elimination of CD4+ T cells (18, 21, 22, 29, 63). The JHMV Ag load within the CNS appears sufficient for CD8+ T cells to overcome a requirement for CD4-derived IL-2 during induction (4, 19); however, cytokine secreted by activated CD4+ T cells may be required to maintain the lytic activity or viability of virus-specific CTL within the CNS. In addition to the requirement(s) for CD4+ T cells to secrete cytokines, the ability to inhibit virus replication and mediate protection from JHMV infection may also be attributed to differences in the access of CD8+ T cells to peripheral tissues vs the CNS parenchyma (15).
In the present report the host immune response assisting CTL-mediated clearance of JHMV from the CNS was dependent on CD4+ T cells. The data show that the CTL response induced by JHMV infection in the absence of CD4+ T cells is reduced, but not abrogated, compared with that in normal mice. Therefore, JHMV infection, although restricted to the CNS, is similar to the majority of other viruses examined (1) in that CTL are initially induced in a CD4+ T cell-independent manner. However, in determining the basis for the requirement for CD4+ T cells in the antiviral activity of CTL within the CNS, the data show that activated CTL can access the CNS in the absence of CD4+ T cells. These CTL appear to be nonfunctional, since there is little or no evidence for CTL-mediated virus clearance in CD4+ T cell-depleted mice even following the adoptive transfer of highly activated CTL. This interpretation is consistent with histologic analysis of the CNS of mice depleted of CD4+ T cells, which exhibits a reduction in mononuclear cell infiltration, yet an increase in the number of virus-infected cells. These data suggest that the CNS differs in a fundamental aspect from peripheral organs, which allow ready access to activated CTL that appear to be able to recognize Ag-bearing target cells and clear virus (15).
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. S. Stohlman, University of Southern California, MCH 142, 1333 San Pablo St., Los Angeles, CA 90033.
3 Abbreviations used in this paper: LCMV, lymphocytic choriomeningitis virus; CNS, central nervous system; JHMV, JHM strain of mouse hepatitis virus; N, nucleocapsid; BrdU, bromodeoxyuridine; RCS, rat concanavalin A supernatant; low (superscript), low level; high (superscript), high level.
Received for publication August 20, 1997. Accepted for publication November 19, 1997.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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exerts an antiviral effect in the central nervous system but not in peripheral solid organs. J. Immunol. 150:2316.[Abstract]
4 integrin directs virus-activated CD8+ T cells to sites of infection. J. Immunol. 154:5293.[Abstract]
-chain signaling cytokines regulate activated T cell apoptosis in response to growth factor withdrawal: selective induction of anti-apoptotic (bcl-2, bcl-xL) ut not pro-apoptotic (bax, bcl-xS) gene expression. Eur. J. Immunol. 26:294.[Medline]
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J. M. Gonzalez, C. C. Bergmann, C. Ramakrishna, D. R. Hinton, R. Atkinson, J. Hoskin, W. B. Macklin, and S. A. Stohlman Inhibition of Interferon-{gamma} Signaling in Oligodendroglia Delays Coronavirus Clearance without Altering Demyelination Am. J. Pathol., March 1, 2006; 168(3): 796 - 804. [Abstract] [Full Text] [PDF]
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J. Zhou, N. W. Marten, C. C. Bergmann, W. B. Macklin, D. R. Hinton, and S. A. Stohlman Expression of Matrix Metalloproteinases and Their Tissue Inhibitor during Viral Encephalitis J. Virol., April 15, 2005; 79(8): 4764 - 4773. [Abstract] [Full Text] [PDF]
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K. C. MacNamara, M. M. Chua, P. T. Nelson, H. Shen, and S. R. Weiss Increased Epitope-Specific CD8+ T Cells Prevent Murine Coronavirus Spread to the Spinal Cord and Subsequent Demyelination J. Virol., March 15, 2005; 79(6): 3370 - 3381. [Abstract] [Full Text] [PDF]
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C. Ramakrishna, S. A. Stohlman, R. A. Atkinson, D. R. Hinton, and C. C. Bergmann Differential Regulation of Primary and Secondary CD8+ T Cells in the Central Nervous System J. Immunol., November 15, 2004; 173(10): 6265 - 6273. [Abstract] [Full Text] [PDF]
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 U. Fassnacht, A. Ackermann, P. Staeheli, and J. Hausmann Immunization with dendritic cells can break immunological ignorance toward a persisting virus in the central nervous system and induce partial protection against intracerebral viral challenge J. Gen. Virol., August 1, 2004; 85(8): 2379 - 2387. [Abstract] [Full Text] [PDF]
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W. G. Glass, M. J. Hickey, J. L. Hardison, M. T. Liu, J. E. Manning, and T. E. Lane Antibody Targeting of the CC Chemokine Ligand 5 Results in Diminished Leukocyte Infiltration into the Central Nervous System and Reduced Neurologic Disease in a Viral Model of Multiple Sclerosis J. Immunol., April 1, 2004; 172(7): 4018 - 4025. [Abstract] [Full Text] [PDF]
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C. C. Bergmann, B. Parra, D. R. Hinton, C. Ramakrishna, K. C. Dowdell, and S. A. Stohlman Perforin and Gamma Interferon-Mediated Control of Coronavirus Central Nervous System Infection by CD8 T Cells in the Absence of CD4 T Cells J. Virol., February 15, 2004; 78(4): 1739 - 1750. [Abstract] [Full Text] [PDF]
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M. M. Chua, K. C. MacNamara, L. San Mateo, H. Shen, and S. R. Weiss Effects of an Epitope-Specific CD8+ T-Cell Response on Murine Coronavirus Central Nervous System Disease: Protection from Virus Replication and Antigen Spread and Selection of Epitope Escape Mutants J. Virol., February 1, 2004; 78(3): 1150 - 1159. [Abstract] [Full Text] [PDF]
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R. S. Goldszmid, J. Idoyaga, A. I. Bravo, R. Steinman, J. Mordoh, and R. Wainstok Dendritic Cells Charged with Apoptotic Tumor Cells Induce Long-Lived Protective CD4+ and CD8+ T Cell Immunity against B16 Melanoma J. Immunol., December 1, 2003; 171(11): 5940 - 5947. [Abstract] [Full Text] [PDF]
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C. Ramakrishna, C. C. Bergmann, R. Atkinson, and S. A. Stohlman Control of Central Nervous System Viral Persistence by Neutralizing Antibody J. Virol., April 15, 2003; 77(8): 4670 - 4678. [Abstract] [Full Text] [PDF]
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D. Stober, I. Jomantaite, R. Schirmbeck, and J. Reimann NKT Cells Provide Help for Dendritic Cell-Dependent Priming of MHC Class I-Restricted CD8+ T Cells In Vivo J. Immunol., March 1, 2003; 170(5): 2540 - 2548. [Abstract] [Full Text] [PDF]
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A. A. Dandekar and S. Perlman Virus-Induced Demyelination in Nude Mice Is Mediated by {gamma}{delta} T Cells Am. J. Pathol., October 1, 2002; 161(4): 1255 - 1263. [Abstract] [Full Text] [PDF]
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C. E. Patterson, D. M. P. Lawrence, L. A. Echols, and G. F. Rall Immune-Mediated Protection from Measles Virus-Induced Central Nervous System Disease Is Noncytolytic and Gamma Interferon Dependent J. Virol., March 27, 2002; 76(9): 4497 - 4506. [Abstract] [Full Text] [PDF]
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S.-I. Tschen, C. C. Bergmann, C. Ramakrishna, S. Morales, R. Atkinson, and S. A. Stohlman Recruitment Kinetics and Composition of Antibody-Secreting Cells Within the Central Nervous System Following Viral Encephalomyelitis J. Immunol., March 15, 2002; 168(6): 2922 - 2929. [Abstract] [Full Text] [PDF]
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C. Ramakrishna, S. A. Stohlman, R. D. Atkinson, M. J. Shlomchik, and C. C. Bergmann Mechanisms of Central Nervous System Viral Persistence: the Critical Role of Antibody and B Cells J. Immunol., February 1, 2002; 168(3): 1204 - 1211. [Abstract] [Full Text] [PDF]
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B. Wang, C. C. Norbury, R. Greenwood, J. R. Bennink, J. W. Yewdell, and J. A. Frelinger Multiple Paths for Activation of Naive CD8+ T Cells: CD4-Independent Help J. Immunol., August 1, 2001; 167(3): 1283 - 1289. [Abstract] [Full Text] [PDF]
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C. C. Bergmann, C. Ramakrishna, M. Kornacki, and S. A. Stohlman Impaired T Cell Immunity in B Cell-Deficient Mice Following Viral Central Nervous System Infection J. Immunol., August 1, 2001; 167(3): 1575 - 1583. [Abstract] [Full Text] [PDF]
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J. S. Haring, L. L. Pewe, and S. Perlman High-Magnitude, Virus-Specific CD4 T-Cell Response in the Central Nervous System of Coronavirus-Infected Mice J. Virol., March 15, 2001; 75(6): 3043 - 3047. [Abstract] [Full Text]
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K. H. Edelmann and C. B. Wilson Role of CD28/CD80-86 and CD40/CD154 Costimulatory Interactions in Host Defense to Primary Herpes Simplex Virus Infection J. Virol., January 15, 2001; 75(2): 612 - 621. [Abstract] [Full Text]
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G. Weidinger, S. Czub, C. Neumeister, P. Harriott, V. ter Meulen, and S. Niewiesk Role of CD4+ and CD8+ T cells in the prevention of measles virus-induced encephalitis in mice J. Gen. Virol., November 1, 2000; 81(11): 2707 - 2713. [Abstract] [Full Text]
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P. R. Walker, T. Calzascia, V. Schnuriger, N. Scamuffa, P. Saas, N. de Tribolet, and P.-Y. Dietrich The Brain Parenchyma Is Permissive for Full Antitumor CTL Effector Function, Even in the Absence of CD4 T Cells J. Immunol., September 15, 2000; 165(6): 3128 - 3135. [Abstract] [Full Text] [PDF]
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N. W. Marten, S. A. Stohlman, and C. C. Bergmann Role of Viral Persistence in Retaining CD8+ T Cells within the Central Nervous System J. Virol., September 1, 2000; 74(17): 7903 - 7910. [Abstract] [Full Text]
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R. G. van der Most, K. Murali-Krishna, R. Ahmed, and J. H. Strauss Chimeric Yellow Fever/Dengue Virus as a Candidate Dengue Vaccine: Quantitation of the Dengue Virus-Specific CD8 T-Cell Response J. Virol., September 1, 2000; 74(17): 8094 - 8101. [Abstract] [Full Text]
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G. F. Wu, A. A. Dandekar, L. Pewe, and S. Perlman CD4 and CD8 T Cells Have Redundant But Not Identical Roles in Virus-Induced Demyelination J. Immunol., August 15, 2000; 165(4): 2278 - 2286. [Abstract] [Full Text] [PDF]
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N. W. Marten, S. A. Stohlman, R. D. Atkinson, D. R. Hinton, J. O. Fleming, and C. C. Bergmann Contributions of CD8+ T Cells and Viral Spread to Demyelinating Disease J. Immunol., April 15, 2000; 164(8): 4080 - 4088. [Abstract] [Full Text] [PDF]
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C. C. Bergmann, J. D. Altman, D. Hinton, and S. A. Stohlman Inverted Immunodominance and Impaired Cytolytic Function of CD8+ T Cells During Viral Persistence in the Central Nervous System J. Immunol., September 15, 1999; 163(6): 3379 - 3387. [Abstract] [Full Text] [PDF]
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 G. Perrin, V. Schnuriger, A.-L. Quiquerez, P. Saas, C. Pannetier, N. de Tribolet, J.-M. Tiercy, J.-P. Aubry, P.-Y. Dietrich, and P. R. Walker Astrocytoma infiltrating lymphocytes include major T cell clonal expansions confined to the CD8 subset Int. Immunol., August 1, 1999; 11(8): 1337 - 1350. [Abstract] [Full Text] [PDF]
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M. T. Lin, D. R. Hinton, N. W. Marten, C. C. Bergmann, and S. A. Stohlman Antibody Prevents Virus Reactivation Within the Central Nervous System J. Immunol., June 15, 1999; 162(12): 7358 - 7368. [Abstract] [Full Text] [PDF]
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A.-X. Holterman, K. Rogers, K. Edelmann, D. M. Koelle, L. Corey, and C. B. Wilson An Important Role for Major Histocompatibility Complex Class I-Restricted T Cells, and a Limited Role for Gamma Interferon, in Protection of Mice against Lethal Herpes Simplex Virus Infection J. Virol., March 1, 1999; 73(3): 2058 - 2063. [Abstract] [Full Text]
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 D. Schluter, E. Domann, C. Buck, T. Hain, H. Hof, T. Chakraborty, and M. Deckert-Schluter Phosphatidylcholine-Specific Phospholipase C from Listeria monocytogenes Is an Important Virulence Factor in Murine Cerebral Listeriosis Infect. Immun., December 1, 1998; 66(12): 5930 - 5938. [Abstract] [Full Text] [PDF]
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S. Rassnick, L. W. Enquist, A. F. Sved, and J. P. Card Pseudorabies Virus-Induced Leukocyte Trafficking into the Rat Central Nervous System J. Virol., November 1, 1998; 72(11): 9181 - 9191. [Abstract] [Full Text] [PDF]
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