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Respiratory Medicine, National Heart and Lung Institute, Imperial College of Science, Technology and Medicine, London, United Kingdom
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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rendered C57BL/6, but
not BALB.K mice, susceptible to eosinophilia. Analysis of B10
recombinant mice showed that the Dd allele (in B10.A(5R)
mice) prevented CD8+ T cell accumulation in the lung,
resulting in intense lung eosinophilia. However, the Db
allele (in B10.A(2R) and B10.A(4R) mice) supported CD8+ T
cell expansion and prevented eosinophilia. Intracellular cytokine
staining showed that lung eosinophilia correlated with reduced IFN-
and increased IL-10 expression in lung T cells. These results are
compatible with the unifying model that Th2 cells mediate the disease
but can be inhibited by CD8+ T cells secreting IFN-. Our
findings have important implications for the development of protective,
nonpathogenic vaccines for RSV disease. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The BALB/c mouse model of RSV infection reproduces some important features of the human disease and has provided many insights into possible protective and pathogenic immune processes in man. Protective Ab responses are directed primarily against the fusion (F) and attachment (G) glycoproteins, both of which are expressed on the surface of the virion. Vaccination with F expressed by recombinant vaccinia virus (rVV) generates Ab, cytotoxic T cells, and CD4+ Th1 cells after virus challenge (1, 2, 3, 4), whereas the glycoprotein G of RSV (rVV-G) leads to Ab production and CD4+ Th2 cells, but no detectable cytotoxic T cells (5, 6, 7, 8, 9). The patterns of lung pathology after virus challenge are also distinctive, in that F-primed mice develop augmented lung disease characterized by lung hemorrhage and neutrophilia, whereas G-primed mice develop pulmonary eosinophilia (10, 11).
Pulmonary eosinophilia was also found in the lungs and
peripheral blood of some children vaccinated with formalin inactivated
RSV (see, for example, 12 . To improve our understanding of
vaccine augmentation, we and others have performed extensive studies of
the immune determinants of RSV-induced lung eosinophilia in BALB/c mice
sensitized to G. In drawing general conclusions about the relevance of
this response to disease in diverse hosts, mapping genetic responses in
inbred mice appears vital. It is well established that CD4+
T cells are instrumental in causing eosinophilic disease augmentation,
and that CD8+ T cells making IFN- may be crucial
regulators of this response (5, 13). MHC gene effects are therefore
likely to play a critical role in the genetic control of augmented RSV
disease. Associations between MHC and susceptibility to infection have
been reported (14, 15, 16, 17, 18) and indicate the difficulties encountered when
trying to design vaccines for outbred human populations. Different
inbred strains of mice have provided an insight into the influence of
MHC haplotype on the outcome of infection in many infectious disease
models. However, there has been no systematic study of the genetic
influences on the induction of vaccine-augmented pathology during RSV
infection.
In the present studies, we sought to determine the immunologic
mechanisms and genetic influences on RSV-induced lung eosinophilia
using 15 inbred and 7 first generation (F1)
crossbred mice. Our results indicate that eosinophilic lung
disease requires CD4+ T cell recognition, and that, in
mouse strains susceptible to eosinophilia, CD8+ T cells
secreting IFN- play an important regulatory role. The results
obtained using different inbred and recombinant mouse strains may
explain the variability in the response to RSV infection in humans and
give insights into mechanisms that must be considered in designing
vaccines for use in outbred human populations.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Recombinant vaccinia expressing rVV-G and the A2 strain of RSV were grown in HEp-2 cells, snap frozen in liquid nitrogen, and assayed for infectivity. Vaccinia expressing ß-galactosidase (rVV-ß-gal) or HEp-2 cells were used as respective controls. All virus and control Ag preparations were shown to be free of mycoplasma contamination by DNA hybridization (Gen-Probe, San Diego, CA).
Mice
Eight- to 10-wk-old female mice were purchased from Harlan Olac (Bicester, U.K.) and kept in pathogen-free conditions. The following strains of mice were used: BALB/c, DBA/2n, B10.D2 (all H-2d); BALB.B, 129, C57BL/6, C57BL/10 (all H-2b); BALB.K, B10.BR, CBA/Ca, CBA/J, C3H/HeN (all H-2k); and B10.A (2R), (4R) and 5(R) (H-2 h2, h4, and i2, respectively). First generation crosses were derived between the following mouse strains: C3H x DBA/2n, B10.BR x B10.D2, BALB.K x B10.D2, BALB.K x BALB/c, B10.BR x BALB/c (all H-2dxk), and C57BL/6 x BALB/c, C57BL/6 x B10.D2 (both H-2b/d) (for a review of these mouse strains see 19 . On day 0, groups of 4–5 mice were scarified on the rump with either 3 x 106 plaque-forming units of rVV-G or rVV-ß-gal in a final volume of 10 µl as previously described (20). On day 14, mice were challenged intranasally with 50 µl of RSV stock (equivalent to 106 plaque-forming units/mouse) or HEp-2 control Ag. All animal experimentation was performed under the guidelines set out by the Home Office.
Determination of vaccinia virus replication in different strains of mice
The skin from individual mice, sensitized with rVV-G, was removed at different time points after scarification and placed in serum-free medium (RPMI 1640). The skin and s.c. tissue were mechanically disrupted, and 106 cells were pelleted and sonicated for 1 min in a water bath sonicator. The supernatant was assayed for vaccinia virus using a standard HEp-2 cell plaque assay.
Determination of respiratory syncytial virus titers in lung tissue
Lungs were removed from four G-primed mice 4 and 7 days after RSV challenge and homogenized. Clarified supernatant was added to a HEp-2 monolayer and virus titers were determined as described (21).
Recovery of lung cells
After 7 days, mice were injected i.p. with 3 mg/mouse pentobarbitone and exanguinated via the femoral vessels. Bronchoalveolar lavage (BAL) was performed as described previously (20). Two hundred microliters of lavage fluid from each mouse was stored in individual Eppendorf tubes for subsequent cytospin preparations. Samples from individual mice were examined by flow cytometry for expression of surface markers and intracellular cytokine expression. All samples were kept in ice cold RPMI 1640 containing 10% FCS, 2 mM/ml L-glutamine, penicillin (50 U/ml), and streptomycin (50 µg/ml) (R10F). Mediastinal lymph nodes were removed from individual mice in each experimental group and placed in ice cold R10F. All samples were washed, counted, and resuspended at a concentration of 106 cells/ml.
Cytocentrifuge preparations
Lavage cells from individual mice were cytocentrifuged onto glass slides at 700 rpm for 5 min. Slides were air-dried for 30 min and stained with Wright/Giemsa (Sigma Immunochemicals, Dorset, U.K.) according to the manufacturer’s instructions. At least 300 cells were counted in three separate fields for each preparation. Eosinophils were identified by their distinct eosinophilic granules on light microscopy (allowing them to be distinguished form other granulocytes) and, additionally, by size and granularity on flow cytometry.
Flow cytometric analysis of intracellular and surface Ags
Intracellular IFN- (AN18-FITC), IL-4 (11B11-PE), IL-5
(TRFK5-PE), and IL-10 (JES5–16E3 PE) were assessed in CD4+
(H129.19-QR) or CD8+ (15–6.7-QR) T cells by flow cytometry
as described previously (22). Unstimulated cells from individual mice
were also stained with Abs to CD4 (QR), CD8 (FITC), and CD45RB (PE).
Samples were analyzed on a Coulter (Luton, U.K.) EPICS Elite flow
cytometer, collecting data on at least 30,000 gated lymphocytes from
each sample.
Determination of G-specific Ab by ELISA
G-specific Ab was assessed by ELISA as described previously (20). ELISA Ag was prepared by infecting HEp-2 cells with either rVV-G or rVV-ß-gal. When significant cytopathic effect was observed, the infected cells were harvested, centrifuged, resuspended in 500 µl, and then subjected to 2 min of sonication in a sonicator (Ultrawave, Cardiff, U.K.). Aliquots (10 µl) were stored at -20°C until required. Briefly, microtiter plates were coated overnight with 100 µl of either rVV-G or control rVV-ß-gal Ag. After blocking with 2% normal rabbit serum for 2 h, dilutions of test sample were added for another hour at room temperature. Bound Ab was detected using peroxidase-conjugated rabbit anti-mouse Ig and the o-phenylenediamine substrate. Color changes were quantified in an ELISA plate reader at 490 nm. This method to detect G-specific Ab was standardized using G-specific mAbs (kindly provided by Dr. Jose Melero, Madrid, Spain). The amount of G-specific Ab was determined by subtracting the optical density obtained by incubating serum on rVV-ß-gal-coated plates from the same sample incubated on rVV-G coated plates.
Depletion of T cell subsets
Mice were treated with an i.p. injection of 200 µg/mouse/day
of depleting Abs to CD4 (YTS191.1 + YTA3.1) or CD8 (YTS169.4 +
YTS156.7) Abs from day 8 to 18 after scarification with rVV-G. On day
14, mice were challenged intranasally with RSV, and tissues were
sampled on day 21. Depletion of IFN- was performed by i.p. injection
of rat anti-mouse IFN- (XMG 1.2) or control Ab, starting 2 days
before RSV challenge and daily for 5 days, repeated on days 4, 7, and
10 of infection.
Statistical analysis
The significance of differences was assessed by Student’s t test.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The development of pulmonary eosinophilia in G-primed BALB/c mice
after RSV challenge has been extensively documented (for a review, see
23 . However, when other inbred strains of mice were treated in a
similar manner, distinct patterns of pathology were observed (Fig. 1
). Similar to BALB/c mice, other strains
with MHC H-2d haplotype developed an eosinophilic response
to G priming after intranasal RSV challenge. Though variable
percentages of eosinophils were noted in different H-2d
strains, there was little difference in the absolute number of
eosinophils among these strains. However, no eosinophils were recovered
from the BAL of mouse strains with an H-2k haplotype,
despite developing an enhanced infiltrate compared with control-primed
mice after challenge. Mice with a MHC H-2b haplotype showed
various responses, some strains developing eosinophilia and others not.
The lack of eosinophilia in two mouse strains with H-2b
haplotype was associated with a C57BL background (C57BL/6 and
C57BL/10). Eosinophilia in sensitive mouse strains was specifically
associated with a secondary immune response to RSV, as no mouse strain
tested developed eosinophilia after RSV challenge when previously
sensitized with the control construct (rVV-ß-gal, data not shown).
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To examine whether the differential induction of eosinophilia
might result from altered replication of vaccinia virus or RSV, we
measured virus replication by plaque assay using lung homogenates or
skin from different inbred strains of mice. All mice cleared vaccinia
virus by day 14 after scarification. In addition, in all of the mouse
strains tested, the inguinal lymph nodes (draining the site of
scarification) expanded at a similar rate, confirming that efficient
priming had occurred (data not shown). As in previous studies, higher
titers of RSV were recovered from mice undergoing a primary RSV
infection (ß-gal-primed mice) than from those pre-sensitized with
rVV-G. RSV replication was similar in mouse strains previously
sensitized to G, regardless of whether they developed eosinophilia or
not (p > 0.05 for day 4 and day 7,
respectively, by Student’s t test; Fig. 2).
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The differential induction of eosinophils in the various inbred
mouse strains may have reflected differences in the extent, rather than
the type, of immune response to the virus. Therefore, total cell
recovery, lymphocytic infiltrate, and the proportion of CD4:CD8 T cells
from the BAL of mice were examined during primary and secondary
infections. There were no significant differences in total cells
(p > 0.05) or lymphocytic responses
(p > 0.05) between mouse strains of different
MHC haplotype (data not shown). Pre-sensitization with the G protein
resulted in enhanced cell accumulation in H-2d and
H-2b mice after viral challenge compared with mice
undergoing a primary RSV infection, indicating that all mice developed
an immune response to vaccination with rVV-G (p
< 0.01 comparing primary and secondary cell accumulation for each
mouse strain). The increase in cell accumulation after G protein
sensitization also occurred in all H-2k mouse strains but
was less marked (p 
0.039 comparing primary and
secondary cell recruitment).
As CD4+ T cells are necessary for eosinophil recruitment in
this model, we then examined the proportion of CD4+ and
CD8+ T cells in BAL samples by flow cytometry. No
significant difference was observed in CD4+ and
CD8+ T cell responses between mice of different MHC
haplotype during primary RSV infection (p >
0.05; Fig. 3). However, during secondary
infection after G protein priming, mice that developed lung
eosinophilia had either enhanced CD4+ T cells, reduced
CD8+ T cells, or both, compared with mice of the same
strain undergoing a primary immune response (Fig. 3). In addition,
CD8+ T cells were significantly decreased in mice that
developed eosinophilia compared with other G-vaccinated mice that did
not (p < 0.01). In H-2k mice,
which did not develop eosinophilia, there was no significant difference
in the proportion of T cell subsets present during primary and
secondary infection in three strains (BALB.K, CBA/J, and B10.BR;
p > 0.05). However, CBA/Ca and C3H mice did show
enhanced CD4+ T cell recruitment after G priming (Fig. 3).
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G-specific Ab production
CD4+ T cells are required for induction of
RSV-specific Ab, whereas CD8+ T cells are inhibitory. The
presence of RSV-specific Ab therefore reflects the relative balance of
these two T cell subsets. All G protein-sensitized mice generated
specific Ab after RSV challenge (Fig. 4),
which was higher than that obtained for the same mouse strain
undergoing a primary immune response to RSV. For clarity only, Ab
generated to G priming followed by RSV challenge is shown. All mice
displayed G-specific Ab production. Though two of the H-2d
mice gave higher optical density readings at low serum dilutions, the
end point titer was similar in all mouse strains tested.
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To investigate the possible mechanisms of eosinophil induction or
inhibition we examined intracellular IFN-, IL-4, IL-5, and IL-10
expression in CD4+ and CD8+ T cells by flow
cytometry. Mice that developed eosinophilia to G had significantly
reduced IFN- in both CD4+ and CD8+ BAL T
cells (p < 0.001 for both) (Fig. 5). IFN- expression in
CD4+ and CD8+ mediastinal lymph node T cells
was similar in all groups (not shown). No significant IL-4 or IL-5 was
detected in any T cell population from the lung or any other site.
IL-10, however, was detected in CD4+ and CD8+ T
cells in the BAL, more so in mouse strains that developed eosinophilia
than in those that did not (Fig. 6,
p < 0.05 by Student’s t test). The
majority of IL-10 expression was within cells that also co-expressed
IFN-.
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To determine the influence of background genes and MHC haplotype
on eosinophil induction by the G protein of RSV, first generation
crosses of mice that developed eosinophilia were bred with strains of
mice that do not. All H-2d x H-2k, but no
H-2d x H-2b, first generation mice tested
developed eosinophilia (Fig. 7).
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Mice resistant to eosinophil induction by G protein priming had
higher levels of IFN- in CD8+ T cells compared with
sensitive strains. We therefore depleted CD8+ T cells using
neutralizing Abs before and during challenge. Removal of
CD8+ T cells from C57BL/6, but not BALB.K, mice resulted in
lung eosinophilia (Fig. 8). Similar
observations were observed by depletion of IFN- (data not shown).
Eosinophil induction in C57BL/6 mice was not due to the Ab treatment,
as depletion of both CD4+ and CD8+ T cells
abolished BAL eosinophilia. In addition, we have shown previously that
C57BL/6 mice with CD8
-/-,
ß2-microglobulin-/-, and
TAP1-/- gene deletions similarly develop eosinophilia to
the G protein (13). The present results suggest that elements
responsible for resistance to eosinophilia are different in mice of
H-2b and H-2k haplotype.
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Immune responses to the G protein in recombinant mice
As CD8+ T cells appeared an important regulator of
eosinophilic pathology, we examined the effect of G priming and RSV
challenge in mice with a B10 background but expressing different MHC
alleles. G-primed B10.A(5R) mice expressing H-2Dd developed
lung eosinophilia after intranasal virus challenge, whereas B10.A(4R)
and B10.A(2R) mice, which both express H-2Db, did not (Fig. 9, top). CD4+ T
cells were enhanced (Fig. 9, middle), and CD8+ T
cells effectively abrogated (Fig. 9, bottom) in B10.A(5R)
mice. In addition, CD4+ T cells from these mice were highly
activated based on the increased loss of CD45RB staining compared with
the same cells from B10.A(2R) and (4R) mice (Fig. 10).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To summarize current knowledge of the BALB/c mouse model of RSV
infection, induction of lung eosinophilia requires priming of
CD4+ T cells that make IL-4 and IL-5. Depletion of these
cells with anti-CD4 mAbs prevents eosinophilia. Eosinophilia
can be transferred to naïve recipients with RSV specific
CD4+ T cells that make Th2 cytokines in vitro. The effects
of these Th2 cells can be antagonized by virus-specific
CD8+ T cells, which make IFN- (5, 7, 13), by
administration of IL-12 during vaccination (20), or by the removal of
immunodominant CD4+ T cell epitope(s) from the G protein
(24). According to this model, G protein primes for eosinophilia
because it induces only CD4+ T cells. Its failure to prime
CD8+ T cells (5, 6) is responsible for allowing the
CD4+ T cells to default to a Th2 pattern of cytokine
production, which leads to eosinophilia.
Our initial studies concentrated on comparison of C57BL/6 and BALB/c mice. These are the most common inbred mouse strains used to study infectious agents and often show contrasting patterns of immunity and disease. For example, during Leishmania major infection, BALB/c mice show a strong CD4+ Th2 cell response and are incapable of clearing infection, whereas C57BL/6, B10.D2, and C3H mouse strains that resolve infection show strong Th1 responses (25, 26). By contrast, mouse strains that show a strong Th2 response during infection with Trichuris muris are resistant to infection, whereas those that mount a Th1 response fail to expel these worms (27, 28, 29). It is said that C57BL/6, B10.D2, and C3H mice tend to develop Th1 responses, but that a BALB background results in Th2 response to many Ags (30, 31). However, previously published studies have generally been confined to small numbers of inbred and congenic mice, limiting the interpretation that can be placed on the results presented in our study.
Our results with H-2d mice are compatible with the
hypothesis that eosinophilia is simply a result of unbalanced
CD4+ T cell induction. Eosinophils were observed in G
protein primed DBA/2n and B10.D2 (DBA/2n MHC on a BL/10 background)
mice after RSV infection. In these mice, it seems that H-2d
presents appropriate epitopes to CD4+ T cells, but that
CD8+ T cell responses are not induced. Failure of IFN-
production by CD8+ T cells allows Th2 cells to develop, and
lung eosinophilia therefore ensues. The results in H-2b
mice were more complex. C57BL/6 and C57BL/10 mice were resistant to
G-induced eosinophilia, but on a BALB background (BALB.B)
H-2b renders mice susceptible. In BALB.B mice,
CD4+ T cell depletion prevents lung eosinophilia, showing
that H-2b can induce a pathogenic CD4+ T cell
response. We suggest that the variation in eosinophilia in
H-2b mice must be due either to background genes or to
variations in the levels of T cell cytokines in different strains.
Indeed, CD4+ and CD8+ T cells produced high
levels of IFN- and lower levels of IL-10 in C57BL/6 and C57BL/10
mice compared with BALB.B and 129 mice primed with the G protein.
Previous studies have classified C57BL/6 mice as high IFN-/ß
producers (If-1h), whereas BALB/c mice are low producers
(If-1l). This effect has been demonstrated during Newcastle
disease virus infection where C57BL/6 mice show 10- to 15-fold higher
circulating levels of IFN than BALB/c mice (32). Although innate
immunity (such as IFN-/ß production) may vary with background
genes, there is insufficient evidence to judge whether the variations
in eosinophilia in MHC H-2b mouse strains could be
explained by such factors.
C57BL/6 and 129 mice also differ in Mls alleles that may influence immune responses to G. 129 mice express the Mls2a allele similar to BALB/c mice, whereas C57BL/6 mice express Mls2b (33, 34). Such differences, and the distribution of mouse mammary tumor virus variants, may affect the TCR Vß repertoires and subsequent immunity to the G protein.
Using recombinant B10 mouse strains, we were able to show that expression of Db protected mice from eosinophilia after G protein priming, whereas expression of Dd did not. In B10 mice with H-2 Dd, BAL CD8+ T cells were dramatically reduced, and CD4+ T cell recruitment and activation (assessed by CD45RB expression) were increased. Such mice developed extensive lung eosinophilia when primed with the G protein. This result underscores the premise that CD8+ T cell activation has a critical influence on CD4+ T cell activation and subsequent eosinophilia.
The absence of eosinophilia in G-primed H-2k mouse
strains is not explained by background genes, because simple
substitution of H-2k onto the BALB background (BALB.K) does
not induce lung eosinophilia. In contrast to the effects in C57BL/6
mice, removal of CD8+ T cells using depleting Ab did not
allow eosinophilia to develop in BALB.K mice. We have not yet
determined whether CD8+ T cell depletion induces
eosinophilia in other mouse strains of H-2k haplotype. Our
previous work showed that CD4+ T cells are necessary for
eosinophil induction (13), and, therefore, an alternative explanation
for the lack of eosinophilia is that H-2k mice fail to
generate CD4+ T cell responses to G protein. Our finding
that three out of the five H-2k mouse strains did not
display enhanced CD4+ T cell recruitment, which we have
observed to be a reliable indicator of secondary CD4+ T
cell responses, supports this possibility. However, we cannot rule out
the possibility that T cells secreting type 1 rather than type 2
cytokines are induced. Although CD4+ T cell recruitment was
less prominent in H-2k than in H-2d mice, those
T cells that were present in the lungs of H-2k mice
expressed higher levels of intracellular IFN-.
First generation crosses between mice with MHC H-2d and H-2k were susceptible to G-induced lung eosinophilia. Whether H-2k mice recognize the G protein or not, the H-2d response (which we know recognizes a CD4+ T cell epitope in G) is dominant. Such differential induction of eosinophilia in H-2d (BALB/c), but not H-2k (B10.BR), mice has previously been shown during sensitization with Dermatophagoides farinae (35). In this study, the only noticeable difference between the two strains was the lack of an early type hypersensitivity response in B10.BR mice compared with BALB/c. It was surprising that F1 crosses between H-2d mice and C57BL/6 (H-2b) did not result in lung eosinophilia. We know that both of these mouse strains induce a CD4+ T cell response and that H-2d cannot support CD8+ T cell response (13). Therefore, this first generation cross includes CD4+ T cell responses via both MHC haplotypes but CD8+ T cell responses from only the H-2b MHC. As depletion of CD8+ T cells renders C57BL/6 mice susceptible to G-induced eosinophilia, we believe that induction of such cells is responsible for the result obtained with the first generation cross. Other possible influences in this F1 include competition between d and b alleles for expression on the cell surface, presentation of an alternative epitope that does not induce eosinophils, tolerization to eosinophilic epitope(s) in G, and processing and presentation alterations.
Though the differential induction of CD4+ and CD8+ T cells explains most of the results presented in this study, numerous parameters have been highlighted in other mouse models of infectious diseases. Airway responsiveness (36, 37), the number of T cell epitopes recognized (38), the avidity or affinity of ligand:T cell interaction (39, 40, 41, 42), eosinophil precursor frequency (27, 43), and the dose of Ag (44, 45) are all consistent with the mouse strain profiles presented in our study.
The lack of IL-4 and IL-5 production by T cells ex vivo using
intracellular cytokine staining is interesting. We and others have
previously observed Th2 cytokine production in T cells from G-primed
mice using either RT-PCR or ELISA (5, 46, 47). Th2 cytokine mRNA is
only transiently expressed very early after RSV challenge in G-primed
mice, and the source of this mRNA is undetermined (46). Therefore, we
used intracellular cytokine staining in this study. We have previously
shown that IFN- and IL-10 accumulation detected by intracellular
staining correlates well with cytokine mRNA (22). We believe that the
results presented in this paper are an accurate reflection of events
occurring in vivo. Though we have detected IL-4 and IL-5 by cytokine
staining (20), future studies would benefit from an analysis of
cytokines by several complementary methods.
The differential induction of eosinophilia does not reflect variable replication of either the vaccinia virus construct or RSV in these mice. Though the study by Prince et al. (48) shows that different strains of 3-day-old mice vary in the titer of RSV recoverable from the lungs, we did not observe differences of this type in 8- to 10-wk-old mice. Furthermore, the results do not reflect differences in the severity of the inflammatory reaction, since total cell recovery and lymphocytic infiltrate did not significantly differ between mouse strains. What did correlate with eosinophilia was the level of CD4+ and CD8+ T cell induction and the level of cytokines expressed by these cells.
Since the G protein is a surface protein and a target of neutralizing Ab, its inclusion in a vaccine would be logical. The present study indicates that a safe vaccine might be one that induces a strong CD8+ T cell response. Despite the possible influence of non-MHC genes in some mouse strains, the effects seen in inbred, crossbred, and recombinant mice and the effect of T cell depletions shows that MHC is the primary determinant of exuberant T cell responses and eosinophilia during RSV challenge. Our results are consistent with a simple model of T cell induced disease, which may help explain the variability in response to vaccination and RSV infection in humans.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Tracy Hussell, Respiratory Medicine, National Heart and Lung Institute, Imperial College of Science, Technology and Medicine, St Mary’s Hospital, Norfolk Place, London W2 1PG, U.K. E-mail address:
3 Abbreviations used in this paper: RSV, respiratory syncytial virus; F, fusion glycoprotein; G, attachment protein; rVV, recombinant vaccinia virus; ß-gal, ß-galactosidase; BAL, bronchoalveolar lavage.
Received for publication May 13, 1998. Accepted for publication July 31, 1998.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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or interleukin 4 during the resolution or progression of murine leishmaniasis. J. Exp. Med. 169:59./ß-transgenic model. J. Exp. Med. 182:1579. production by local T cells in human respiratory syncytial virus-induced eosinophilic lung disease. J. Gen. Virol. 79:1751.[Abstract]
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G. A. Melendi, S. J. Hoffman, R. A. Karron, P. M. Irusta, F. R. Laham, A. Humbles, B. Schofield, C.-H. Pan, R. Rabold, B. Thumar, et al. C5 Modulates Airway Hyperreactivity and Pulmonary Eosinophilia during Enhanced Respiratory Syncytial Virus Disease by Decreasing C3a Receptor Expression J. Virol., January 15, 2007; 81(2): 991 - 999. [Abstract] [Full Text] [PDF]
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F. J. Culley, A. M. J. Pennycook, J. S. Tregoning, T. Hussell, and P. J. M. Openshaw Differential Chemokine Expression following Respiratory Virus Infection Reflects Th1- or Th2-Biased Immunopathology J. Virol., May 1, 2006; 80(9): 4521 - 4527. [Abstract] [Full Text] [PDF]
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B. D. Rudd, J. J. Smit, R. A. Flavell, L. Alexopoulou, M. A. Schaller, A. Gruber, A. A. Berlin, and N. W. Lukacs Deletion of TLR3 Alters the Pulmonary Immune Environment and Mucus Production during Respiratory Syncytial Virus Infection J. Immunol., February 1, 2006; 176(3): 1937 - 1942. [Abstract] [Full Text] [PDF]
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 R. A. Tripp, C. Oshansky, and R. Alvarez Cytokines and Respiratory Syncytial Virus Infection Proceedings of the ATS, August 1, 2005; 2(2): 147 - 149. [Abstract] [Full Text] [PDF]
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 P. J. M. Openshaw and J. S. Tregoning Immune Responses and Disease Enhancement during Respiratory Syncytial Virus Infection Clin. Microbiol. Rev., July 1, 2005; 18(3): 541 - 555. [Abstract] [Full Text] [PDF]
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T. R. Johnson, S. E. Mertz, N. Gitiban, S. Hammond, R. LeGallo, R. K. Durbin, and J. E. Durbin Role for Innate IFNs in Determining Respiratory Syncytial Virus Immunopathology J. Immunol., June 1, 2005; 174(11): 7234 - 7241. [Abstract] [Full Text] [PDF]
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B. D. Rudd, E. Burstein, C. S. Duckett, X. Li, and N. W. Lukacs Differential Role for TLR3 in Respiratory Syncytial Virus-Induced Chemokine Expression J. Virol., March 15, 2005; 79(6): 3350 - 3357. [Abstract] [Full Text] [PDF]
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S. P. Matthews, J. S. Tregoning, A. J. Coyle, T. Hussell, and P. J. M. Openshaw Role of CCL11 in Eosinophilic Lung Disease during Respiratory Syncytial Virus Infection J. Virol., February 15, 2005; 79(4): 2050 - 2057. [Abstract] [Full Text] [PDF]
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T. R. Johnson, S. M. Varga, T. J. Braciale, and B. S. Graham V{beta}14+ T Cells Mediate the Vaccine-Enhanced Disease Induced by Immunization with Respiratory Syncytial Virus (RSV) G Glycoprotein but Not with Formalin-Inactivated RSV J. Virol., August 15, 2004; 78(16): 8753 - 8760. [Abstract] [Full Text] [PDF]
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T. R. Johnson, M. N. Teng, P. L. Collins, and B. S. Graham Respiratory Syncytial Virus (RSV) G Glycoprotein Is Not Necessary for Vaccine-Enhanced Disease Induced by Immunization with Formalin-Inactivated RSV J. Virol., June 1, 2004; 78(11): 6024 - 6032. [Abstract] [Full Text] [PDF]
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T. R. Johnson, R. A. Parker, J. E. Johnson, and B. S. Graham IL-13 Is Sufficient for Respiratory Syncytial Virus G Glycoprotein-Induced Eosinophilia After Respiratory Syncytial Virus Challenge J. Immunol., February 15, 2003; 170(4): 2037 - 2045. [Abstract] [Full Text] [PDF]
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 F. P. Polack, M. N. Teng, P. L.Collins, G. A. Prince, M. Exner, H. Regele, D. D. Lirman, R. Rabold, S. J. Hoffman, C. L. Karp, et al. A Role for Immune Complexes in Enhanced Respiratory Syncytial Virus Disease J. Exp. Med., September 16, 2002; 196(6): 859 - 865. [Abstract] [Full Text] [PDF]
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 Y. Zhang, Y. Wang, X. Gilmore, K. Xu, P. R. Wyde, and I. N. Mbawuike An Aged Mouse Model for RSV Infection and Diminished CD8+ CTL Responses Experimental Biology and Medicine, February 1, 2002; 227(2): 133 - 140. [Abstract] [Full Text] [PDF]
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 G. A. Prince, S. J. Curtis, K. C. Yim, and D. D. Porter Vaccine-enhanced respiratory syncytial virus disease in cotton rats following immunization with Lot 100 or a newly prepared reference vaccine J. Gen. Virol., December 1, 2001; 82(12): 2881 - 2888. [Abstract] [Full Text] [PDF]
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L. M. Haynes, D. D. Moore, E. A. Kurt-Jones, R. W. Finberg, L. J. Anderson, and R. A. Tripp Involvement of Toll-Like Receptor 4 in Innate Immunity to Respiratory Syncytial Virus J. Virol., November 15, 2001; 75(22): 10730 - 10737. [Abstract] [Full Text] [PDF]
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T. R. Johnson, J. E. Fischer, and B. S. Graham Construction and characterization of recombinant vaccinia viruses co-expressing a respiratory syncytial virus protein and a cytokine J. Gen. Virol., September 1, 2001; 82(9): 2107 - 2116. [Abstract] [Full Text] [PDF]
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K. K. Tekkanat, H. F. Maassab, D. S. Cho, J. J. Lai, A. John, A. Berlin, M. H. Kaplan, and N. W. Lukacs IL-13-Induced Airway Hyperreactivity During Respiratory Syncytial Virus Infection Is STAT6 Dependent J. Immunol., March 1, 2001; 166(5): 3542 - 3548. [Abstract] [Full Text] [PDF]
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T. Hussell and P. J. M. Openshaw IL-12-Activated NK Cells Reduce Lung Eosinophilia to the Attachment Protein of Respiratory Syncytial Virus But Do Not Enhance the Severity of Illness in CD8 T Cell-Immunodeficient Conditions J. Immunol., December 15, 2000; 165(12): 7109 - 7115. [Abstract] [Full Text] [PDF]
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 P. J. M. OPENSHAW and C. HEWITT Protective and Harmful Effects of Viral Infections in Childhood on Wheezing Disorders and Asthma Am. J. Respir. Crit. Care Med., August 1, 2000; 162(2): S40 - 43. [Full Text] [PDF]
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 L. D. Hazlett, S. McClellan, B. Kwon, and R. Barrett Increased Severity of Pseudomonas aeruginosa Corneal Infection in Strains of Mice Designated as Th1 versus Th2 Responsive Invest. Ophthalmol. Vis. Sci., March 1, 2000; 41(3): 805 - 810. [Abstract] [Full Text]
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T. R. Johnson and B. S. Graham Secreted Respiratory Syncytial Virus G Glycoprotein Induces Interleukin-5 (IL-5), IL-13, and Eosinophilia by an IL-4-Independent Mechanism J. Virol., October 1, 1999; 73(10): 8485 - 8495. [Abstract] [Full Text] [PDF]
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A. Le Moine, M. Surquin, F. X. Demoor, J. C. Noel, M.-A. Nahori, M. Pretolani, V. Flamand, M. Y. Braun, M. Goldman, and D. Abramowicz IL-5 Mediates Eosinophilic Rejection of MHC Class II-Disparate Skin Allografts in Mice J. Immunol., October 1, 1999; 163(7): 3778 - 3784. [Abstract] [Full Text] [PDF]
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A. Srikiatkhachorn, W. Chang, and T. J. Braciale Induction of Th-1 and Th-2 Responses by Respiratory Syncytial Virus Attachment Glycoprotein Is Epitope and Major Histocompatibility Complex Independent J. Virol., August 1, 1999; 73(8): 6590 - 6597. [Abstract] [Full Text]
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