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Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada
| Abstract |
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| Introduction |
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14-kDa protein by transfected COS-7 and
BW5147 cells (6). Although they share only limited protein homology
(
30%), IL-4 and IL-13 belong to the same
-helix protein family
(5, 7). The IL-4R
-chain of the IL-4R is a component of some but not
all IL-13 receptors (8, 9). In addition, two IL-13-binding receptor
chains have been identified: IL-13R
1 (NR4; Refs. 8 and 1012) and
IL-13R
2 (13), which share high levels of sequence identity with
known cytokine receptors. Four structural models of IL-13R involving
different combinations of IL-13R
1, IL-13R
2, or IL-4R
have been
proposed (14). The sharing of a receptor component may explain why a
mutant IL-4 protein acts as a competitive receptor antagonist to both
IL-4 and IL-13 (5, 7). Consistent with their protein homology and
partial receptor sharing, the functions of IL-13 partially overlap with
those of IL-4. IL-13 exhibits pleiotropic effects on human peripheral blood monocytes and mouse granulocyte-macrophage CSF-derived bone marrow macrophages. IL-13 (in common with IL-4) induces morphological and surface Ag changes in human monocytes and mouse granulocyte-macrophage CSF-derived bone marrow macrophages, and also inhibits Ab-dependent cellular cytotoxicity activity and production of nitric oxide and proinflammatory cytokines (2, 3, 15, 16) without affecting the phagocytic functions of activated macrophages (16). In addition, IL-13 inhibits the replication of HIV in monocytes (17) and enhances VCAM-1 expression on human endothelial cells in vitro (18). Consistent with the ability of IL-13 to stimulate early hemopoietic precursors in vitro (19), IL-13 administration in vivo also induces extramedullary hemopoiesis and monocytosis in mice (6).
Human IL-13 and IL-4 have similar effects on human B cells, although IL-4 is generally more potent. IL-13 enhances proliferation of anti-IgM- or anti-CD40-activated human B cells (3, 20, 21), modulates their surface Ags (2, 3, 22), and induces class switching to IgG4 and IgE synthesis in human B cells in combination with activated T cells, anti-Ig, anti-CD40 Abs, or CD40 ligand (CD40L; 2022). However, IL-13 does not have any additive or synergistic effects with IL-4 on induction of IgG4 and IgE (20, 22). In contrast to hIL-13 effects on human B cells, mIL-13 has been reported to have no effects on mouse B cells (5). Although this is consistent with the observation that IL-4-deficient mice do not produce IgE in response to nematode infection (23), these mice produce IgE upon malaria infection (24). This finding suggests that an IL-4-independent mechanism for IgE synthesis also exists in mice and raises the possibility that mIL-13 may also induce IgE synthesis in some circumstances.
As IL-13 is associated with the Th2 response in mice, which includes strong Ab production, and hIL-13 stimulates Ab production by human B cells, we have re-examined the potential effects of mIL-13 on mouse Ab production. We found that IL-13 stimulated Ab production in vivo during a strong immune response. Analysis of mouse B cells in vitro also showed that IL-13 stimulated Ab production by B cells that were activated by various stimuli, including T cells or anti-CD40 Ab. IL-13 was less effective than IL-4 and did not show the strong selectivity for IgG1 and IgE that is characteristic of IL-4. Nevertheless, in the presence of low doses of anti-CD40, IL-13 directly stimulated sIgD+ B cells, resulting in increased numbers of Ab-producing B cells as measured by enzyme-linked immunospot (ELISPOT) assay. Similar findings were obtained in the presence of neutralizing IL-4 mAb, suggesting that the effect of IL-13 was independent of IL-4. This enhancement appears to be due to increased survival, but not proliferation, of mouse B cells. Thus, IL-13 can also contribute to B cell Ab production in mice, consistent with some of the known effects of IL-13 on human B cells.
| Materials and Methods |
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Recombinant mIL-13 was purified using Mono Q 16/10 anion
exchange column (Pharmacia Biotechnology, Uppsala, Sweden), as
previously described, from the supernatant of mouse BW5147 cells stably
transfected with mIL-13 (6). Fractions from the peak two were
then pooled, equilibrated in 0.1% trifluoroacetic
acid-H2O, and further purified using a RESOURCE
RPC reverse-phase column (Pharmacia Biotechnology) eluting with a
gradient of 070% acetronitrile-0.1% trifluoroacetic acid. Fractions
were screened for IL-13 bioactivity on TF-1 cells, and positive
fractions were equilibrated in PBS as described (6). The purity of
IL-13 was assessed by 4 M urea urea-15% SDS-PAGE. Both Coomassie blue
staining and Western blot analysis using anti-mouse-IL-13 (RAMP1)
Ab demonstrated that the purified IL-13 migrated as an
14 kDa
doublet, similar to the native IL-13 secreted by D10.G4.1, a Th2 cell
line. In addition to the doublet, Western blot analysis and Coomassie
staining also recognized small amounts of heterogenous material at
higher m.w., which may be due to carbohydrate heterogeneity on IL-13.
Subsequent experiments were performed with this purified IL-13 with sp.
act. of 3.6 U/ng (6). Purified IL-13 contained <0.1 ng of endotoxin
per 400 µg of IL-13, as determined by the Limulus assay
(Sigma, St. Louis, MO).
In vivo IL-13 treatment
BALB/c female mice (68 wk old) were obtained from Health Sciences Laboratory Animal Services (HSLAS, Edmonton, Alberta, Canada). PBS controls or IL-13 were coded and loaded into Alzet microosmotic pumps (Alza, Palo Alto, CA), which were then implanted into the peritoneal cavity through a small dorsal incision as previously described (6). Three or four mice were implanted with pumps containing PBS or each dose of IL-13. Four hours later, each mouse was immunized i.p. with 0.2 ml of 50% chicken RBCs (CRBCs) resuspended in saline. The mice were maintained in the HSLAS animal facility according to the guidelines of the Canadian Council on Animal Care and monitored daily for any abnormality. On day 7, the mice were anesthetized, and blood was collected by cardiac puncture in uncoated or heparin-coated syringes. The mice were sacrificed, and their spleens were collected and weighed.
Hemagglutination assay
Direct and indirect hemagglutination assays were performed as described (25, 26). Briefly, wells of round-bottom 96-well plates containing 100 µl of doubling dilutions of plasma in PNS buffer (0.05 M phosphate, 0.1 M NaCl, 1% FBS) were incubated with 100 µl of 0.5% CRBCs for a minimum of 45 min, after which the wells were scored for direct hemagglutination titer. To obtain indirect hemagglutination titers, the cells were washed three times with PNS and 200 µl of a 1:1000 dilution of anti-mouse IgG1, IgG2a, IgG2b, IgG3 (Sigma), or Ig pan-specific (PharMingen, San Diego, CA) Abs were added. Titers were expressed as the reciprocal of the last dilution that gave positive CRBCs agglutination.
B cell purification
B220+ or sIgD+ B cells were obtained as previously described (27). Briefly, nonadherent spleen cells were obtained from 8- to 12-wk-old female BALB/c mice (HSLAS), stained with phycoerythrin-conjugated anti-CD4 and anti-CD8 and FITC-conjugated anti-B220 or anti-IgD (PharMingen) on ice for 30 min, and washed. After purification by Lympholyte-M (Cedarlane Laboratories, Hornby, Ontario, Canada), the lymphocytes were sorted for B220+ or sIgD+ cells using a Coulter EPICS Elite ESP cell sorter (Beckman Coulter, Hialeah, FL). The purity of the sorted cells was verified to be >99.5% by FACScan analysis (Becton Dickinson, San Jose, CA).
B cell stimulation by fixed T cells
T cell lines were maintained as described previously (28). After being coated overnight at 4°C or room temperature for 4 h with purified anti-CD3 (145-2C11, provided by J. Bluestone, University of Chicago, Chicago, IL) in PBS, polystyrene plates were rinsed with PBS, and T cells were added at a density of 1 x 106 cells/ml for 24-h activation. Anti-CD3-activated T cells were collected, washed three times with PBS, and fixed with 0.4% paraformaldehyde (PFA) for 5 min (29, 30). The fixed anti-CD3-activated T cells were washed with PBS and resuspended in RPMI 1640 (Life Technologies, Burlington, Ontario, Canada) before being added to cultures. Sorted B cells (3 to 5 x 104 cells/well) with at least 99% purity were cultured for 7 days with different ratios of 0.4% PFA-fixed anti-CD3-activated M264-15 or D10.G4.1 cells. Ab levels were measured by ELISA as described below.
B cell stimulation by anti-CD40 Ab
Sorted sIgD+ B cells (5 x 104 cells/well) with >98% purity were cultured with various concentrations of anti-CD40 Ab (PharMingen), with or without purified recombinant mIL-4 (a generous gift from Schering-Plough, Kenilworth, NJ), IL-13, or anti-IL-4 mAb (11B11). Iscoves modified Dulbeccos medium (IMDM, Life Technologies) containing 8% FCS (Hy-Clone, Logan, UT), 5 µg/ml of gentamicin sulfate (Life Technologies), and 50 µM of 2-ME (Sigma) was used in all anti-CD40 cultures. Supernatants were harvested after 67 days of culturing, and Ig levels were quantitated as described below. For microcultures, the bulk cultures were serially diluted, and 10 µl of the cultures with various concentrations of anti-CD40 Ab, with or without purified IL-4, IL-13, or anti-IL-4 mAb (11B11), containing 90100 sorted sIgD+ or B220+ B cells were cultured in Terasaki 96-well trays (Robbins Scientific, Sunnyvale, CA). The Terasaki trays were incubated in moist containers, and B cells in each well were enumerated after culturing for 2 days.
Determination of cell surface molecules
Sorted sIgD+ B cells (5 x 104 cells/ml) were cultured with 1 or 3 ng/ml of anti-CD40 Ab, with or without purified mouse IL-4 or IL-13, for 2 days. Cells were harvested, enumerated, and stained with Abs to MHC class II, MHC class I, CD23, and IgM (all from PharMingen). Cells (25,000/population) were analyzed by a FACScan using CellQuest software (Becton Dickinson).
ELISA assay for Igs
All Abs were used at 1:1000 dilution unless stated otherwise.
Affinity-purified goat anti-mouse IgG1, IgG2a, IgG2b, IgG3, IgA, or
IgM Abs (Sigma) in PBS were used to coat Falcon 3911 microtiter plates
(Becton Dickinson Labware, Oxnard, CA) at 4°C overnight. Each coating
Ab exhibited minimal cross-reactivity with other Ig subclasses. After
washing the plates, dilutions of plasma or culture supernatants were
added to the wells and incubated for at least 30 min at room
temperature. Peroxidase-conjugated polyvalent anti-mouse Ig was
used for detection (Sigma). Purified mouse IgG1, IgG2a, IgG2b, IgG3,
and IgM (Sigma) were used as standards. Mouse IgE levels were
quantitated in comparison to purified mouse IgE
by two-site sandwich
ELISA (PharMingen), as previously described (6) and according to the
manufacturers recommendations.
ELISPOT assay for mouse IgM production
B cells from each Terasaki well were transferred to MultiScreen 96-well plates (Millipore, Bedford, MA) that were coated with affinity-purified goat anti-mouse IgM Ab at 1:1000 dilution (Sigma). After overnight incubation, the plates were developed for Ab spots. After each step, the incubation buffers were filtered and washed with 0.05% Tween 20 (Sigma) in PBS. Each well was incubated with biotinylated affinity-purified goat anti-mouse Ig resuspended in PBS buffer containing 1% FCS and 1% Tween 20 for 1 h. After being incubated with peroxidase-conjugated strepavidin (Jackson ImmunoResearch Laboratories, West Grove, PA), the plates were washed and developed with AEC substrate (Vector Laboratories, Burlingame, CA) according to the manufacturers instructions. The plates were dried, and Ab spots were enumerated.
B cell proliferation assay
Sorted sIgD+ B cells (5 x 104 or 1 x 104 cells/well) were cultured with various concentrations of anti-CD40 Ab, with or without purified IL-13 or IL-4, for 3 days. A total of 1 µCi/well of [3H] (Amersham, Arlington Heights, IL) was added during the final 18 h of culture. Cells were harvested using a MicroMate 196 cell harvester (Canberra Packard, Meriden, CT), and radioactive incorporation was determined on a Matrix 96 (Canberra Packard).
Determination of B cell division
Sorted sIgD+ B cells were stained with 5-(and 6-)carboxyfluorescein diacetate, succinimidyl ester (CFSE, Molecular Probes, Eugene, OR) as described (31). Briefly, the cells were washed with IMDM and stained with 0.11 µM CFSE for 10 min. At the end of the incubation, the cells were washed three times with ice-cold IMDM containing 8% FCS. The stained cells (at a density of 1.25 x 105 c/ml) were cultured in a 24-well Falcon plate (Becton Dickinson Labware) in the presence of 3 ng/ml of anti-CD40, with or without 520 ng/ml of IL-13 or 10 ng/ml of IL-4. On day 3, the cells were harvested and washed, and viable cells were counted using Trypan Blue. Cell division was assessed by flow cytometry analysis of 25,000 cells/population using a FACScan and CellQuest software (Becton Dickinson). Dead cells were excluded based on their light scattering properties.
Statistical analyses
The unpaired Students two-tailed t test was used to calculate p values. Alternatively, the Kruskal-Wallis test was used to calculate p values for serum CRBCs agglutination titers.
| Results |
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To study the effects of IL-13 during a strong immune response, IL-13 or PBS was administered via osmotic pump, implanted into the peritoneal cavity of BALB/c female mice, over a period of 7 days. Dosages of IL-13 ranging from 0.5 to 6.5 µg/mouse/day were administered. After the pump implantation, the mice were immunized with CRBCs in the peritoneal cavity. Consistent with our previous observations (6), in vivo IL-13 administration induced splenomegaly (Expt. 2: 2-fold, p < 0.03; and Expt. 3: 1.5-fold, p < 0.04) due to increased splenocyte numbers, even during the strong immune response to CRBCs. PBS-treated mice immunized with CRBCs showed an approximate 2-fold increase of total plasma IgG1, IgG2a, IgG2b, IgG3, and IgM compared with untreated and nonimmunized littermates (data not shown).
When CRBC-immunized mice were treated continuously with different doses
of IL-13 in three separate experiments, significant further increases
in total Ig levels were observed for IgG1, IgG2a, and IgG2b (Fig. 1
). IgM and IgG3 levels showed a
significant increase in only one of the three experiments, and IgA did
not show any significant changes. In one experiment (Expt. 3), IgE
levels (IL-13, 137 ± 82.3 vs PBS, 176.6 ± 93.5) were not
significantly different between IL-13 and control groups. Thus, in vivo
administration of IL-13 enhanced total serum or plasma levels of at
least three IgG subclasses during a strong immune response to CRBCs.
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The indirect and direct hemagglutination titers in the plasma of
these mice were assayed to quantitate Ag-specific Ab production in
vivo. The IL-13-treated groups consistently showed increased direct and
indirect CRBCs agglutination titers compared with the PBS-treated
groups (Fig. 2
A).
Isotype-specific indirect CRBCs agglutination titers of IgG1, IgG2a,
and IgG2b were also elevated in the IL-13-treated group (Fig. 2
B). Hence, in vivo treatment with IL-13 enhanced the
Ag-specific Ab response.
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Collectively, these data indicate that in vivo IL-13 treatment induced higher levels of IgG1, IgG2a, and IgG2b in vivo. These in vivo effects could have been due to direct or indirect effects of IL-13. As IL-13 treatment in vivo did not alter the levels of cytokines secreted by Con A-stimulated spleen cells in vitro (data not shown), we hypothesized that mouse B cells might respond directly to mIL-13, analogous to the direct effects of hIL-13 on human B cells (2, 3, 20, 21, 22). To investigate the effects of IL-13 on mouse B cells, different numbers of PFA-fixed anti-CD3-activated D10.G4.1 (Th2) T cells were cultured with B220+ splenic B cells in the absence or presence of varying doses of IL-13 or IL-4 for 7 days. T cells fixed after activation provide a source of CD40L, which is expressed on activation and is the major cell-surface activating signal delivered by T cells to B cells (29, 30, 32, 33, 34, 35, 36, 37, 38, 39).
Both IL-13 and IL-4 consistently enhanced the levels of IgM and IgG1 in
the supernatant (Fig. 3
). However,
compared with IL-4, IL-13 was consistently at least 2-fold less
effective in enhancing IgM production and more than 10-fold less
effective on IgG1 synthesis. In most experiments, IL-13 also showed at
least additive effects in stimulating Ab production by B cells
stimulated with Th1 clones, irradiated primary alloreactive Th1 cells,
or CD40L-transfected cells (data not shown). IL-13 effects on the
production of IgG2a, IgG2b, and IgG3 varied between experiments,
possibly due to the release of endogenous cytokines such as IFN-
,
IL-4, or IL-13 from the fixed T cells.
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Although IL-13 enhanced Ig production in bulk culture, and the
sIgD+ sorted mouse B cells were >99.5% pure, each well
contained 50,000 cells. Thus, up to 250 contaminating cells could have
been present per well, and indirect effects of IL-13 were theoretically
possible. To exclude this possibility, sIgD+ sorted mouse B
cells were titrated to 90100 cells per well and cultured in various
combinations of anti-CD40 mAb, IL-13, and IL-4. Under these
conditions, at least some wells must have contained only B cells,
allowing a stringent test of the ability of IL-13 to enhance Ab
production in the absence of any other cell type. After 2 days, the
cells in each well were counted and transferred to affinity-purified
goat anti-mouse IgM-coated nitrocellulose plates for an ELISPOT
assay. In the presence of anti-CD40 mAb, different doses of IL-13
not only enhanced the number of B cells that survived after 2 days of
culture but also increased the number of IgM-secreting B cells (Fig. 6
). Similar data were obtained in eight
separate experiments (three are shown in Fig. 6
). Unlike IL-13-treated
cells, IL-4 and anti-CD40-responsive sIgD+ B cells
formed tight clusters, and the number of Ab spots could not be
accurately enumerated, supporting the idea that IL-4 and IL-13 have
different effects on mouse B cells.
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The increased numbers of B cells in the microwell experiments
indicated that IL-13 enhanced either the survival or proliferation of
mouse B cells. To evaluate the effects of IL-13 on proliferation,
sIgD+ splenic mouse B cells were stimulated with
anti-CD40 Ab in the absence or presence of IL-13 or IL-4, and
thymidine incorporation was measured on day 3. IL-4 strongly enhanced
proliferation of mouse B cells, particularly in the presence of
anti-CD40 Ab (Fig. 8
). However, IL-13
did not consistently enhance the proliferation of
anti-CD40-stimulated sIgD+ B cells, although slight
effects were seen in some experiments (e.g., Fig. 8
).
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| Discussion |
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In agreement with previous reports (5, 41), we also found that IL-13 had no detectable effect on LPS-stimulated mouse B cells, and B cells stimulated strongly by anti-CD40 Ab (>100 ng/ml) also showed no additional response to IL-13 (data not shown). Hence, the biological effects of IL-13 on mouse B cells may not be revealed during potent stimulation by either anti-CD40 Ab or LPS. We have now shown that IL-13 has biological effects on mouse B cells at suboptimal doses of anti-CD40 Ab, which may represent more physiological stimulation levels. Thus, effects of IL-13 may not be observed in all B cell stimulatory systems, which may account for a previous report that IL-13 failed to exhibit any biological functions on mouse B cells (5).
IL-4 enhances proliferation of mouse and human B cells and moderately stimulates synthesis of most Ig subclasses. In addition, IL-4 strongly stimulates synthesis of IgG1 (mouse) or IgG4 (human) and IgE (mouse and human) by the induction of specific class switching (42, 43, 44, 45). hIL-13 stimulates sIgD+ B cells to produce IgG4 and IgE (20, 21, 22). However, our results with mIL-13 suggested that although this cytokine may share some of the general B cell stimulatory effects of IL-4, there is no strong isotype selectivity. IgG1 levels were enhanced only to a similar extent as other isotypes, and we were unable to detect significant levels of IgE production when B cells were stimulated by IL-13 and fixed, activated T cells or anti-CD40 Ab, although IL-4 induced high levels of IgE in both conditions. If IgE production is stimulated by IL-13, the levels are below our detection limit (6.25 ng/ml), or at least 50-fold below the levels induced by IL-4. The difference between IL-4- and IL-13-induced levels is more than 10-fold for IgG1, thus it remains possible that IL-13 may indirectly enhance IgE levels to a low extent, by increasing B cell survival, for example.
Interestingly, IgE was produced in IL-4-deficient mice infected with malaria (24) or Leishmania major (46) or during the course of retrovirus-induced immunodeficiency syndrome or anti-IgD treatment (47), but not during nematode infection (23, 47, 48), suggesting that there is an IL-4-independent mechanism that can also stimulate IgE production in some circumstances. Although our results do not provide any evidence for a selective role of IL-13 in IgE production, we cannot yet exclude the possibility that IL-13 may enhance IgE levels under certain conditions that we have not yet tested.
The enhancement of IgE synthesis by IL-13 in human but not mouse B cells suggests a species difference in the requirements for IgE class switching. Although many cytokine effects are conserved between mice and humans, some differences have been reported. Mouse bone marrow cells cultured with mIL-3 for >4 wk produce almost pure mouse-mast cell populations (49), whereas hIL-3 induces production of basophils and eosinophils but not mast cells from human cord blood or bone marrow mononuclear cells (50). Moreover, mIL-5 is a potent B cell stimulator, but hIL-5 does not appear to be a regulatory factor for human B cells except in the presence of mitogenic stimuli such as Moraxella catarrhalis (51) or Staphylococcus aureus Cowan I strain (SAC) under particular conditions (52). Also, IL-10 stimulates proliferation of SAC-stimulated human B cells (53), but inhibits proliferation of LPS-stimulated mouse B cells (54).
The capacity of IL-4 and IL-13 to enhance DNA synthesis and
proliferation of human B cells stimulated with anti-IgM (55) or
anti-CD40 (3, 20, 21, 39, 56, 57) is widely established. mIL-4 also
enhances mouse B cell proliferation and survival (55, 58). The results
in Fig. 9
also show that IL-4 induced additional divisions of mouse B
cells stimulated with anti-CD40 Abs. Under similar conditions,
IL-13 did not significantly induce proliferation of mouse B cells
beyond that induced by anti-CD40 alone. In contrast, mIL-13
enhanced proliferation of the B9 mouse plasmacytoma cell line (5, 59),
suggesting that IL-13 might enhance normal B cell proliferation under
other circumstances. The effects of IL-13 on mouse B cells were
consistent with the known effects of hIL-13 in providing survival
signals to anti-CD40-stimulated human peripheral blood B cells
(60), B-chronic lymphocytic leukemia cells (61), and non-Hodgkins
lymphoma B cells (62). Similar to IL-13, IL-4 prolongs the viability of
not only human and mouse B cells (58, 63, 64, 65, 66) but also malignant human
B cells (67, 68).
Interestingly, in some experiments, high concentrations of anti-CD40 Ab induced purified B cells to produce Abs in the absence of any exogenous cytokine. This effect could not be inhibited by anti-IL-4 mAb, indicating an IL-4-independent stimulation pathway. Furthermore, in the presence of IL-4, anti-CD40-stimulated B cells formed tight clusters, in sharp contrast to anti-CD40 plus IL-13 cultures. As the EBV-transformed human B cell lines express IL-13 mRNA and produce a small amount of IL-13 (69, 70, 71), it is conceivable that mouse splenic B cells can produce IL-13 upon strong anti-CD40 activation and thus function in an autocrine fashion. This would be consistent with the inability of exogenous IL-13 to further enhance Ab production at high anti-CD40 concentrations.
In addition to the direct effect of IL-13 on B cells that we have demonstrated, other factors may also be involved in up-regulating Ab production during in vivo IL-13 administration and CRBCs immunization. IL-13 induces extramedullary hemopoiesis and monocytosis in mice (6). It is conceivable that the increased numbers of macrophages may secrete soluble mediators that enhance the Ab response, or that enhanced Ag presentation to T cells may provide more B cell help. This may also explain the isotype differences between the enhancement mainly of IgM levels in vitro compared with the enhancement of IgG levels in vivo. Unlike IL-4, which plays an important role in T cell differentiation (27, 72, 73, 74), IL-13 does not appear to bind to or exhibit such biological function on either human or mouse T cells (5, 7, 69). Thus, IL-13 is unlikely to affect B cells by directly altering T cell differentiation in vivo.
In comparison to IL-4, our results suggest that IL-13 may play a more
minor role during Ab responses in vivo. However, there is potential for
a more substantial role in circumstances where other B cell stimulatory
cytokines, such as IL-4, are absent or ineffective. The IL-4R
-chain
is also a component of some IL-13 receptors (8, 9, 14, 75, 76, 77, 78, 79). IL-4R
-chain-deficient mice fail to produce Ag specific IgG1 responses
(80), whereas IL-4-deficient mice generate lower Ag specific IgG1
levels (23, 81), suggesting a possible role for IL-13 in Ab production
in vivo. In addition, IL-13 is not produced in complete concordance
with IL-4, and so there may be circumstances in which IL-13 but not
IL-4 is expressed. hIL-13 is expressed within 2 h of activation
and hIL-13 mRNA can still be detected after 72 h. In contrast to
the early, yet sustained hIL-13 mRNA expression, hIL-4 mRNA expression
in T cells is transient and can only be detected 24 h after
activation (5). If mIL-13 has similar expression kinetics, it may play
an important role in stimulating an initial Ab response before the
expression of IL-4, or sustaining Ab production at later times.
| Acknowledgments |
|---|
| Footnotes |
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2 Address correspondence and reprint requests to Dr. Timothy R. Mosmann, Center for Vaccine Biology and Immunology, Institute for Biomedical Sciences, University of Rochester Medical Center, 601 Elmwood Ave., Box 609, Rochester, NY 14642. E-mail address: ![]()
3 Abbreviations used in this paper: h, human; m, mouse; CD40L, CD40 ligand; ELISPOT, enzyme-linked immunospot; HSLAS, Health Sciences Laboratory Animal Services; CRBCs, chicken RBCs; PFA, paraformaldehyde; IMDM, Iscoves modified Dulbeccos medium; CFSE, 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester. ![]()
Received for publication January 15, 1998. Accepted for publication September 2, 1998.
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mRNA expressed in human B, T, and endothelial cells encoding an alternate type-II interleukin-4/interleukin-13 receptor. Eur. J. Immunol. 27:971.[Medline]
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or IL-10. J. Immunol. 151:6370.[Abstract]
and B cell stimulatory factor-1 reciprocally regulate Ig isotype production. Science 236:944.
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A. Tomkinson, C. Duez, G. Cieslewicz, J. C. Pratt, A. Joetham, M.-C. Shanafelt, R. Gundel, and E. W. Gelfand A Murine IL-4 Receptor Antagonist That Inhibits IL-4- and IL-13-Induced Responses Prevents Antigen-Induced Airway Eosinophilia and Airway Hyperresponsiveness J. Immunol., May 1, 2001; 166(9): 5792 - 5800. [Abstract] [Full Text] [PDF] |
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R. P. Andrews, L. R. Rosa, M. O. Daines, and G. K. Khurana Hershey Reconstitution of a Functional Human Type II IL-4/IL-13 Receptor in Mouse B Cells: Demonstration of Species Specificity J. Immunol., February 1, 2001; 166(3): 1716 - 1722. [Abstract] [Full Text] [PDF] |
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H. N. Ehigiator, A. W. Stadnyk, and T. D. G. Lee Extract of Nippostrongylus brasiliensis Stimulates Polyclonal Type-2 Immunoglobulin Response by Inducing De Novo Class Switch Infect. Immun., September 1, 2000; 68(9): 4913 - 4922. [Abstract] [Full Text] [PDF] |
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S. L. Constant, C. Dong, D. D. Yang, M. Wysk, R. J. Davis, and R. A. Flavell JNK1 Is Required for T Cell-Mediated Immunity Against Leishmania major Infection J. Immunol., September 1, 2000; 165(5): 2671 - 2676. [Abstract] [Full Text] [PDF] |
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N. M. O'Brien-Simpson, R. A. Paolini, and E. C. Reynolds RgpA-Kgp Peptide-Based Immunogens Provide Protection against Porphyromonas gingivalis Challenge in a Murine Lesion Model Infect. Immun., July 1, 2000; 68(7): 4055 - 4063. [Abstract] [Full Text] [PDF] |
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B. Johansson, S. Ingvarsson, P. Bjorck, and C. A. K. Borrebaeck Human Interdigitating Dendritic Cells Induce Isotype Switching and IL-13-Dependent IgM Production in CD40-Activated Naive B Cells J. Immunol., February 15, 2000; 164(4): 1847 - 1854. [Abstract] [Full Text] [PDF] |
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