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* Division of Immunology, University of Connecticut Health Center, Farmington, CT 06030; and
Edward Jenner Institute for Vaccine Research, Compton, Newbury, Berkshire, United Kingdom
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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-, IL-4-, or IL-5-producing primary effector or memory CD4 T
cells was significantly higher in lung, liver, and intestinal lamina
propria (LP) as compared with spleen and lymph node. However, maximum
numbers of IL-4- and IL-5-producing cells were detected in the LP
several days after the peak of the Th1 response, and IL-5 production
was skewed toward the mucosal tissues. Remarkably, the recall response
resulted in sustained Th1 and Th2 responses in tertiary, but not
lymphoid tissues and long-term retention of Th1 and Th2 memory cells in
equal proportions in the LP. Finally, CD40 ligand was essential for
induction of IFN- in the spleen and LP, but not in the liver and
lung, while the IL-4 response required CD40 ligand only in the spleen.
Therefore, the rules governing the effector phenotype, and the overall
magnitude of the CD4 response, are regulated at the level of individual
tissues. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, and
lymphotoxin-
, which have been shown in some systems to be essential
in maintaining CD8 T cell effector functions (3). Th2
cells in contrast secrete cytokines such as IL-4, IL-5, and IL-10,
which induce class switching to IgE and IgG1 (1, 4). There
are likely to be other Th subsets, including Th0 cells, which are
considered to be Th1 and Th2 precursors, and produce both Th1- and
Th2-type cytokines (5). The balance between these types of
Th cells can influence the nature, strength, and duration of systemic
immune responses (6). Although the mechanisms by which CD4 T cells provide help to naive CD8 T cells are not fully understood, several studies have demonstrated that CD40-CD40 ligand (CD40L)3 interactions between dendritic cells (DC) and CD4 T cells activate the DC for effective priming of CD8 T cells (7, 8, 9). Interference with this pathway inhibits formation of germinal centers, Ab isotype switching, production of proinflammatory cytokines, NO, and IL-12, and CTL generation (10, 11). Thus, mice deficient in CD40 or CD40L are impaired in humoral and cell-mediated responses, although this is not absolute (12, 13, 14). Current models suggest that CD40L is induced mainly on CD4 T cells shortly after activation, and that engagement of CD40 expressed by APCs induces up-regulation of costimulatory molecules and production of proinflammatory cytokines (10). CD40 stimulation of DC induces IL-12 production, explaining the ability of this cell type to promote Th1 development (15, 16, 17), although other studies suggest CD40 may also regulate Th2 development (18). CD40L expressed by CD8 T cells also participates in immune responses. For example, optimal activation of CD8 T cells in the intestinal lamina propria (LP) requires CD40L expressed by CD8 T cells (19). Furthermore, the primary CD8 response to Listeria infection also requires CD40L/CD40 interactions, especially in the intestinal mucosa (20). Thus, costimulation and, potentially, CD4 T cell help can be mediated at the tissue level as well as the cellular level.
Recent studies have analyzed in detail the Ag-specific CD4 T cell
response to virus infection. In general, the CD4 response to virus
infection is numerically 
10- to 100-fold less than that of the CD8
response. Nevertheless, in the case of lymphocytic choriomeningitis
virus (LCMV) infection, long-lived splenic CD4 T cell memory is
induced, and although the primary and memory splenic responses are
quantitatively skewed toward CD4 T cells producing IFN-,
IL-4-producing cells are also detected throughout the response
(21). Although most studies have examined responses in
secondary lymphoid tissues, it has become increasingly clear that CD8
and CD4 T cell responses in nonlymphoid tissues constitute a large
portion of the overall response (22, 23, 24). It should also
be noted that adoptive transfer studies using in vitro activated CD4 T
cells suggest that Th1, but not Th2 cells are able to migrate to
tertiary tissues (25, 26). However, infection with
Nippostrongylus induces a strong Th2 response in the lung as
well as lymphoid tissues (27). In comparison with CD4
responses to viral and parasitic infections (28),
relatively little is known with regard to Ag-specific antibacterial CD4
T cell responses. One well-characterized model system used for
analyzing cellular immunity is infection with Listeria
monocytogenes (LM). Infection of mice with this pathogen induces a
strong T cell response that is essential for bacterial clearance from
the host. The hallmark of LM infection is a cell-mediated immune
response to secreted virulence factors inducing both CD8 and CD4 T cell
activation (29, 30, 31, 32, 33, 34, 35). Studies with MHC-deficient mice
reveal that both CD4 and CD8 T cells can contribute to protection
against listeriosis (36). Although LM has been suggested
to induce CD4 T cells that are restricted to the Th1 phenotype
(37, 38) and has been proposed as a vaccine vector for
immunity induction against HIV and tumors (39, 40),
relatively little is known about the generation and function of
LM-reactive CD4 T cells in vivo.
In this study, we report the characterization of the Ag-specific CD4 T cell response in secondary lymphoid and tertiary tissues following oral Listeria infection. The results showed that the nonlymphoid memory response outweighed that of the lymphoid response, and included Th1- and Th2-type cells. Furthermore, there were differential activation requirements for Th1 and Th2 CD4 T cells in different tissues.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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C57BL/6J and C57BL/6-CD40L-/- mice (41) were purchased from The Jackson Laboratory (Bar Harbor, ME).
Infection with rLM-OVA
A strain of rLM-secreting OVA (rLM-OVA) was produced, as
previously described, using a truncated OVA cDNA (20, 39, 42). Mice were infected by gavage with either 1 x
109 CFU rLM-OVA for primary infections or
1 x 106 CFU for initial priming,
followed by infection with 1–5 x 109
CFU to induce a recall response. Actual CFU used for infections were
calculated by plating dilutions of the inoculum. Bacterial titers
within tissues were determined by homogenizing the tissue in PBS
containing 1% saponin, plating serial dilutions of homogenate on
brain-heart infusion agar plates containing 5 µg/ml erythromycin, and
growing for 2 days at 37°C.
Isolation of lymphocyte populations
Single cell suspensions were prepared from lymph nodes and spleens. Lymphocytes were isolated from small intestine LP and liver, as previously described (23, 43). To obtain lymphocytes from lungs, anesthetized mice were perfused with PBS containing 75 U/ml heparin until the lungs were cleared of blood and white in color. The lungs were removed and cut into small pieces and stirred at 37°C for 30 min in HBSS containing 1.3 mM EDTA. Cells were washed in PBS containing 5% FCS twice. Lymphocytes were released from the tissue by digestion with 150 U/ml collagenase (Life Technologies, Rockville, MD) in RPMI containing 1 mM MgCl2, 1 mM CaCl2, and 5% FCS, at 37°C for 30 min. Released cells were pooled and then mashed through a cell strainer (BD Biosciences, Franklin Lakes, NJ). Cells were washed and resuspended in RPMI 1640 medium supplemented with 10% FCS at 1 x 107/ml.
Immunofluorescence analysis
At the indicated times after infection, lymphocytes were isolated, and OVA-specific CD8 T cells were detected using an H-2Kb tetramer containing the OVA-derived peptide SIINFEKL, produced as previously described (44, 45). For staining, lymphocytes were suspended in PBS/0.2% BSA/0.1% NaN3 (PBS/BSA/NaN3) at a concentration of 1 x 106–107 cells/ml, followed by incubation at room temperature for 1 h with OVA-tetramer-allophycocyanin plus the appropriate dilution of anti-CD8-PE (clone 53.6.7; BD PharMingen, San Diego, CA). Cells were washed with PBS/BSA/NaN3, stained with FITC-conjugated anti-CD11a and PerCP-conjugated anti-CD4 (clone RM4-5; BD PharMingen), incubated at 4°C for 20 min, washed, and fixed in 3% paraformaldehyde in PBS. Relative fluorescence intensities were measured with a FACSCalibur (BD Biosciences). Data were analyzed using WinMDI software (J. Trotter, The Scripps Clinic, La Jolla, CA).
ELISPOT assay
Cells secreting IFN-, IL-4, and IL-5 in an Ag-specific manner
were detected using a standard ELISPOT assay (46, 47).
Plates (Multiscreen HA plates; Millipore, Bedford, MA) were coated
overnight at 4°C with the cytokine-specific capture Abs specified
below, washed with PBS, then incubated with 200 µl RPMI 1640 medium
supplemented with 10% FCS for at least 1 h at room temperature.
Two-fold dilutions of cells from animals previously inoculated orally
with rLM-OVA were added to wells starting at
106–2 x 105
cells/well in the presence of 4 x 105
gamma-irradiated (1200 rad) syngeneic spleen cells. Cells were
incubated for 26 h with or without the OVA-derived peptide
TEWTSSNVMEERKIKV (OVA265–280) (48)
or the listeriolysin O (LLO)-derived peptide NEKYAQAYPNVS
(LLO190–201) (49). Peptides were
obtained from Research Genetics (Huntsville, AL). Wells were
sequentially washed three times each with ddH20, PBS, and PBS
containing 0.05% Tween 20, and then incubated for 20 h at 4°C
with biotinylated anti-cytokine Abs. Wells were washed and
incubated with peroxidase-labeled anti-biotin Ab (2 µg/ml; Vector
Laboratories, Burlingame, CA) for 20 h at 4°C. Wells were then
washed and spots were developed using freshly prepared substrate: 0.3
mg/ml 3-amino-9-ethyl-carbazole (Sigma-Aldrich, St. Louis, MO), 10%
dimethylformamide, 0.015%
H2O2 in 0.1 M sodium
acetate (pH 4.8). After 30 min, the substrate solution was discarded,
and plates were washed under running water and air dried. The following
combinations of capture and detection mAbs were used for IFN-, IL-4,
and IL-5, respectively: R46A2 (10 µg/ml) and XMG1.2-biotin (4
µg/ml), TRFK5 (4 µg/ml) and TRFK4-biotin (2 µg/ml), and BVD4-1D11
(2 µg/ml) and BVD4-24G2-biotin (2 µg/ml) (all Abs were from BD
PharMingen). Image analysis of colored spots was performed on an
Immunospot Image Analyzer (Cellular Technology, Cleveland, OH)
specifically designed for ELISPOT analysis. In all experiments, the
background spots (detected using effector cells plus feeders without
peptide) were subtracted from wells where spots were generated with the
addition of peptide. Statistics were performed using Student’s
t test.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We undertook a comparison of the CD8 and CD4 T cell responses in
tertiary vs lymphoid tissues after oral infection with rLM-OVA.
Consistent with our previous findings (20), the CD8
response, as detected using an MHC class I tetramer, peaked at day 9
postinfection, and a substantial population of OVA-specific CD8 T cells
was found in all tissues. The liver, lung, and intestinal LP contained
a larger population of tetramer+ cells, and the
response in these tissues was prolonged as compared with that of
secondary lymphoid organs (Fig. 1
A). To determine whether the
CD4 response followed similar kinetics, Ag-specific CD4 T cells were
quantitated from the same tissues using ELISPOT after stimulation with
an OVA-derived peptide presented by I-Ab
(48). Nine days after oral infection with rLM-OVA (the
peak of the response), all tissues contained substantial populations of
IFN-- and IL-4-producing OVA-specific cells (Fig. 1, B
and C). The liver, lung, and LP contained larger populations
of IFN--producing CD4 T cells (0.3, 0.4, and 0.4%, respectively)
compared with the spleen (0.1%) or mesenteric lymph node (MLN; 0.05%)
at the peak of the response (Fig. 1B). Interestingly, at the
same time point, the frequency of CD4 T cells producing IL-4 in the LP
(0.13%) was greater than in the spleen (0.08%), liver (0.09%), and
lung (0.08%). Even more intriguing was the finding that as the IFN-
response declined, the IL-4 response was sustained or increased in the
LP up to 15 days postinfection and did not fall to memory levels until
day 35.
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). When total numbers
of CD4 T cells producing IFN- were calculated, the cumulative sum of
the cell numbers in the LP, lung, and liver was similar to that in the
spleen at the peak of the response (Fig. 2). However, the total number
of CD4 T cells producing IL-4 in the spleen was greater than that in
the tertiary tissues 9 days postinfection. By day 15, a far greater
number of IL-4 producers was present in the tertiary tissues as
compared with the spleen, where the numbers had declined to memory
levels. This delayed IL-4 increase was attributed to the appearance of
significant numbers of IL-4-producing cells in the intestinal LP at day
15. Whether this reflects increased migration of cells to the LP or
continued expansion in the LP is unknown. Irrespective of this
increase, by 3–5 wk after infection, the total numbers of memory cells
producing each cytokine were similar between the tissues and the
spleen.
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-producing cells. Nine days after infection, greater
percentages of IFN--producing CD4 T cells were again detected in the
nonlymphoid tissues (Fig. 3). By day 15
postinfection, the IFN- response had declined in all tissues,
although less so in the LP. In the case of IL-5, the largest percentage
of IL-5-producing cells was present in the LP at day 9, but, as with
IL-4, this value had increased significantly by 15 days after
infection, and this was borne out when total cell numbers were
evaluated (data not shown). Unlike IL-4 producers, IL-5 producers had
also increased in the lung from 9 to 15 days postinfection. In the MLN,
spleen, and liver, fewer IL-5-producing cells were detected on day 9,
and these numbers had declined by day 15. These data suggested that
Listeria-specific Th2 CD4 T cells either preferentially
migrated to mucosal sites after primary activation or that
differentiation and expansion of Th2 cells occurred in those sites.
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To determine the localization pattern of CD4 memory cells
generated in response to Listeria infection,
cytokine-producing OVA-specific CD4 T cells were quantitated 42 days
after inoculation (Fig. 4).
Interestingly, the frequency of memory cells present in the tertiary
tissues as a whole was 10-fold greater than that found in the spleen
and MLN, implying a preferential retention and/or migration of memory
cells in nonlymphoid tissues (Fig. 4, A and B).
In the LP and the lung, roughly equal frequencies of IL-4-, IFN--,
and IL-5-producing CD4 T cells were detected. Fewer IFN-- and
IL-4-producing cells were found in the liver, while, as in the primary
response, IL-5 producers were barely detectable (Fig. 4A).
Analysis of total numbers of cytokine-producing memory cells indicated
that the MLN contained very few memory cells, while the spleen
contained numbers of memory cells comparable with those of individual
nonlymphoid tissues (Fig. 4C). As was observed in the
primary response, very few IL-5 producers were detected in the liver
(Fig. 4C). Overall, the cumulative sum of Ag-specific memory
cells in the tertiary tissues was greater than that present in the
secondary lymphoid tissues (Fig. 4D).
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). The overall response to the LLO
epitope was substantially larger than the OVA-directed response (Fig. 5A, day 9). Nevertheless, the general pattern of
cytokine-producing cells generated was qualitatively similar to that
observed in the OVA-specific response. Thus, at day 9 after infection,
a greater percentage of IFN--producing cells was detected in the LP
and the lung (although not liver) than in the spleen and MLN. Th2 cells
were again much more prominent in the mucosal tissues at the peak of
the response, and this was particularly true for IL-5-producing cells
in the lung and LP. Analysis of memory CD4 T cells 108 days
postinfection also reiterated the finding that substantially greater
frequencies of Ag-specific CD4 T cells producing either of the three
cytokines resided in the tertiary tissues compared with secondary
lymphoid tissues. Interestingly, in contrast to the IFN- response,
which had declined 10-fold in all tissues by this time, the Th2
response, as a percentage of CD4 T cells, had declined only 2- to
3-fold in the nonlymphoid tissues. These results indicated that, for
the most part, the character of the anti-OVA and anti-LLO CD4
responses was qualitatively similar.
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To examine the stability of the Listeria-specific CD4 T
cell response following a secondary infection, mice were infected
orally with 106 rLM-OVA, and at least 125 days
later were orally challenged with 5 x 109
bacteria and the OVA-specific response was measured. The small inoculum
used in the primary infection was necessary because oral infection with
high doses of Listeria resulted in significant protection
against oral reinfection, making it difficult to induce a detectable T
cell recall response (data not shown). By day 6 after reinfection, a
substantial number of CD4 T cells producing IFN- and IL-4 was
present in all tissues (Fig. 6),
indicating that a secondary, not a primary, response was being detected
because few Ag-specific CD4 T cells can be detected at this time point
during a primary infection. The Th1 response in all tissues peaked at
day 6, with the greatest frequency of IFN--producing CD4 T cells
present in the LP (0.6%). While the response in the MLN, spleen, and
lung declined to memory levels by day 20, the response in the liver and
LP was prolonged at least an additional 2 wk (Fig. 6A). In
fact, by day 60, the percentage of IFN- producers in the LP was
essentially unchanged from day 20, indicating the potential utility of
oral boosting for increasing intestinal protection. The peak of
IL-4-producing CD4 T cells after secondary infection was day 6 in the
MLN, spleen, and lung (0.06, 0.16, and 0.22%, respectively), while in
the LP and liver the response peaked at day 20 postinfection (0.32 and
0.35%, respectively) (Fig. 6B). At 60 days postinfection,
the frequency of IFN-- and IL-4-producing CD4 T cells in the lung
and liver was <0.05%, and in the MLN and spleen was <0.02% of CD4 T
cells. As was observed with the recall IFN- response, the frequency
of IL-4-producing CD4 T cells in the LP remained unchanged at 0.3%
of CD4 T cells from day 20 onward, suggesting that the LP is an
important reservoir for LM-specific CD4 memory T cells.
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CD40/CD40L interactions are critical for activation of CD4 T cells
and play a role in CD8 T cell activation in the intestinal LP
(19). Because the anti-Listeria CD4 T cell
response was differentially skewed in various tissues, we tested
whether CD40L was involved in this regulation by analyzing the response
in infected CD40L-deficient mice (Fig. 7). The Listeria-specific
primary IFN--producing CD4 T cell response in the spleen was
severely reduced in CD40L-/- mice compared with
the response in CD40L+/+ mice (Fig. 7, A and C), while the splenic Th2 response was
inhibited, but not as dramatically (Fig. 7, B and
D). However, although the LP Th1 response was
inhibited >80% in CD40L-/- mice, the LP Th2
response was unaffected. In striking contrast to the LP IFN-
response, CD40L was not required for the generation of Th1 CD4 T cells
in the lung or liver, nor was CD40L needed for the induction of Th2 CD4
T cells in these tissues (Fig. 7). Indeed, the Th1 and Th2 responses in
the lung were significantly increased in the absence of CD40L, perhaps
indicating a compensatory mechanism for decreased responses elsewhere
or a redistribution of activated cells.
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). Four days after infection, similar
numbers of bacteria were present in all tissues of control and
CD40L-/- mice. In the lungs of
CD40L-/- mice, 50% fewer bacteria were
found as compared with controls, perhaps as a result of the heightened
response in that tissue. At 9 days after infection, viable bacteria
were either not detected or were present in very small numbers in
either control or CD40L-/- mice. These results
demonstrated that CD40L was not required for bacterial clearance and
that IFN- responses in spleen and LP were also not critical for
resolution of infection in those tissues.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and 2). Furthermore, in the
primary as well as the memory response there was a much greater
frequency of IFN--, IL-4-, and IL-5-producing CD4 T cells in the LP,
lung, and liver as compared with the MLN and spleen. There are several
potential nonmutually exclusive reasons for the preferential
localization of effector and memory CD4 T cells in nonlymphoid tissues
as compared with secondary lymphoid tissues. The difference may reflect
enhanced survival in the nonlymphoid tissues, preferential homing to
those sites, selective retention of Ag-specific CD4 T cells, or greater
proliferation in the tertiary tissues, particularly in the primary
response. Homann et al. (21) recently reported that, while
LCMV-specific CD8 T cell memory was stably maintained for life, the
numbers of Ag-specific CD4 memory T cells gradually declined in the
spleen. However, other reports do not fully support this finding
(50, 51). Although we have not yet examined greatly
extended time periods, our findings suggested that while CD4 memory T
cells may decline in the lymphoid tissues, significant numbers of
memory cells remain in tertiary tissues. Such a scenario would imply
that the lymphoid pool of memory cells is regulated separately from the
tertiary tissue memory pool. Further in-depth migration studies will be
needed to address this important question. The substantial population
of responding extralymphoid CD4, as well as CD8, T cells should be
taken into account when analyzing immune responses.
Our data also demonstrated the capacity to induce both Th1- and
Th2-type Listeria-specific effector and memory cells, which
was heretofore unappreciated during LM infection. The conclusion that
IL-4 was not produced by conventional T cells in LM infection is
derived from studies using i.v. or i.p. routes of infection, followed
by restimulation of unseparated spleen cells in vitro and measurement
of cytokines by ELISA (38, 52), or from PCR analysis of
whole tissue lysates (53). Restimulation was achieved
using either heat-killed LM (52) or LM culture
supernatants (38). IFN-, but not IL-4, was readily
detected in these systems. It is not clear in these experiments whether
sufficient quantities of the relevant antigenic epitopes are available
after pulsing APC in vitro with whole killed organisms or culture
supernatants. The timing or sensitivity of the assays used or perhaps
the use of IL-4 in the cultures may also influence the results.
Nonetheless, as we demonstrated in this study, similar numbers of IL-4-
and IFN--producing cells are found in the spleen at the peak of the
primary response. The relevant quantity of a given cytokine in an
immune response in vivo is difficult to determine, but, in any case,
the ELISPOT assay using defined MHC class II-presented peptides for
restimulation allowed us to quantitate the number of IL-4- and
IL-5-producing cells in a number of tissues. The IL-4 producers were
conventional MHC class II-restricted CD4 T cells because when CD4 T
cells were depleted from lymphocytes before plating or in the absence
of peptide or after infection of MHC class
II-/- mice, few IL-4-producing cells were
detected at the time points tested (data not shown). In contrast, the
very early response to LM infection is characterized by rapid
production of IL-4 by CD1-restricted NK-T cells in the liver (52, 54). Therefore, it is possible that early IL-4 production plays
a role in driving a subset of Ag-specific CD4 T cells toward the Th2
phenotype during the priming phase (55). However, recent
work also suggests that early production of IL-4 in an immune response
in vivo may direct the response down a Th1 pathway (56).
Thus, given the complexity of the innate and adaptive immune components
involved in LM infection, it is perhaps not surprising that the CD4 T
cell response does not strictly adhere to the conventional Th1/Th2
paradigm. Similarly, following acute infection with LCMV, a mixed CD4 T
cell primary and memory response is induced in the lymphoid tissues
(50).
While it is clear that IFN- plays an important role in protection
against LM infection (56), the role of IL-4 or IL-5
derived from conventional T cells has not been determined. Blocking of
IL-4 during LM infection results in increased resistance
(57), but the source and timing of production of the
relevant IL-4 being inhibited are unknown (e.g., IL-4 production by
NK/NK-T cells, conventional CD4 T cells, or nonhemopoietic cells). The
substantial numbers of IL-4- and IL-5-producing effector and memory
cells in the LP and lung could be involved in regulation of Ab
production in those tissues or in LN draining those sites. The
localization of Th1- and Th2-type memory cells in the nonlymphoid
tissues may also be important for a rapid recall response in situ.
Indeed, oral reinfection resulted in a sustained increase in memory
cells in the LP, but not in the spleen or LN (Fig. 6). The LM-specific
primary and recall CD8 response is also sustained in the LP
(20). Thus, CD4 T cells in the LM response may also be
important for magnification and modulation of the CD8 T cell response
in nonlymphoid tissues. This hypothesis is supported by previous
studies showing that CD4 and CD8 T cells cooperate in mounting a
protective anti-LM immune response (58).
Tissue-level control of Th1/Th2 skewing was exemplified by the
requirement for CD40L in differentially regulating the response in
different tissues. Remarkably, while the overall splenic CD4 T cell
response was inhibited in the absence of CD40L, the induction of
IFN- producers, and not IL-4 producers, was blocked in the
intestinal LP without CD40L. Moreover, both the Th1 and Th2 responses
in the lung were in fact augmented in CD40L-/-
mice, while the liver and MLN responses were unaffected. In comparison,
we previously showed that the splenic anti-LM CD8 T cell response
was largely unaffected, while the LP CD8 response was inhibited in the
absence of CD40/CD40L interactions (20). Overall, these
results demonstrated a conspicuous tissue and cell type-specific
regulation and implied that costimulation via CD40L was differentially
used depending on the tissue. The influence of CD40/CD40L interactions
on helper differentiation is somewhat controversial, although it has
been suggested that the CD40 pathway is more important for Th1
development (59). Activation in vitro of
CD40L-/- TCR transgenic CD4 T cells results in
decreased IFN- production, but, interestingly, an increase in IL-4
production, in accordance with what we observed in the lung
(60). Consistent with our findings in the spleen, both Th1
and Th2 LCMV-specific splenic responses in
CD40L-/- mice were reduced (61).
CD40/CD40L interactions are also critical in protection against
Leishmania major infection via up-regulation of IL-12 and
IFN- production (12, 13, 14). In contrast, CD40/CD40L
interactions are not essential for Th1-mediated protection against
infection with the intracellular pathogens Histoplasma
(62) or Mycobacterium (63).
Similarly, while IFN- is involved in protection against LM
infection, the inhibition of IFN- in spleen and LP, which we
observed in CD40L-/- mice, did not affect
clearance of the bacteria. It is interesting to speculate that
sufficient levels of IFN- were produced in other tissues to afford
protection or that other mechanisms can come into play in the absence
of IFN-.
In summary, the data presented bolster the hypothesis that T cell responses can be compartmentalized at the tissue level, even during a systemic infection. By the same token, the distinct response type in a given tissue is likely to be important in mediating or regulating immunity at that site.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Leo Lefrançois, Department of Medicine, University of Connecticut Health Center, M/C 1319, 263 Farmington Avenue, Farmington, CT 06030. E-mail address: llefranc{at}neuron.uchc.edu
3 Abbreviations used in this paper: CD40L, CD40 ligand; DC, dendritic cell; LCMV, lymphocytic choriomeningitis virus; LLO, listeriolysin O; LM, Listeria monocytogenes; LP, lamina propria; MLN, mesenteric lymph node.
Received for publication January 9, 2002. Accepted for publication March 4, 2002.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-galactosylceramide directs conventional T cells to the acquisition of a Th2 phenotype. J. Immunol. 163:2373.This article has been cited by other articles:
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 M. Deckert, S. Virna, M. Sakowicz-Burkiewicz, S. Lutjen, S. Soltek, H. Bluethmann, and D. Schluter Interleukin-1 Receptor Type 1 Is Essential for Control of Cerebral but Not Systemic Listeriosis Am. J. Pathol., March 1, 2007; 170(3): 990 - 1002. [Abstract] [Full Text] [PDF]
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J. P. Hewitson, G. R. Jenkins, P. A. Hamblin, and A. P. Mountford CD40/CD154 Interactions Are Required for the Optimal Maturation of Skin-Derived APCs and the Induction of Helminth-Specific IFN-{gamma} but Not IL-4. J. Immunol., September 1, 2006; 177(5): 3209 - 3217. [Abstract] [Full Text] [PDF]
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C. Zaph, K. A. Rook, M. Goldschmidt, M. Mohrs, P. Scott, and D. Artis Persistence and Function of Central and Effector Memory CD4+ T Cells following Infection with a Gastrointestinal Helminth J. Immunol., July 1, 2006; 177(1): 511 - 518. [Abstract] [Full Text] [PDF]
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D. M. Lindell, T. A. Moore, R. A. McDonald, G. B. Toews, and G. B. Huffnagle Distinct Compartmentalization of CD4+ T-Cell Effector Function Versus Proliferative Capacity during Pulmonary Cryptococcosis Am. J. Pathol., March 1, 2006; 168(3): 847 - 855. [Abstract] [Full Text] [PDF]
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J. K. Whitmire, N. Benning, and J. L. Whitton Precursor Frequency, Nonlinear Proliferation, and Functional Maturation of Virus-Specific CD4+ T Cells. J. Immunol., March 1, 2006; 176(5): 3028 - 3036. [Abstract] [Full Text] [PDF]
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K. Kalies, M. Blessenohl, J. Nietsch, and J. Westermann T Cell Zones of Lymphoid Organs Constitutively Express Th1 Cytokine mRNA: Specific Changes during the Early Phase of an Immune Response J. Immunol., January 15, 2006; 176(2): 741 - 749. [Abstract] [Full Text] [PDF]
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M. M. Freeman and H. K. Ziegler Simultaneous Th1-Type Cytokine Expression Is a Signature of Peritoneal CD4+ Lymphocytes Responding to Infection with Listeria monocytogenes J. Immunol., July 1, 2005; 175(1): 394 - 403. [Abstract] [Full Text] [PDF]
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 A. C. Kirby, M. Sundquist, and M. J. Wick In Vivo Compartmentalization of Functionally Distinct, Rapidly Responsive Antigen-Specific T-Cell Populations in DNA-Immunized or Salmonella enterica Serovar Typhimurium-Infected Mice Infect. Immun., November 1, 2004; 72(11): 6390 - 6400. [Abstract] [Full Text] [PDF]
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Z. Liu and L. Lefrancois Intestinal Epithelial Antigen Induces Mucosal CD8 T Cell Tolerance, Activation, and Inflammatory Response J. Immunol., October 1, 2004; 173(7): 4324 - 4330. [Abstract] [Full Text] [PDF]
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M. J. Montfort, H. G. A. Bouwer, C. R. Wagner, and D. J. Hinrichs The Development of Functional CD8 T Cell Memory after Listeria monocytogenes Infection Is Not Dependent on CD40 J. Immunol., September 15, 2004; 173(6): 4084 - 4090. [Abstract] [Full Text] [PDF]
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A. L. Marzo, V. Vezys, K. D. Klonowski, S.-J. Lee, G. Muralimohan, M. Moore, D. F. Tough, and L. Lefrancois Fully Functional Memory CD8 T Cells in the Absence of CD4 T Cells J. Immunol., July 15, 2004; 173(2): 969 - 975. [Abstract] [Full Text] [PDF]
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M. Morimoto, M. Morimoto, J. Whitmire, S. Xiao, R. M. Anthony, H. Mirakami, R. A. Star, J. F. Urban Jr, and W. C. Gause Peripheral CD4 T Cells Rapidly Accumulate at the Host:Parasite Interface during an Inflammatory Th2 Memory Response J. Immunol., February 15, 2004; 172(4): 2424 - 2430. [Abstract] [Full Text] [PDF]
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B. Spellberg, D. Johnston, Q. T. Phan, J. E. Edwards Jr., S. W. French, A. S. Ibrahim, and S. G. Filler Parenchymal Organ, and Not Splenic, Immunity Correlates with Host Survival during Disseminated Candidiasis Infect. Immun., October 1, 2003; 71(10): 5756 - 5764. [Abstract] [Full Text] [PDF]
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L. Lefrancois, A. Marzo, and K. Williams Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo J. Immunol., September 15, 2003; 171(6): 2832 - 2839. [Abstract] [Full Text] [PDF]
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H. Saklani-Jusforgues, E. Fontan, N. Soussi, G. Milon, and P. L. Goossens Enteral Immunization with Attenuated Recombinant Listeria monocytogenes as a Live Vaccine Vector: Organ-Dependent Dynamics of CD4 T Lymphocytes Reactive to a Leishmania major Tracer Epitope Infect. Immun., March 1, 2003; 71(3): 1083 - 1090. [Abstract] [Full Text] [PDF]
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D. J. Shedlock, J. K. Whitmire, J. Tan, A. S. MacDonald, R. Ahmed, and H. Shen Role of CD4 T Cell Help and Costimulation in CD8 T Cell Responses During Listeria monocytogenes Infection J. Immunol., February 15, 2003; 170(4): 2053 - 2063. [Abstract] [Full Text] [PDF]
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F. Sisto, A. Miluzio, O. Leopardi, M. Mirra, J. R. Boelaert, and D. Taramelli Differential Cytokine Pattern in the Spleens and Livers of BALB/c Mice Infected with Penicillium marneffei: Protective Role of Gamma Interferon Infect. Immun., January 1, 2003; 71(1): 465 - 473. [Abstract] [Full Text] [PDF]
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